oa_GD.
'"
~
OI2..Q:O,n S'dSTCl'V\
he ce1~is the basic morphological and
T functional unit of all living things. It is
the smallest entity that has the capacity
to perform all oflife's functions and is, therefore,
capable, under favorable environmental
conditions, of independent existence. Indeed,
numerous species of unicellular organisms exist.
In fact, some, like the disease-causing bacteria
(germs) and parasites such as Entamoeba,
hisiolytica; which causes amebiasisj are of great
concern to medical science. However, as a rule,
macroscopic organisms including humans are
multicellular.
A human being actually starts as a single
cell, the fe-rtiliz~eil ovum (zrgote~, which
results from the union of the male gamete
(sp--:-ermcellj spe-rm_atozoon~ and the female
gamete (oY'um). Multiplication t~itosis~ of
the fertilized ovum and differentiation of its
progenies eventually give rise to an extremely
complex organism that consists of a staggering
number of cells (estimated at about 1014 or 100
trillion, in adults) which can be classified into
more than 250 cell types based on morphology
and function.
To erisure the human being's survival, the
cells that comprise the human body form a
tightly knit and highly organized society. Thus,
cells that perform the same general functions are
linked together by some amount of intercellular
material and/or cell-to-cell junctions to form
tissues. Tissues, in turn, bond together in
varying proportions to form more complex '
functional structures called o:rgan~ and organs
that have interrelated functions group together
to form organ s-y-stems.
The cells of the body vary in size, shape and
form not only among the different cell types
but also among cells of the same type. These
variations are largely dictated by the functions
of the cells and/or their state of activity. Despite
their wide morphological variations however,
human cells-like all eukaTxotic cells-share
a common basic structure. They consist of
a mass of cytnplasm that is encased by a ce I
membrane, and a nucleus, a structure that is
encased by-a rnrclear ~n:£elopeand embedded
in the cytoplasm.
CELL ME(~BRANI:
(ViEMBRANE; Pl"ASiVIALEfVUViA)
The cell membrane does not merely serve
as an envelope that delimits the cell from its
surroundings. It has many other functions. It
protects the cell. It determines which substances
can move in and out of the cell and regulates
the movement of these substances to and from
the cell. It provides attachment for the skeleton:
of the cell (cy:toskeletonY. It receives and-
 Fig. 1-1.Cell. The composite illustration showsthe
different components of a cell including the cell
membrane (em),mitochondria (mi), microvilli (mv),
smooth endoplasmic reticulum (sER), ribosomes
(ri), rough endoplasmic reticulu'm (rER), nucleus
(ne), nucleolus (nu), chromatin (ch), peroxisome
(pe), Golgi complex (Gel, Iysosomes (Iy), and
centrosome (ee) which cont~ins two centrioles.
Fig. 1-2. Variations in Morphology of Cells.
Photomicrographic collage illustrates the
differences in shape, and the location and
morphology of nucleus among cells. A = neuron;
B = neutrophil; C = hepatocytes; D = oocyte; E
= skeletal muscle cells. Note: The cells are not
scaled to size.

Phospholipid Molecules in the

Cell Membrane
sends out stimuli. It provides binding sites and
receptors for enzymes and other substances. It .. Each of the phospholipid molecules in the
allowsfor cell-to-cellrecognition. And, in many cell membrane has three parts: a head and two
. cell types, it forms specialized junctions with ,tails (Fig. 1-3).The head is globular, polar and
the cell membrane of adjacent cells or with the hydrophilic while the two tails are slender,non-
extracellular matrix. polar and hydrophobic. The head consists of
gtY:1:e-r..ol
that is conjugated to a nitrogenous
The cellmembrane is merely 8-10 nm thic
compound by a phosphate bridge. One of the
and is thus too thin to be distinguished-under
two tails consists of a straight-chain saturated
the light mi~osco~e tJ:::M)whose resolving
fa ¥ acHl while the other consists of an
power is onlyO.2llm (i.e.,2 millionth ofa meter)
unsaturatea fatty acid-it has a slight kink,
at best. However, in electron micrographs
but is otherwise also straight. The fatty acids-
taken with the use of the transmission (electron
of the tails are linked by covalent bonds to the
mKLus..cope('EM), the cell membrane appears glycerolof the head.
as a trilaminar structure consisting of two
electron-dense sheets that sandwich a thin In the cell membrane, the phospholipid
molecules are organized to form two closely
electron-lucent layer. Incidentally, the EM
apposed layers. In each layer, the phospholipid
can resolve objects that are 0.2 nm (i.e., two
molecules are so arranged such that the tails
billionth of a meter) apart.
occupy the inner region while the heads occupy
The cell membrane is made up mainly of the outer region (Fig. 1-3). Thus, the electron,.
~hospholip'id and R olein molecules, but it lucent iddle zone of the cell membrane as
also contains sholestero and polysacdrarides seen in the EM corresponds to the tails of the
mostly in the form of gl:y:colipias and phospholipid molecules that are bound together
by very weak intermolecular forces while the

ESTE3/\1~& GDNZ/\LES' TEXTBOOK OF HISTOLOGY

two electron-dense layers that sandwich the
electron-lucent zone correspond to the heads.
The lipid bilayer formed by the phospholipid
molecules is a highly impermeable structure
that does not allow molecules aside from
water, gases, and a few non-polar molecules
to pass freely through it. Consequently, it acts
as a barrier that regulates the entry and exit of
substances to and from the cell.
Protein Molecules
in the Cell Membrane
Protein molecules account for about
half of the mass of the cell membrane. Some,
the integra_ proteins (integral membrane
proteins, transmembrane proteins), span the
whole thickness and project out ofboth surfaces
of the cell membrane while others, called the
peri~heral roteins (peripheral membrane
proteins), are simply inserted into, or are
loosely bound to, the outer or inner surface of
the membrane.
Membrane proteins perform various
functions, including the transport of certain
substances across the cell membrane.
Cholesterol Molecules
in the Cell Membrane
Cholesterol molecules, which in the
cell membrane vary in number from a few to
almost as many as the phospholipid molecules,
are found in the irregular spaces between the
phospholipid molecules. They serve to stiffen'
and strengthen the cell membrane. They also
make the cell membrane less permeable to.
water-soluble substances.
Glycocalyx
In most cells, glycolipid and glycoprotein
molecules project from the outer surface of the
cell membrane to form a coating for the cell
called glycocalyx (eel coat) The glycocalyxis
seen in electron micrographs as a thin layer (2-
20 nm) of amorphous, electron-dense material.
Fig. 1-3.Cell Membrane. The drawing shows the
components of the cell membrane: phospholipid
molecules (pi) with their heads and tails,
transmembrane proteins (tmp), peripheral proteins
(pp), cholesterol molecules (ch), and glycolipids
and glycoproteins (gg).
It is involved in cell-to-cell recognition, cell-to-
cell adhesion, and immunological response.
Unit Membrane
Within the cell, there are many structures
that are made up of-or are enveloped by-
membranes that are morphologically identical
to the cell membrane. For this reason, the
trilaminar entity (as seen in the EM) that
comprises the cell membrane and makes up or
envelops all the other membrane-containing
structures in the ceiIis generically referred to as
the untt membrane" The unit membrane varies
in thickness from structure to structure and
even in the same structure, depending largely
I
on the amount of protein molecules it contains.
Specialized Junctions Formed,by
the Cell Membrane
(junctional complexes; intercellular
junctions; cell-to-cell attachments;
cell-to-cell junctions)
Many cells form specialized junctional
structures in localized regions of their cell
membrane that are in contact with other cells
CELL _
 p.v~e~l~e -lIf,VlCi-1Dv1

• cOOUl:C\ o.D·~QRer> S;
• 1·~~s'r.Yrb
s.o~e:,: fa r(3sc;~l
. ,q.o H~C<evlS).
,,:,~.
• '~eWlI ~re~wrosQ~~, '

Fig. 1-4. Electron Micrograph (EM) of a Cell. The image shows rough ER (rER), cell membrane or
plasmalemma (p), mitochondria- (m), centrioles (c), Golgi complex (Gc), nucleus (n), and chromatin
material (ch). x11,600 .

or the extracellular matrix. These specialized
junctions enable the cells to adhere to one'
another or with the extracellular matrix, or
communicate with each other.
Junctions that tightly bind cells t9. each
other or to extracellular matrix are classified into
occluding and adhesive. There is only one form
of occluding junction, the zonula occluilens
(tight junction; dosing belt), but there are
several types of adhesive junctions: zonula
adherens (adherens junction; adhering belt;
belt desmosome; band desmosome), fascia
adhe-rens, desmosome (macula adherens;
spot desmosome), and hemidesmosome.
Junctions that enable the cells to
communicate with each other (communicating
junctions) are exemplified by gap junctirrns
(nexus; communicating junctions) and
chemical synapses.
The structure of the zonula occludens,
zonula adherens, desmosome, hemidesmosome,
and gap junction are discussed in the chapter
on epithelial tissue. The fascia adherens is
described in the chapter on muscle tissue while
chemical synapses are taken up in the chapter
on nervous tissue.
C~TOPL SM
Cytoplasm refers to the homogenous
substance ~c:}':toFlasmicmatrix) that fills
the space that is bounded externally by the
cell membrane, and internally by the nuclear
envelope and the various formed elements
that are embedded in the cytoplasmic matrix.
The formed elements in the cytoplasm
consist of three groups: organelles, inclusions
and fibrillar elements. Organelles are more
or less permanent structures that perform
specific functions within the cell. Inclusions,
on the other hand, are generally temporary
fixtures and are often mere accumulations of
pigment, lipid, or other substances. The fibrillar
elements, meanwhile, form the cytoskeleton or
supporting framework that maintains the shape
and internal organization of the cell. In general,
an increase in the viscosity of the cytoplasmic
matrix means an increase in the number of
fibrillar elements.

ESTEBAN& (jONZI\LES' TEXTBOOK OF HISTOLOGY

,- Fig. 1-5. Mi~ochondrion. The diagram shows the parts of a mitochondrion.
Cytoplasmic Matrix (Cytosol)
The C}~toBlasmh: atri:xi is viscid,
translucent, and colloidal in nature. It is
mainly made up of water (70% or more by
volume) where a host of inorganic ions
and organic molecules (proteins, lipids,
carbohydrates, nucleic acids, enzymes,
products of enzymatic activity, etc.)
are dissolved. Although amorphous, the
cytoplasmic matrix is an important, complex,
and dynamic constituent of the cell.Itis the site
of many essential biochemical processes and it
provides a suitable milieu for the organelles in
performing their functions.
Organelles
The cytoplasmic organelles include the
mitochondria, ribosomes, endoplasmic
reticulum, Golgi complex, lysosomes,
peroxisomes, and centrosome. Except for the
ribosomes and centrosome, all the cytoplasmic
organelles are delimited by unit membranes:
Practically all cells have organelles. The
only notable exceptions are the mature red
blood cells (RBCs) and the lens fibers that
comprise the lens of the eye, which have none.
In fact, the mature RBCs and the lens fibers are
not simply devoid of organelles, they also do
~)
not possess a nucleus. The type and number of
cytoplasmic organelles in a cell are dictated by
the cell's function and state of activity.
Mitochondria
Mito'chond-ria (singular form:
mitochondrion) are organellesthat are present,
in all cell except of course in the RBCs and
lens fibers. They are often hotdog-shaped, but
they can alter their shape and become rod-lik ,
filamentous, spherical etc. They are O.Sto 1.0
~m in diameter and up to 10 ~m long.
Under the LM, mitochondria are not visible
in H&E preparations but special staining (e.g.,
supravital staining using Janus green) and
phase contrast microscopy demonstrate their
presence. They can be distinguished and their
morphology appreciated through electron
microscopy.
Under the EM, a mitochondrion is seen
to consist of a wall that encloses a space or
cavity (intercristal sp-ace) that is filled with
amorphous substance ~mitochondrial matrix).
The mitochondrial wall is made up oftwo layers
of unit membrane. The outer membrane Oli
leaflet delimits the mitochondrion from the
cytoplasmic matrix while the inner leaflet is
infolded to form shelf-like tubular structures
CELLW

Mitochondria are the "powerhouses" 0
the ceH-they generate most of the energy that
is needed by the cell to perform its metabolic
processes. They are able to produce energy
because the mitochondrial matrix contains
most of the enzymes involved in the Krebs
,tricarboxy:lic acia cycle. These enzymes
complete the degradation of the products of
fat, carbohydrate, and protein metabolism into
carbon dioxide and water. This degradation
process, which starts in the cytoplasmic matrix
but completed in the mitochondrial matrix,
yields a lot of alk_-nosinetrtpbosp ate (ltTP)
molecules that are released.into the cytoplasmic
matrix. ATP is the principal source of energy
for the various metabolic processes of the
cell. Incidentally, cells that do not possess
mitochondria such as RBes, or those with only
a few such as'muscte cells, rely on glycolysis
(i.e.,enzymaticbreakdown ofglucoseto pyruvic
acid which also releases ATP molecules) for
their energy needs.
The matrix of the mitochondrion also
to synthesize proteins and enzymes that it'
needs in performing its function. By the way,
.Q1ifocnonar.ialDNJ\)is the onlyDNA in the cell
that is outside the nucleus.
The number of mitochondria in a cell
depends on the cell's energy requirement.
For example, the dteJlatocxt , a cell that is a
"workhorse,"has more than 2,000 mitochondria
while the small I-ymghocxte, which is an
inactive cell, has only a few.
Mitochondria are motile) They tend to
aggregate in areas within the cell where energy
ESTE3!\N & (jONZI,LlSTEXTBOOK OF HISTOLOGY
requirement is high. In the sperm cell, for
example, they are concentrated in the middle
piece of the tail where they form a sheath.
Mitochondria have limited life span but
they can replicate in a manner akin to the binary
fission of bacteria.
Interestingly, mitochondria can only
be produced-or sourced from existing ones.
Hence, the mitochondria in all cells of the body
have been derived from the mitochondria of
the female gamete .(ovum) because the male
gamete '(sperm cell) does not contribute any
cytoplasmic component to the formation of the
zygote (see fertilization, Chapter XVII).
Ribosomes
1 Ribosomes (Singular:ribosome) are minute
organelles (about 15 to 30 nm in size) that can
only be distinguished via high-magnification
electron microscopy. They are seen under the
EM as small electron-dense granules that occur
singlyor in small clusters calledeolxribosume-s
o~ polysomes. Polyribosomes are composed
of ribosomes that are connected to each other
by a fine thread of messerrge-rRNi\ {mRNlt,l,
which they are actively translating (see protein
synthesis, page 17).
4 Ribosomes and polyribosomes either
lie free in the cytoplasm (free dtiosomes1
or are attached (attaelied ribosomes) to the
su'rfacesof the membranes of the endoplasmic
reticulum (ER).
3 A ribosome is made up of two subunits
(small and large); the large subunit is about
twice the size ofthe small subunit: A ribosomal
subunit, which is otherwise referred to as
db-onucleoprotein, is in the form of a dense, . ,-
globularstructure that is composed ofa strand or
dboso a R ( A) and some associated ,,---.
p.roteins of which 80 types have already been
identified. The ribosomal subunits are produced
in the nucleu -specifically, the nucleolust=
and then exported to the cytoplasm where the
small and large subunits come together to form
ribosomes.
I
I·
2 Ribosomes and polyribosomes are present
in all cells but their number and distribution
vary depending on the cell type. Cells with
numerous ribosomes have intensely basophilic
cytoplasm. The basophilia is due to the
The endoplasmic reticulum serves as
a supporting structure for the cytoplasm,
but more importantly, it is involved in the'
production of numerous substances that are to'
be used within, or exported by, the cell.!

numerous phosphate groups in the RNA of the ..

The rER receives the proteins that are
ribosomes.
synthesized by attached ribosomes. It then
5 Ribosomes are sites for protein synthesis, processes these proteins .before passing them
and are thus abundant in cellsthat produce a lot over in the form of membrane-enclosed
of proteins. It is in the ribosomes where amino structures called transfer vesicles to the Golgi
acids are assembled into polypeptide chains. complex.
6 Proteins that are produced in free The sER, on the other hand, is the site of
ribosomes are used exclusivelywithin the cell the synthesis of phospholipids, cholesterol and
(e.g., proteins of the cytoplasmic matrix as other steroids-its membranes have numerous
well as the protein subunits of structures that attached enzymes which are thus accessible to
require constant renewal such as microtubules substrates that exist within the cytoplasm. The
and microfilaments).Proteins that areproduced sER is also involved in the transport of fatty
in attached ribosomes, on the other hand, are acids and lipids.
further processedin the ER and Goigicomplex.
When these proteins 'are finally released by the In most cell types, the rERis more extensive
Golgi complex, some of them are used within than the sER. It comprisesmuch ofthe ER while
the cell while some are exported by the cell. the sER is merely made up of a small number of
tubules with hardly any cisternae and vesicles.
Endoplasmic Reticulum (ER) However, in a few cell types such as the liver
" cell (hepatocyte), the sER is better developed
The endoptasmrc reticulum is than the rER. In hepatocytes, the sER servesas
the most extensive membranous structure the site of the detoxification of various noxious
a
in the cytoplasm. It consists of system of substances. In striated muscles, the sER is
interconnecting tubules, vesicles,and flattened modified to form the sarcoplasmic reticulum
sacs (cisternae'. The tubules, vesicles, and which stores and releases the calcium ions that
sacs are fluid-filled cavities or spaces that are are needed to initiate muscle contraction.
enclosed by unit membranes which are much
thinner than the cell membrane. The ER is a dynamic organelle. It is capable
of remodeling, disassembly,and assembly,and
An ER is present in practically all cells. It
it interacts with other organelles.
is not visible in routine histologic preparations
but it can be appreciated with the use of special
Goigi complex (Goigi apparatus; Goigi
histologic techniques (e.g., with the use of
bodW .
fluorescent dyes) and distinguished under the
electron microscope. The ~olgi complexconsists ofseverallayers
of membrane-bound, smooth-surfaced, and
flattened tubes (cisternae) that are stacked on
top of each other in a semicircular manner. The
cisternae exhibit rounded dilatations (vesicles)
on their lateral ends and their cavities are filled
with fluid.
contrast, the sER is "smooth" because it has no The Golgi complex is present-albeit in
attached ribosomes. varying degrees of development-in practically

CELL

Fig. 1-6. Negative Golgi Image. The location
of the Golgi complex in cells with intensely
basic cytoplasm-such as the monocyte in the
photomicrograph-is often noticeable as a pale
region referred to as a negative Golgi image (at
arrow), Blood smear xl/000.
all cells. Some cells even have more than one
of this organelle. Under the LM, the presence
of the Golgi complex can be appreciated as
a network of solid strands in cells that have
been impregnated with si'lver salts or osmium.
It is, however, not distinguishable in H&E
preparations, although in cells with intensely
basophilic cytoplasm, its location is often
marked by a pale region (negative Golgi image)
adjacent to the nucleus. The Golgi complex can
be distinguished with electron microsc~py.
The Golgi complex has a concave surface
(maturing face; trans face) and a convex
surface (forming face; cis face). The maturing
face is the surface of the organelle that is related
to the nucleus.
The Golgi complex processes, concentrates,
sorts, and packages the proteins that it receive
from the rER. Then, it releases the proteins
into the cytoplasmic matrix in the form of
membrane-wrapped structures called secretor:y>
iVeside-s.
Some of the proteins in the secretory
vesicles are utilized within the cell (e.g.,
incorporated into developing lysosomes or
utilized as integral membrane proteins)
while some are exported by the cell (see protein
synthesis and exocytosis).
The Golgi complex, like the ER, is a very
dynamic organelle. On its forming surface,
& (jCNZALES'
ESTEB;.\P-.J TEXTBOOK OF HISTOLOGY
membrane is continually being added by the
transfer vesicles that it receives from the rER.
On its maturing surface, on the other hand,
membrane is continually being removed as
the secretory vesicles get pinched off from the
organelle.
Lysosomes
Lxsosomes (Singular: lysosome) are
chemical-containing pouches that move about
in the cytoplasmic matrix. The pouches are
made up of unit membranes and the chemicals
they contain are' h-ydrolr-tic enz:)1:mesJ
(liyal"olases)-of which more than 40 have
already been identified, including a variety of
proteases, lipases, carbohydrases, esterases,
and nucleases. The lysosomal enzymes, as
mentioned in the preceding section, come from
the Golgi complex.
Lysosomes vary in size and shape. In
general, they are spherical or ovoid bodies whose
diameter ranges from 0.05 to 0.8 ltm. Lysosomes
are present in all cells but their numbers vary
from a few hundred to thousands from cell type
to cell type and from cell to cell within the same
cell type.
Under the LM, lysosomes cannot be
distinguished in H&E preparations. The best
way to make them distinguishable is by utilizing
histochemical methods that identify their
hydrolytic enzymes.
Lysosomes constitute an intracellular
digestive system that can degrade nearly all
organic substances found in cells.Tn fact, the
only thing that prevents them from digesting
the cytoplasmic components of their own cell is
their delimiting membrane.
. Lysosomes are the prancipal organelles
involved in the cellular processes called
heterophagy and autophagy.
Heterophagy, autophagy, and
phagocytosis
Metemphag~ refers to lysosomal digestion
within the cell of a particulate material (e.g.,
r=:
--- ------------------------
 Fig. 1-7. Phagocytosis. The schematic
diagram illustrates the series of
events that constitutes phagocytosis.
Pseudopodia (ps) are formed by the
cell when a particulate material or
phagocytic material (pm) binds with
the cell's receptors. The phagocytic
material is then brought into the cell
as a membrane-bound structure called
phagocytic vacoule (pv). Subsequently,
the phagocytic vacuole is attacked
by a lysosome (Iy) which fuses its
membrane with that of the vacoule
to form a phagolysosome (pl). In the
phagolysosome, the phagocytic material
is digested and the nutrients it contains
diffuse out of the phagolysosome for
recycling. Undigested material is kept
within the phagolysosomal membrane
to form a residual body.

-
bacteria, and dead and senescent cells) that particulate material the phagosome contains.
has been brought from the extracellular The resulting memhrarre=btnmd structure
environment into the cell by phagocytosis. thatconsists of the primary lysosome and its
Phagocytosis is not common to all cells. digested material is referred to as secondary
Cells that are capable of phagocytosis such as, I¥sosome or lIhagolxsosome.
neutrophils and macrophages are referred to The nutrients derived from the lysosomal
as p'hagocy;tes. Phagocytes possess numerous digestion of bacteria and other particulate
lysosomes. materials diffuse out of the phagolysosomal
At the start ofphagocytosis,receptors on the membrane and are recycled (i.e., used within
cell surface·of the phagocyte bind the material or outside the cell); meanwhile, undigested
to be ingested (Fig. 1-7).This binding serves as materials are kept enclosed within the
-
I trigger forthe cellmembrane ofthe phagocyte to phagolysosomal membrane and are called
form cellprotrusions (p.seudopodia) at the area resitluaillotlies.
of binding. These pseudopodia slowlyelongate Residual bodies are sometimes released by
until their ends meet at the distal pole of the the cell to the extracellular area by e~ocy-tosis
material Gpbagoq"fic material) being engulfed. (see page 20), but often, they are kept within
The membranes ofthe pseudopodia then fuse to the cytoplasm. In short-lived cells, the residual
completely envelope the phagocytic material. bodies disperse into the extracellular area
The resulting membrane-bound structure, when the cell dies. In long-lived cells, however,
called phagosome or phagocytic vacuole, is they often coalesce to form inclusions called
then pinched off from the cell membrane and lipocimmtepigments (lipoluschin pigments2
drawn into the cytoplasm...
-: (see page 12).
Inside the cell,the phagosomeisapproached :A:uLoplTIIgy,
on the other hand, refers
by a p'rimarxlxsosome (i.e., a lysosome that to digestion of unneeded or senescent cell
has not digested anything yet), which fuses its organelles. Accordingly, lysosomes are also
membrane with that of the phagosome, and numerousin cellswith highturnover oforganelles
then, with its hydrolytic enzymes, digests the such as exocrine gland cells and neurons.

CELL"
There are various autophagic processes but
very often, autophagy involves wrapping the
target cell structure with a unit membrane. The
resulting vesicle is then attacked by a primary
lysosome which fuses its membrane with the
vesicle. As in heterophagy, the digested material
that results from autophagy diffuses out of the
lysosomal membrane and is recycled.
Through autophagy, lysosomes play an
important role in the structural renewal of the
cell, which is a continuous process.
As a rule, cells utilize lysosomes for
intracellular digestion only. There are, however,
some cell types that release the hydrolytic
enzymes in their lysosomes extracellularly ..
Most noteworthy is the osteoclast, the cell
responsible for bone resorption.
The oxidases in peroxisomes are important
in the process of oxidation that results in the'
detoxification and catabolism of various-
substances taken into or produced in the cell .
including phospholipids, fatty acids, ethanol
and formaldehyde. .
Oxidation of certain substances such as
long-chain fatty acids generates hydrogen
peroxide as a by-product. Hydrogen peroxide
has some beneficial uses within the cell. It is
involved in some metabolic reactions and is
utilized by phagocytes in destroying invading
microorganisms. However, it is also a potentially
cytotoxic substance. To prevent hydrogen
peroxide from reaching cytotoxic levels, cells
use the catalase in their peroxisomes to catalyze
the conversion of hydrogen peroxide (thus, the
term peroxisome) into oxygen and water.

Peroxisomes (Microbo.dies)
Centrosome (Microtubule Organizing
, PeToxisomes (singular: peroxisome), like Center [MTOC])
lysosomes, are membrane-bound spherical
The centro-some) which is otherwise called
bodies that contain chemical . The chemicals
the Microtubule Organizing Center (MTOC),
they contain are also enzymes, but unlike
is' a dense, spherical area in the cytoplasm-
lysosomes, peroxisomes do not have hydrolases, usually located near the nucleus and many
instead, they contain oxidase~ and catalase. times surrounded by the Golgi complex-that
The enzymes of peroxisomes, unlike lysosomal is present in all cells. Typically, the centrosome
enzymes, do not come from the Golgi complex, .consists of a pair of minute, short, cylindrical
but from the cytoplasmic matrix. Hence, they bodies termed centrioles that.are surrounded by
are probably synthesized in the free ribosomes. granular structures called \~entriolarsatellites.
Peroxisomes vary in diameter from O.S to 1.2
~m, resemble lysosomes morphologically, and
are difficult to differentiate from the latter even
under the electron microscope. But they can be
distinguished from lysosomes by employing the
appropriate histochemical techniques.
Peroxisomes are present in all cells but
they are particularly numerous in cells that are
metabolically active such as hepatocytes (liver
cells).

Fig. 1-8. Centrioles. The centrosome typically

contains two centrioles-cylindrical structures that
lie perpendicular to each other. The wall of the
centriole isformed by nine groups of microtubules,
each group (called a triplet) consisting of three
microtubules.

ESTEBf.\f·-.j& ccmZJ\LLS' TEXTBOOK OF HISTOLOGY

The two centrioles, which are collectively
referred to as dipioso-me, lie perpendicular to
each other. Each centriole is 0.2 ~m in diameter
and O.Sto 0.7 ~m in length.
. The centrosome is the site where
microtubule are assembled or formed:
Microtubules are fibrillar structures that form
part of the c-}':tos e eton (see page 12). They
also comprise the wall of the centrioles" the core
(~o-ne-me) of the structures that are derived
from centrioles (i.e., cihum and flagellum) as
well as mit::o:ticspindles, which likewise arise
from the centrioles.
A centriole is seen as a tubular structure
that is made up of an electron-dense wall that
surrounds an electron-lucent (hollow) space
in EM. The centriolar wall is formed by nine
groups of microtubules (Fig. 1-8). Each group
(called a triplet) consists ofthree microtubules.
When seen in eros's section, the innermost
microtubule in each triplet is circular while
the outer two microtubules are C-shaped. The
triplets are obliquelyset,but within each triplet,
the microtubules are arranged parallel to each
other. The innermost microtubule ofeachtriplet
is connected to the outermost microtubule of
the triplet adjacent to it by a fine filament.
The centrioles are the sources of the
m-itotic spindles that 'appear during mitosis.
Prior to mitosis, the two centrioles replicate.
Centrioles do not replicate by dividing; instead,
a bud (procentriole) grows out of the lateral
surface of each centriole. The procentrioles
subsequently elongate to form daughter
centrioles that are set perpendicular to their
corresponding mother centrioles. When fully
formed, the daughter centrioles separate from
their mother centrioles and thenceforth, each
mother-and-daughter centriole makes up a
diplosome that soon acquires its own centriolar
satellites to form a centrosome.
At the start of mitosis (i.e., prophase
stage), one centrosome migrates to one pole of
the cell while the other centrosome moves to
the opposite pole of the cell. Mitotic spindles,
which are made up of microtubules and some
associated proteins, then grow out from the
two sets of centrioles in both centrosomes. The
mitotic spindlespull the sister chromatids apart
during mitosis. The mitotic spindlesbreak apart
and disappear during telophase (i.e.,last stage
of mitosis).
At completion ofmitosis, each daughter cell
receives one centrosome.
Aside from forming the mitotic spindles,
the centrioles are also the sources of the cilia
of ciliated cells and the flagellum (tail) of the
sperm cell or spermatozoon. Every cilium
and flagellum is formed by growing out of a
centriole. Even when already fully formed, a
cilium or flagellum remains attached to its
parent centriole which is often referred to as
basal body.
Inclusions
Inclusions are generally temporary and
inert structures in the cell that mayor may not
be membrane-bound. They vary in size, shape,
and content. Some are useful while others
are harmful to the body. Not all cells contain
inclusions and no cell contains all the known
inclusions. The kind and number of inclusions
a cell contains depend on its function and state
of activity.
Inclusions include fat droplets (lipid
droplets), glycogen (glycogen granules),
zymogen granules, pigment granules,
crystals, and dust particles.
Eat or liRid droI'lets, are present in many
cell types, but one cell type in particular,
the adipose cell tadipoq:t1!j fat celly, is
specialized to store lipid. In this cell, the lipid
droplets coalesce to form a Single huge blob
that can occupy more than 90% of the cell. In
the cytoplasm, fat droplets are not enclosed by
membranes-they are in direct contact with the
cytoplasmic matrix.
In routine histologic preparations, lipid is
extracted by the reagents used and the areas
previously occupied by lipid droplets are seen
as mere round, clear areas. However, in tissues
CELL'"
fixed with glutaraldehyde and osmic acid, lipid
droplets are preserved and appear as gray- or
black-staining globules.
61y:c_ogen,the storage form of carbohydrates,
is an inclusion that is present in many cells, but
it is particularly abundant in liye~ ana muscle
cells. Glycogen is not distinguishable in routine
histologic preparations but it can be appreciated
in special preparations such as those utilizing
the Periodic-Acid Schiff method in which
the glycogen granules are colored purple or granules; lipofuschin pigments, Iipofuschin
magenta. In electron micrographs, glycogen granules), as mentioned in the section on
inclusions manifest as electron-dense granules
that are not enveloped by membranes. They
occur in two sizes, large calphaj partjcles and
small fbetahladicles that are about 90 nm and • are yellowish-brown bodies that are common in
20 to 30 nm in diameter, respectively.
Many kinds o£:r-igment"Sare present in cells
-as inclusions. They are in the form of granules
(pigment granules). The more commonly
occurring ones are melanin, hemosiderin, and
lipochrome.
clanin is the pigment that accounts for
the brown to black coloration of the skin where
it is synthesized by cells called mela_nocytes.
Melanin granules are also present in' the
nerve cells of the substantia igra and locus
coeruleus in the brain and the cells of pigment
epithelium of the retin~ (l.e., innermost
histologic layer of the eye). The mechanism of
melanin formation in the substantia nigra and
locus coeruleus is not well understood yet, but
in the pigment epithelium of the retina, melanin
is evidently produced in the sER of the epithelial
cells.
Hemosiderin is a brown pigment that
is the product of the lysosomal digestion of
hemoglobin, the iron-containing pigment
ESTEBA\J 8"(;;O\JZA ..ES' TEXTBOOK OF HISTOLOGY
responsible for the color of red blood cells
(RBCs). It is seen in the form of granules in cells
such as those in the spleen that phagocytose
dying RBCs. Hemosiderin granules are
membrane-bound because they are formed
inside the lysosomes. They can easily be
distinguished from other pigments by using
stains that selectively stain iron such as Prussian
blue.
bi£ochrome Figments (lipochrome
lysosomes, are membrane-bound structures that
consist of coalesced residual bodies, which are
undigested residues oflysosomal activity. They
long-lived cells such as the muscle cells in the
myocardium and Sertoli cells in the testes.
Crystals are inclusions that are present
in only a few cell types in the body, notably
the interstitial cells (of Leydig) and Sertoli
cells of the testes. Their exact chemical
composition has not been established yet. They
are not membrane-bound, and occur free in
the cytoplasm. They are often rod-shaped but
they can occur in various other shapes. They
have no known functions. They are probably
degenerative products because they are more
common in cells of older individuals.
Dust particles are numerous, in the
cytoplasm of phagocytes fpulmonaryalveolar
macrophages; du-stcells) of the lungs. They are
particularly common among smokers and city
dwellers. They are brown to black membrane-
bound structures that contain exogenous
materials such as dust and particulate carbon.
Cytoskeleton
In addition to organelles and inclusions,
the cell's cytoplasm contains a complex
network of fibrillar elements that forms the
structural framework' or skeleton of the cell
~c·y:tos}{eleton).These fibrillar elements,
which incidentally, serve other functions aside
from comprising the cytoskeleton, can only be
demonstrated with the aid of special histologic
techniques and by electron microscopy. microfilaments, intermediate filaments are
They include microfilaments, intermediate solidin crosssection, but unlike microfilaments,
filaments, and microtubules. intermediate filaments do not undergo frequent
assembly and disassembly;
Microfilaments There are many types of intermediate
M-rcrofilaments, which are present in all filaments, but the five major types that are
cells, are solid in cross section. They vary in morphologically similar but different 'in their
length but they average5 to 7 nm in diameter. protein contents are: 1) keratin, 2) desmin,
3) vim entin, 4) neurofilament, and 5) glial
A microfilament is made up o£F-adi::n,the
filament.
frlamenrous form of adin. Actin comprises
10% to 15% of the total protein in cells but Keratin fila--.mentsare present only in
only half exists as F-actin; the rest exists as epithelial cells. They are particularly numerous
C-ac_ttn fglobula .. aetin~ the soluble form of in keratinocytes, the main cell type present in
the protein. F-actin is formed when two strands the epidermis- (i.e., outer histologic layer) of
ofglobular actin subunits (G.-actin)coil around the skin. Their biochemical composition varies
each other,much like thefibers of a rope, to formdepending on the cell type. This variability can
a filament. Microfilaments can be assembled be explained by the fact that there are several
and disassembled very fast, and, in fact, they types of keratin subunits and the biochemical
composition of a keratin filament depends upon
undergo frequent assembly and disassembly to
which, and how much ofthese, subunits combine
accommodate the changes in cell shape and cell
to form the filament. The primary function of
movement.
keratin filaments is to protect epithelial cells
Microfilaments are abundant in the from mechanical and non-mechanical stresses.
peripheral areas of most cells just beneath the Those that are attached to the inner surface of
cellmembrane. Here, they are organized to form' the cell membrane are also involved in cell-to-
an interconnecting network and are involved cell attach~ent.
in activities that occur in the cell membrane
are
such as exocytosis and endocytosis (see
characteristic of muscle cells. They are more
page 20). In the central area of the cell, where
numerous in smooth than in striated (i.e.,
they probably function simply as supportive
skeletal and cardiac) muscle cells. In smooth
elements, microfilaments are less numerous and muscle cells, they form bundles while in
are scattered in haphazard fashion. striated muscle cells, they are seen around the
Microfilaments are also associated with- myofibrtls, especially in the Z-lines (discs).
and are probably involved in moving' about- Desmin filaments help maintain muscle cell
cell organelles.They are also mainly or partially architecture and structure since they connect
responsible for the contraction of such cells or anchor many cytoplasmic components.
as em De ithelial eells and musde---cells that Vimentin filaments are present in cells
exhibit contractility to a marked degree. They that differentiate from me:s-eIlchy..mesuch as
are likewise involved in the locomotion of fihroblasts and muscle cells. They are scattered
certain cells. all over the cytoplasm of these cells. Vimentin
filaments are responsible for maintaining cell
Intermediate Filaments shape and the integrity of the cytoplasm, and
stabilizing cytoskeletal interactions. They also
support and anchor the cytoplasmic organelles.
Neurofilaments are characteristic of nerve
cells Cneurons? They are present in the cell

CELL"
 Microtubules that are attached to the
organelles playa role in the movement of these
organelles within the cytoplasm while those
that are simply scattered in the cytoplasm lend
internal support to the cell. Microtubules also
comprise the wall of centrioles} the mitotic
spindles that appear during mitosis and meiosis}
the core (axoneme) of the cilia of ciliated cells}
and the flagellum (tail) of the sperm cell
(spermatozoon).

Fig. 1-9. Microtubule. In cross section, a microtubule

is seen to consist of 13 tubulin molecules that
Like rnicrofilaments, microtubules can be
surround a lumen.
assembled or disassembled rapidly}as needed.
body and all the processes of neurons. In the
processes} neurofilaments run parallel to the NUCLEUS
long axis of the processes. N eurofilaments
provide internal support for the nerve cells. The nucleus} which often occupies the
central region of the cell}is the largest structure
Glial filaments (glial fibrillary acidic
(3 to 10 llm in diameter) inside the cell. It' is
protein, GFA) are present in supportive cells
commonly round or spherical} but it occurs in
called glial cells ~neuroglial celis),.of the
various other shapes and sometimes even forms
nervous system. Their main function is to
lobes as in some white blood cells (WBCs). It
provide internal support for the cells.
is present in all cells} except in the RBCs and
the lens fibers} of course. In nucleated cells}
Microtubules the nucleus is a vital structure-its removal
Microtubules are much thicker than invariably leads to death of the eel],
microfilaments or intermediate filaments.
The nucleus} which is delimited from the
They are hollow pipes that are about 2S nrn in
diameter.
The wall of the microtubule is formed
by 1 tubulin molecules that are arranged
around a lumen. Tubulin refers to a small family
of small globular proteins. The tubulins that
comprise the wall of microtubules are formed Nuclear Envelope (Nuclear
by polymerization of al~ha-tubulin and beta- membrane)
tu ulin}the two most common members of the
tubulin family. Microtubules, as discussed in Pe} Ii k e the cell
~~.:...;.;_:,,---,e,-,-n;..-v-.;e,-l_o
the section on the centrosome} are assembled membrane} is too thin to be appreciated under
in the centrosome (Microtubule Organizing the optical microscope} but it is visible in EM.
Center or MTOC). It consists of two unit membranes Eoutercan.

ESTEBN-J& (3()\!ZN._[:';' TEXTBOOK OF HISTOLOGY


Fig. 1-10. Nucleus. The illustration shows the
components of the nucleus and its nucleolus
including heterochromatin (he), euchromatin (ee),
nucleolar organizing region (no), pars fibrosa
(pf), pars granulosa (pg), nuclear pore (np), inner
nuclear membrane (inm), outer nuclear membrane
(onm), ribosomes (ri), and rough endoplasmic
reticulum (rER).
The nuclear envelope can be regarded as a
specialized portion of the rough endoplasmic
reticulum (rER) because the perinuclear space
is continuous with the cavity of the rER while
the outer nuclear membrane is continuous with
the membranes ofthe rER, and, like much ofthe
latter, is studded with ribosomes.
Closely associated with the inner surface
of the inner nuclear membrane is a fibrillar
protein layer called fibrous l..mina (nuclear
lamina). The fibrous lamina varies in thickness,
from 30 to 100 nm, depending on cell type. It
consists offine protein filaments that are bound
to integral membrane proteins. Attached to the
fibrous lamina are clumps of cliromatin. These
chromatin clumps are visible as basophilic
areas and serve to outline the inner surface
of the nuclear envelope in routine histologic
preparations.
The nuclear envelope is perforated by
circular openings ~ uclearpores) that are 70
to 7Snm in diameter. In the pores, the inner and
outer nuclear membranes are continuous with
each other. The nuclear pores vary in number
from a few hundred to' thousands depending
on cell type. They are stabilized by the fibrous
lamina.
Each nuclear pore is ringed by a nude ..r
pore cmnplex that is made up of electron-dense
protein molecules, Some of the proteins in the
nuclear pore complex form a thin film, called
nore diap'hragm,which coversthe nuclear pore.
The nuclear pores provide a channel for the
exchange of substances between the cytoplasm
and the nucleus.
Chromatin (Chromatin material;
chromatin threads)
Chromatin refers to the 'chromosomes at
interphase.
The nucleus of all nucleated human cells
At interphase, the chromosomes are
stretched out into fine entangling threads that
are collectively called chromatin. Chromatin
forms two distinct dispersal patterns: extended
areas and condensed areas (Fig. 1-10). The
extended areas are referred to as uchromatin.
CELL.-
They are composed of portions of chromosomes
that are in the process of producing ribonucleic
acids (RNAs), specifically me-ssenger RN:A:s
EmRN:A:s) and transfer RNlts (fRN:A:s).
They do not take up stains. The condensed
areas, on the other hand, are referred to as
lieterocnromafin. They are made up of parts
of chromosomes that are not actively producing
ribonucleic acid (RNA). In routine histologic
preparations, they take up stains and form
clumps or granules (chromatin granules).
At the start of cell division or mitosis, the
chromosomes condense heavily and are seen
under the LM as basophilic, rod-like structures
that are 0.5-0.8 ~m in diameter and 3-6 ~m
in length. The chromosomes remain visible
as distinct entities while th~ cell is actively
dividing. At the end of mitosis, the chromosomes
stretch out again into fine entangling threads
(chromatin).
Human Genome
Kuman genome refers to the total
amount of DNA that is present in a human
cell. Strictly speaking, it consists of a large
diromosom_al geno-me, which refers.to the
DNA in the chromosomes and a much smaller
mitocliondrial genome) which refers to the
DNA in the mitochondria (the. only cellular
structures aside from the nucleus that contain
DNA). However, very often, people refer only to
chromosomal genome when they use the term
human genome.
The human genome is a huge database that
contains all the necessary instructions for the
synthesis of all the proteins and nucleic acids
needed by the body. The instructions are in
coded form within the DNA molecules.
DNA is a very long molecule whose
structure resembles a twisted ladder. Two
strands that are made up of deox ribose (a
sugar) and phospllale molecules form the
sides of the ladder. At intervals, the sides of the
ESTEBAN& CONZl\LES' TEXTBOOK OF HISTOLOGY
nitrogen-containing bases consist of only four
kinds: adenine (~~, thymine (1]), cytosine
tC) and guanine (G). In forming a rung of the
DNA ladder, adenine pairs with thymine while
cytosine pairs with guanine.
In the DNA molecules that comprise the
human chromosomal genom~, there are three
(3) billion nitrogen-containing bases that are
arranged in a specific order in the chromosomes.
The segment of the DNA molecule within
a chromosome that contains the unique
sequence of base pairs (DNA sequence) for
the production of a particular protein is called a
. ene. It is estimated that humans have between
30,000-40,000 genes. In as much as there are
only 23 pairs of chromosomes, it follows that
each chromosome contains numerous genes.
The chromosomes, however, are not simply
made up of genes that are placed end to end.
In fact, only about 5% of the DNA molecules
in the chromosomes encode for genes. Some of
the DNA sequences in the chromosomes exist
for structural purposes; some are involved in
regulating the use of the genetic information
contained in the chromosomes; and some exist
for no apparent purpose.
Nuclear Matrix (Nuclear sap;
nucleoplasm)
Nuclear matri is composed of water,
proteins, metabolites, and ions. The proteins
that are present in the nuclear matrix are mostly
associated with DNA molecules.
that is anchored on the fibrous lamina. The
nuclear scaffold, whose protein composition is
not known yet, has attachments for DNA loops
during DNA replication. It also has links to
the intermediate filaments of the cytoskeleton.
The nuclear scaffold is believed to be involved
in the transcription and regulation of gene
expression.
Nucleolus which are transcribed in the euchromatin areas
of the nucleus.
The rrucleotus is a spherical, highly
The rRNA molecules that are produced in
basophilic structure that is usually eccentrically
the nucleolar organizing region form the pars
located in the nucleus. It is ot enveloped by
fibrosa.
uriitmembrane and is present only in cells that
are at interphase. It disappears during prophase A newly-formed rRNA molecule
and reappears only during late telophase in both subsequently gets linked to proteins that have
been imported from the cytoplasm to produce
mitosis and meiosis. Incidentally, the nuclear
a ribonucleoprotein or ribosomal subunit.
membrane also disappears and reappears at
Ribosomal subunits form the p-arsgranules-a,
about the same time as the nucleolus during
which is a grainy area in the nucleolus.
mitosis and meiosis.
As discussed in the section on ribosomes
Tdlere .s usually one nucleolus per nucleu
(see page 6), there are two kinds of ribosomal
but in cells that are actively synthesizing
subunits: large and small, Both find their way,
proteins, there can be more than one. In cells
albeit separately, into the cytoplasm via the
such as muscle cells that"do not synthesize- nuclear pores. In the cytoplasm, at the start of
or synthesize very littlc-s-protcins, it is absent. protein synthesis, a large subunit and a small
Likewise, it is larger (up to 1.0 urn) in cells that subunit unite to form a ribosome.
are actively producing proteins than in those
that are not. The function of the nucleolus is to
PROTEIN SYNTHESIS WITHIN
THE CELL
In electron micrographs, the nucleolus
is seen to consist of three areas: nucleolar One of the mostimportant activities of a
organizing region, pars fibrosa, and par's' cell is the production or synthesis of proteins.
granulosa. Proteins comprise many essential parts of
cells and tissues. They likewise make up the
enzymes that catalyze biochemical reactions,
and hormones and hormone-like substances
that regulate the activity of cells. Proteins are
also important in the immune response and vital
be several nucleolar organizing regions per in cell multiplication.
nucleolus. The nucleolar organizing region
A protein is a polypeptide chain, which
is the place where chromosomes which have
simply means it is a molecule that is composed
nucleolar organizers gather together. The
of smaller m~lecules called amino acids linked
term nucleolar organizer refers to a segment
together by peptide bonds to form a chain-like
of a DNA molecule that contains the sequence structure. Proteins range in sizefrom 50 to more
of bases that code for a ribosomal RNA: than 5,000 amino acid molecules. Each protein
(tRNA~. \}n. humans, fivepairs of chromosomes
has its own particular sequence of amino acids
with nucleolar organizers have already been that differs from the sequence in the other
identified. In the nucleolar organizing region,
the nucleolar organizers are transcribed and
l proteins.
There are 20 kinds of amino acids (i.e.,
rRNAs are produced.
alanine, histidine, glutamine, methionine,
Ribosomal RNA (rRNA) is one of three- etc.) that are incorporated in various
ribonucleic acids (RNAs) that are needed in combinations in the numerous proteins that
proteins synthesis. The others ar~ messengerl) are in the body. Some of these amino acids, the
RNA (mRNA) and tranyfer RNA (tR~A), non-essential amino acids, can be synthesized
>
CELL.,.

/
by the cells of the body but some, the essential In the m.RNA, the code for an amino acid
amino acids, cannot. The essential amino acids, consists of a sequence of three adjacent bases
and actually also most of the non-essential ones, called a codon. For example, the codon for
are obtained from proteins that are in the food methionine in the mRNA is AUG. Each codon
that th: individual ingests. codes for one and only one amino acid, but most
Proteins from ingested food are too big to amino acids have more than one codon.
be absorbed in the digestive tract. Hence, the Aside from rRNA and mRNA, a third
digestive system first breaks down the proteins type of RNA, called transf~r RNA (tRNA),
in ingested food into amino acid molecules. is involved in protein synthesis. Transfer
These are then absorbed into the bloodstream RNA is transcribed (in a manner akin to
and transported to the individual body cells, the transcription of mRNA) from the DNA
which re-assemble the amino acids into new molecules in the nucleus, and brought to the
polypeptide chains (i.e., new proteins)-a cytoplasm.
process called protein synthesis.
One end of a tRNA molecule has a special
In the cell, protein synthesis occurs in the site to which a specific amino acid gets attached
cytoplasm but the code (i.e., DNA sequence; before the tRNA leaves the nucleus. The other
gene) for the production of a specific protein end has three bases, which are complementary
is in a chromosome in the nucleus. Hence, in to the codon of its attached
. amino acid.
, These
producing a specific protein, the DNA sequence three bases are collectively called anticoaon.
for the protein is first transcribed (copied) from For example, if the amino acid that is attached
the DNA in the nucleus to a messenger RNA to one end of a tRNA is alanine, whose codon is
(mRNA). To do this, the double-stranded DNA GCU, then the anticodon that is in the other end
molecule unwinds and unzips a little at the point of the tRNA is CGA.
where the DNA sequence for the specific protein
" In the cytoplasm, the mRNA attracts
is located. While unzipped, this short segment
ribosomes. It is the ribosomes that translate the
of the DNA molecule acts as a template for an
mRNA code and assembles the amino acids into
mRNA.
polypeptide chains (i.e., proteins). A ribosome
The basic components of RNA are the same does this by binding to an mRNA and then
as those of DNA with two major differences. In scanning the mRNA for a codon (usually the
RNA, the sugar is ribo-seinstead 0 deoxy-rillose codon for methionine, which is often AUG)
and the base thy-mine is replaced by uracik at which it should initiate protein synthesis.
Thus, in transcribing a gene, each adenine (A) When the ribosome finds the codon, it allows
. :~ - . .~
in the unzipped DNA molecule attracts a uracil a tRNA that possesses the correct anticodon
(U) while the other DNA bases attract the same (UAC, for methionine) to enter the ribosome
partners as they do in a DNA molecule, which and pair up with the codon that is in the mRNA.
means guanine (G) attracts cytosine (C), C In so doing, it is able to position the first amino
attracts G, and T attracts A. acid (i.e., methionine) that will comprise the
When the unzipped segment of the protein. The ribosome then slides along the
DNA molecule has been fully transcribed mRNA to the next codon (e.g., GCU, a codon
or copied, the single-chained RNA molecule for alanine) and then again allows a tRNA that
that has been formed, called- pre-messenger has the correct anticodon (CGA, in the case of
RNA (pre-mRNA), detaches from the DNA. alanine) to enter and pair up with the codon in
Within the nucleus, the pre-mRNA undergoes the mRNA. The amino acids (in this example,
f~rther processing (post-transcriptional methionine and alanine) that are attached to the
modification) and becomes an mRNA, which other end of the two adjoining tRNAs are then
then goes to the cytoplasm. joined by peptide bonds after which the first

Esn:BM'~& CONz/\LES' TEXTBOOKOF HISTOLOGY

Fig. 1-11. Role of Organelles in the
Production and Transport of Secretory
Materials (Le., proteins) in the Cell.
Proteins for exports are synthesized in the
attached ribosomes (r). They then migrate
to the lumen of the rough endoplasmic
reticulum (rER)where they are processed.
Afterwards, the proteins are brought in
the form of transfer vesicles (tv) to the
Golgi complex (Gc) where they are further
processed. The ready-to-export proteins
leavethe Golgi complex inside membrane-
bound sacs called secretory vesicles (sv).
Later, the protein content of the secretory
vesicles are released to the interstitial
space through the cell membrane (cm) by
exocytosis.
tRNAmolecule is released by the mRNA. Then,
the ribosome again slides along the mRNA
molecule to the next codon and in no time, a
third amino acid is added to the polypeptide
chain. As the process goes on, the polypeptide
grows longer and continues to do so until the
ribosome reaches a codon (stop codon) that
tells it to end the process and release the protein: .
More than one ribosome can translate
an mRNA at the same time, -maki ng it
possible to produce many polypeptide chains
simultaneously from a single mRNA. As
mentioned in the section on ribosomes, an
mRNA molecule, together with the ribosomes
that are simultaneously translating it, comprises
a polr-so-me or~olyribosome.
The proteins that are produced in free
ribosomes are released to the cytoplasm for use
within the cell-some become part of the cell
membrane or other unit membranes within the
cell as integral or peripheral proteins, some are
utilized by other cell organelles, etc.
In contrast, the proteins that are produced in
attached ribosomes traverse the rERmembrane
and migrate to the lumen of the organelle. From
there, they travel to the area of the rER that
is closely associated with the Golgi complex.
While in transit within the rER, the proteins
are processed and specific chemical groups are
added. Upon reaching the vicinity of the Golgi
complex, the proteins are released by the rER
in the form of small membrane-bound vesicles
{tra_Dsfe:t:\'"esicles~that pinch off from the rER.
The transfer vesicles then travel to the
forming face of the Golgi complex where
their enveloping membrane coalesces with
the membrane of the Golgi complex. Soon
thereafter, the proteins in the transfer vesicles
are released into the lumen of the cisternae of
the Golgi complex. The proteins then move
towards the maturing face of the organelle,
but while in transit within the Golgi complex,
they are further processed (i.e., sulfated,
phosphorylated, glycosylated, etc.) and then
condensed. They are also sorted out, packaged,
and labeled so they can easily reach their
intended destinations. At the maturing face of
the Golgi complex, the proteins are released
into the cytoplasmic matrix in the form of
membrane-bound pouches known as secretoryj
vesicles that bud off from the surface of the
organelle.
Some of the proteins in the secretory vesicles
are utilized within the cell (e.g., incorporated
into the developing lysosomes or utilized as
integral membrane proteins) while some are
exported by the cell. The process by which the
contents of secretory vesicles are released (i.e.,
exported) by the cell is called exocrtosis.
CELL~
MOVEMENT OF MATERIALS formation of pseudopodia'. It simply involves
invagination of the cell membrane to enclose the
ACROSS THE CE~L MEMBRANE
fluid that needs to be ingested and then pinching
The cell membrane is a semi-permeable off of the membrane-bound pinocItic vesicle
barrier, which means that it allows certain from the inner surface of the cell membrane.
materials to enter and leave the cell while it Pinocytosis that involves intake of large
constrains others. " amounts of liquid is called macro inocr.tosis
while pinocytic intake of minute amounts of
Simple molecules cross the cell membrane
liquid is referred to as microBinoc}:tosis.
by a variety of ~eans: diffustonrpassage
through ion channels, carrier transpor t: In general, pinocytic vesicles are attacked
and, active transport with the aid of pumps. by lysosoines. Lysosomes destroy the membrane
These methods of transport are described in that envelops the pinocytic vesicle and its
physiology textbooks. Bigger substances such content is released inside the cell. In some cases
as bacteria and secretory products, on the other however, the pinocytic vesicle is not destroyed
hand, are carried across the cell membrane via by lysosomes, but is transported across the cell
two mechanisms: endocytosis and exocytosis. and then its content released at the opposite cell
surface. This bulk transfer of material across the
Endocytosis cell is called transcy:tosis.

Endocytosis refers to transport of Exocytosis

substances from the extracellular space into
the cell. If the substance being transported is
solid such as a bacterium or dust particle, the
process is called phagoeytosis, which has been
discussed in connection with lysosomes (see
page 9), but if it is liquid, the process is called
pinocy;tosis).
Pinocytosis, unlike phagocytosis, is
common to all cells. This process does not
require the presence of receptors and the
Exocytosis refers to the process of
transporting substances that are inside the cell,
mostly secretor:),:products Esecretirrns)which
are in the form of se.creto_ry: ves' cles from the
Golgi complex, across the cell membrane and
out of the cell. It involves fusion of the vesicular
membrane with the cell membrane followed by
the release of vesicular contents to the exterior
of the cell.
Exocytosis generates extra cell membrane
because after the secretory product is released,
the membrane of the secretory vesicle becomes
part of the cell membrane, but this excess cell
membrane is cast off later by invagination of
the cell membrane to form a small vesicle that is
pinched off and brought to the Golgi complex.
Exocytotic release of secretory products by
cells occurs in either one of two ways: regulated
secretion or constitutive secretion.

Fig. 1-12. Pinocytosis. Pinocytosis simply involves

the invagination of the cell membrane to enclose
the fluid that needs to be ingested to form a
pinocytic vesicle (pv). As a rule, within the cell, the
pinocytic vesicle is attacked by a lysosome (Iy) and
its content released in the cytoplasm.

EST'E8;.\N& (jmU!,LES' TEXTBOOK OF HISTOLOGY

 In re-gulateosecretion,which occurs in such
cells as pancreatic acinar cells and cells of the
major salivary glands, the secretoryvesiclesare
first stored in the apicalportion ofthe cellwhere
they accumulate and get dehydrated. In the
EM, the dehydrated secretory vesicles appear
as granules (secretory granules; zymogen
granules). The content of the granules are
released only when there is a specific signal-
-
--
-
-'
,J-
usually in the form of a neural or hormonal
stimulus-to do so.
In contrast, in constitutive secretion,
which occurs in such cells as fibroblasts
and chondroblasts, the secretory products
are released from the cells as. soon as they
are formed. Consequently, cells that secrete
'I
constitutively do not have visible secretory
granules in their cytoplasm. ..
. ,CELL"
 Epithelial Tissue
(

n the body, cells that have ;imilar or closely

I
organ where the tissue is located and on its
related functions are bound together by assigned function within the organ. Thus, basic
interceHubr subsranoe- (intercellular tissues are best regarded as broad categories that
m a t er i a I: extracellular sub s t a n ce , contain many subcategories.
extracellular matrix) and/or ceU-to-
cell junctiuns to form larger structural OF
and functional units called tissues. The
extracellular matrix that exists between cells
consists of an assortment of macromolecules
that are manufactured by the cells and exported
into the intercellular space. The kind, amount
and arrangement of macromolecules in the
extracellular matrix vary from type to type and
from subtype to subtype of tissues. are present between its cells. Furthermore, the
!~

A cursory examination of the microscopic
anatomy of the various parts of the body will
make one rightfully conclude that there .are
numerous types of tissues. However, a more
thorough study of the structure and behavior
of the various tissues will show that they can
actually be grouped into a handful of categories.
In fact, histologists generally agree that there are
only four basic tissues in the body: 1) epithelial
tissue, 2) connective tissue, 3) muscle tissue,
and 4) nervous tissue.
Each of the basic tissues has a set of
morphological and functional characteristics
that distinguishes it from the others. At the same
time, each exhibits numerous morphological
variations that depend on the function of the
cells that comprise epithelial tissue exhibit
polarity, The basal, lateral, and superficial
(apical) surfaces of an epithelial cell can be
determined by the location of its cytoplasmic
organelles and surface modifications.
Embryonic Origin of Epithelial
Tissues
Of the basic tissues, only epithelial tissue'S
are derived from all three embryonic germ
layer. Epithelia that cover the external surfaces
of the body (i.e., skin and its appendages, and
cornea of the eye) are derived from ectoder .
Those that form part of the digestive tract
(except for those in the mouth and anus, which

-I
are derived from ectoderm), liver, gallbladder,
pancreas, respiratory tract, urinary bladder
and urethra arise from endoderm. The
epithelia that are present in the heart, blood
and lymphatic vessels, and the serous cavities
together with those in the urinary system
(except for the urinary bladder and urethra),
and in the male and female reproductive
systems come from mesoderm.

Basal lamina Fig. 11-1.Basement Membrane (at tip of arrows).

The cells that comprise an epithelium are
arranged in one to several layers,but regardless
ofthe number of celllayers,the basal surfaces of
the cells in the deepest layer of any epithelium
rest on a thin sheet of amorphous extracellular
material called basal lamina. It is a mixture
of more than SO kinds of gl~proteins,
several types of collagen and a variety of
IT. oteoglIcans.
The basal lamina is usually about 20-100
nm thick, although in some places, such as the
glomerulus of the kidney, it is much thicker ..
It GEYm:0' ~listt-nguish_ed
under the LM but
in EM, it is typically seen to consists of three
layers-two electron-lucentlayersthat sandwich
an electron-dense one. The electron-lucent
layer where the basal surfaces of the epithelial
cells rest is the {amina rar externa (lamiM
lucida). It is largely made up of glycoproteins
including laminin and proteoglycans. The
electron-dense layer is called the lamina
d~nsa. It consists mainly of a network of fine
filaments consisting of cullagen t:y.:peV and
proteoglycans. The other electron-lucent layer,
which is not always present, is called lamina
nra irrterna. It is morphologically similar to
the lamina.rara externa but thinner and often
indistinct. Its main constituents are collagen
ty-p :v I, fib-ro~nectin,tliro_1l1_}jospo~ndin
and
p~oteoglJ::cans.
The basal lamina provides structural
support to the overlyingepithelium. In addition,
it serves as an impermeable barrier that allows
only water and small molecules to pass through,
Trachea, H&Ex400.
and limits the contact between epithelial cells
and the other cell types in the tissue.
Although it characterizes the epithelial
tissue, basal lamina is not exclusively seen or-
produced in this tissue. Basal lamina also
completely envelops individual muscle cells,
fat cells, Schwan cells (supporting cellsin the
peripheral nervous tissue), cells of the adrenal
meaulla and a few other special cell types.
The basal lamina in the epithelial tissue
is the product of the epithelial cells. It is not
pierced by blood vessels and since there are no
blood vessels in epithelium either, epithelial
cells rely on the diffusion of gases and nutrient'S
across the basal lamina for their sustenance.
Basement Membrane
underlying tissue. The lamina fibroreticularis
is 200 to sao nm thick and is thus much thicker
than the basal lamina. It is absent in some
epithelial tissues, notably in the glomeruli of
the kidneys and the capsule of the lens of the
ey:e.
The lamina fibroreticularis is a product of
onnective tissue cells s ecificall fib~rublasts,
and not of the epithelial cells.

EPITHELIAL TISSUE
 The basal lamina and the
fibroreticularis are collectively referred
as basement membrane a structure
enough to be appreciated under the LM as a
homogenous layer of extracellular material.
GE ER L CLASS F CAT. N
AND FUNCTION OF
EPITHELIAL TISSUES
The epithelial tissues in the body can
be categorized into two groups: 1) surface
epithelium; and, 2) glandular epithelium.
SIITfaceepithelium refers to epithelial tissue
that covers the external surfaces (covering
epitlielium) such as the skin, and lines the
internal surfaces (lining epitlielium) such as
the luminal surfaces ofvisceral organs and ducts
of glands of the body.
on the other
hand, refers to epithelial tissue whose cells
are specialized to elaborate (synthesize;
produce) and release (secrete) macromolecules
(secretions),
There is an overlap between the two
categories of epithelial tissues because some
surface epithelia are also secretory.
Surface Epithelium
The function of a surface epithelial tissu
depends on its location. For example, in the
skin, it serves a protective function; in the
gastrointestinal tract, it serves as an absorptive
surface. Furthermore, some surface epithelial
cells have special functions. For instance, the
neuroepithelial cells of the taste bud in the
tongue and the olfactory cells of the olfactory
epithelium in the nose are sensory cells; the
epithelial cells in the kidney are responsible for
excreting waste products and maintaining fluid
and electrolyte balance; and, the epithelial cells
in the testes are the sources of germ cells.
ESTEBAN & GONZALES TEXTBOOK OF HISTOLOGY
lamina
to
thick
Fig. 11-2.Types of Simple Epithelial Tissue. A)
Squamous; B) Cuboidal; C) Columnar; and D)
Pseudostratified.
Classification of Surface Epithelial
Tissues
Surface epithelia are classified into simple
and stratified on the basis of the number of
layers that the cells form. An epithelium that
consists of a single layer of cells is referred to
.as simpl while an epithelium that consists
of more than one layer of cells is referred to as
stratified. Simple and stratified epithelia are
further classified according to the shape of
the cells that comprise the epithelium and the
surface modifications that the epithelial cells
exhibit (e.g., if most of the cells forming the
epithelium are provided with cilia, then the
epithelium is called a ciliated epithelium.)
Simple Epithelial Tissues
If the cells that comprise a simple
epithelium are flattened, the epithelium is
termed s uamous; if they are equally tall as
wide, the epithelium is labeled cuboidal; and, if
they are taller than they are wide, the epithelium
is called columnar.
Simple Squamous Epithelium
Simple squamous epithelium consists of
a single layer of flattened cells whose nucleus
occupies the thickest part of the cells. In
some cells, the nucleus bulges prominently on
Fig. 11-3. Simple Squamous Epithelium. Arrows Fig. 11-4. Simple Cuboidal Epithelium. Arrows
point to the squamous epithelial cells. Kidney, point to the cuboidal epithelial cells that form the
H&E x400. lining of a duct. Parotid Gland, H&E x400.

the surface while in others, it is more or less surface of the heart, blood and lymphatic
flattened along the long axis of the cell and does vessels is called ~ndotlieltum. Incidentally,
not bulge on the surface. epithelium that 'lines the under surface of
When seen from the surface, squamous the cornea of the eye is also referred to as
.epithelial cells exhibit irregular polygonal endothelium, but the corneal endothelium
outlines that fit into each other like pieces is.a simple cuboidal, not simple squamous,
of a jigsaw puzzle. The cell outlines are best epithelium.
appreciated in specimens stained with sil:v:e~
d es because these dyes get deposited into the Simple Cuboidal Epithelium
intercellular substance between adjacent cells.
Simple cuboidal epithelium consists -of a
Simple squamous epithe'iium lines single layer of cells whose height approximates
the lung alveoli, the parietal layer of the their width. Side view of this epithelium reveals
Bowman's capsule in the kidneys, and many cells that are squarish in outline and whose
other structures. By the way, the shape of some nuclei are round and centrally situated while the
epithelial cells depends on their functional top view shows cells with polyhedral outlines
state. For example, in inactive thyroid follicles, that fit each other quite snugly.
the epithelial cells form a simple squ~mous or
cuboidal epithelium while in active follicles,they Simple cuboidal epithelium is present in
form a tall cuboidal or columnar epithelium. certain segments of the ducts of the major
Likewise, the granulosa cells of ovarian salivary glands and the pancreas, collecting
follicles form a simple squamous epithelium tubules of the kidney, some follicles of the
in primordial follicles but the cells become thyroid gland, and the surface of the ovary.
cuboidal and later the epithelium stratifies-when
the follicle transforms into a primary follicle. Simple Columnar Epithelium
Some .:simple squamous epithelia have Simple columnar epithelium consists
traditionally been assigned special names: the of a single layer of tall cells. The nuclei of the
simple squamous epithelium that lines the cells form, more or less, a single row. They
serous cavities of the body (e.g., pericardium, are generally oval, more basal than apical in
peritoneum, and pleura) is referred to a'S location, and their long axes lie parallel to those
mesothelium· and that which lines the luminal of the cells.

EPITHELIAL TISSUE __

Fig. 11-5.Simple Colprnnar Epithelium (Ciliated).
Arrows point to the cilia on the surfaces of the
epithelial cells. Gallbladder, H&E x400.
oviducts.
Pseudostratified Columnar Epithelium
is a variant of simple columnar epithelium
that exists in certain parts of the body.
Pseudo stratified epithelium literally means
"false stratified epithelium." It consists only
of a single layer of tall or columnar cells, but on
hurried examination under the LM, one may
be misled to think that this is a stratified type
of epithelium since the cells vary somewhat in
shape, and their nuclei are disposed in various
levels. A closer look, however, will show that the
cells constitute only one layer because they all'
rest on the basal lamina.
Pseudostratified columnar epithelium
lines the membranous and spongy parts of the
male urethra. A subtype of this epithelium (a
subtype because the cells are ciliated), referred
to as -iliat_edps_euilostra ifie columna-r:
G
epithelium or simply resptra ory:epithelium,
lines the larger passageways of the respiratory
system like the trachea and the main bronchi. Epithelium. Lip, H&E x400.
[STEB!\N & GCNz/\.LES'TEXTBOOK OF HISTOLOGY
--- ------------------------------------------------------~----------~-
Stratified Epithelial Tissues
The various stratified epithelia in the body
are classified on the basis of the shape of the
cells in their most superficial layer. Thus, if the
cells in the most superficial layer are flattened,
the epithelium is called stratified squamous;
if their height approximates their width, the
epithelium is referred to as stratified cub-oidal;
and, if they are taller than they are wide, the
epithelium is labeled stratified coltfiifiiir.
_-
Stratified Squamous Epithelium
In this type of epithelium, the cells in the
most superficial layer are flat or plaque-like,
while those in the deeper layers are tall cuboidal
or even columnar. In this epithelium, as one
examines (under the LM) the cell layers from
the deepest to the most superficial, one notes a
progressive diminution in the height of the cells.
In stratified squamous epithelium, new
cells are formed only in the deep layers. The
superficial cells are "old" cells that have been
p~~hed to the surface by the newly-formed ones,
Stratified squamous epithelium can
withstand rubbing more than any other type of
epithelium. .
When stratified squamous epithelium is
"dry,' the cells of the most superficial layer are
dead cells that have no nucleus and organelles.
This type of epithelium is termed keratini-zed.
Keratinized stratified sguamous epithelium
is impervious to water. It forms the outer
Fig. 11-6.Nonkeratinized Stratified Squamous

Fig. 11-7. Keratinized Stratified Squamous
Epithelium. Skin, H&E x400.
histologic layer of the skin, where it is referred
to as e_piderm-=is
In contrast, when the stratified squamous
epithelium is "wet," (i.e., kept moist by glandular
secretions), the cells in the most superficial layer
are flattened but still nucleated. This type of
epithelium is called emkeratinized, a misnomer
because the cells of the epithelium, like those in
keratinized stratified squamous epithelium, also
contain keratin. Nonkeratinized stratified
~guamous ep'ithelium lines the oral cavity,
esophagus, vagina, part of the urethra, and the
-
... n:ost superficial layer of the cornea of the eye.
Fig. 11-8. Stratified Cuboidal Epithelium.
The photomfcrograph shows the two layers
of cuboidal cells that line an interlobular
duct. Parotid Gland. H&E x400.
Fig. 11-9. Stratified Columnar Epithelium.
In this epithelium-which consists of two
layers of cells-the cells in the superficial
layer are columnar but the cells inthe deep
layer are cuboidal. Parotid gland, x400.
Stratified Cuboidal Epithelium
Stratified cuboidal epithelium usually
consists of two or three layers of cuboidal cells.
It is the epithelium that often Iines the larger
ducts of some glands such as the major salivary
glands. .
Stratified Columnar Epithelium.
Stratified colu~nar epithelium consists of
at least two layers of columnar cells although
sometimes the cells of the deeper layer/s are
cuboidal. It lines the large ducts of some glands.
Transitional Epithelium
There is a e of stratified epithelium,
unique to mammals, called transitional
~-ithellum that manifests features that are
in between stratified squamous and stratified
cuboidal epithelia.
Transitional epithelium lines the urinary
passages (i.e., renal calyces, renal pelvis, and
ureter) and the urinary bladder. This type of
epithelium is designed to withstand stretching.
It is essentially a stratified epithelium that is
intermediate between stratified cuboidal and
stratified squamous epithelia .
EPITHELIAL TISSUE va
 Fig. 11-10.Transitional Epithelium. The
umbrella cells that characterize this
type of epithelium are indicated by the
arrow. Urinary bladder, H&E x400.

In the contracted state of the urinary in cells where they are particularly numerous
bladder and urinary passages, the basal cells of, such as those that line the small intestine, they
the epithelium are cuboidal while the superficial form a fuzzy, fine vertical line on the surface of
cells (referred to as "umb~e-lla cells"-~ bulge the epithelium called triated or(ier or brush
out into the lumen giving the cells a dome- border. IT'ypically,microvilli are coated on their
shaped profile. In the distended bladder and outer surface by glycocalyx.
the urinary passages, however, the superficial
cells get stretched and flatten out, effectively
transforming the epithelium into a thin
stratified squamous epithelium.
- --
Surface Modifications
of Epithelial Cells
Most epithelial cells exhibit modifications
on their apical (superficial), lateral and/or basal
surface/s that are necessary for the discharge of
their specific functions. Fig. 11-11.Microvilli. Arrows point to the microvilli
on the surface of epithelial cells. Jejunum, H&E
x400.
Modifications on the Apical Surfaces
of Epithelial Cells The core of a microvillus is formed by a
network of actin fHamen .s, that is attached to
The specialized structures on the apical
the inner surface of the plasma membrane.
surface of epithelial cells include the microvilli,
cilia, flagella, and stereocilia. Microvilli number from a few to several
thousands per cell depending on the cell type.
Microvilli Their function is to increase the surface area
of the epithelium-by as much as thirtyfold, in
Many epithelial cells have short (about
some organs.
1.0 ~m long) and fine (about 0.08 ~m in
diameter) finger-like extensions or processes
Cilia (Kinocilia)
of the plasma membrane called microvilli
(singular: microvillus) that protrude from or: kino cilia (Singular: cilium;
the apical surface of the cells. Microvilli are not kinocilium) are present in the apical surfaces
individually distinguishable under the LM, but of cells that are specialized for transport of fluid

[STEl\!\f\] & CiONZf\l.ES' TEXTBOOK OF HISTOLOGY

 Stereocilia
"0°000 Stereocilia (Singular: stereociliuml are
"
.., 0
met') simply microvilli that are as long as cilia. Like
microvilli, they are non-motile and their core
00 em
consists of actin filaments.
Stereocilia are characteristic ofthe epithelial
cells that line the ductus (vas) epididymis and
ductus deferens, long tubes that help convey
sperm cells from the testes to the external'
Fig. 11-12.Cilium. Cross section of a cilium showing environment. They are also present on the hair
the microtubules (me)that comprise the axoneme. cells of the inner ear where they playa role in
Externally, the axoneme is enveloped by cell auditory and vestibular perception.
membrane (em).

or mucus over the surface of the epithelium.

- They are finger-like extensions of the plasma
-'
membrane that, unlike microvilli, .can be
distinguished under the LM, because at 7 to 10
~tmin length and 0.2 ~m in thickness, they are
much longer and thicker than microvilli.
As seen in the EM, the core Gaxoneme)
of a cilium consists of microtubules that run
parallel to its long axis, The axoneme (called
a 2:+=2 aro-neme) consists of a pair of centrally'
located microtubules that is surrounded by nine
pairs (doublets) of microtubules. '"
Cilia are motile. They beat sequentially OF Fig. 11-13. Stereocilia. Arrows indicate the
synchronously in one direction and are thus stereocilia on the surfaces of the epithelial cells.
helpful in propelling substances over the surface Ductus Epididymis, H&E x400.
of the epithelium.'
Modifications on the Lateral
Flagella Surfaces of Epithelial Cells
(intercellular junctions; cell-to-cell
agella (Singular: ftagellum) are simply
long cilia. In humans, ~ one cell tYEe attachments; junctional complexes)
possesses a flagellum, the s ermatozoon. The Epithelial cells form continuous sheets
sR r atozoon has only one flagellum, which is and are able to communicate with each other
otherwise called tail. because adjoining cells are attached to each
The tail of a spermatozoon is more than SO other on their lateral surfaces by membrane
~m long and its main function is to propel the and cytoskeletal specializations. The simplest
cell along the female genital tract. As likewise of these cell-to-cell attachments, which some
mentioned in connection with the centriole, a epithelial cells exhibit, consists of finger-like
flagellum, like a cilium, grows out of a centriole cytoplasmic processes that interdigitate with
(also called basal body) to which it likewise "similarprocesses of adjacent cells.
remains attached even when it is already fully In addition to interdigitation, epithelial
developed. cells form four other types of specialized

EPITHELIAL TISSUE __

Fig. 11-14.Illustration of Intercellular Junctions
of Eplthelial Cells
intercellular junctions that are distinguishable
under the electron microscope: 1) zonula
occludens, 2) zonula adherens, 3)
.desmosome, and 4) gap junction. The first
three are designed to keep adjacent cells in a
surface epithelium glued together thus ensuring
that substances can enter or leavethe underlying
tissue only by passing through, rather than
between, the surfaceepithelialcells.Meanwhile,
gap junctions are designed to enable adjacent
'cells to communicate with each other. By the
way,the zonula occludens and zonula adherens
are collectively referred to as juxtaluminal
junctional complex or terminal bar.
The four special types of junctional
complexes are all present and are well-
developed in simple cuboidal and simple
columnar epithelia such as those that line the
gastrointestinal tract. In many other epithelia,
however, not all the junctional complexes are
seen. For example, in the epidermis (i.e.,
the epithelial covering of the skin), the only
.junctional comple_xthat existsbetween the cells
is the desmosome.
Zonula Occludens (Tight Junction;
Closing Belt)
When present, the zonula occludens is
the most apically situated of the junctional
ESTEB;\\1 ,} C30f\iZ/\l_ES' TEXTBOOK OF HISTOLOGY
complexes.It is located on the lateral surface of
each epithelial cell immediately below the free
surface of the cell. It forms a band, 0.1 to 0.3
llm thick, which completely surrounds the cell.
At the zonula occludens, the cell membranes
of adjacent cells stick to each other (without
any intervening intercellular substance), and
in severalplaces, the adjoining cell membranes
actually fuse together. At the points of fusion,
the adjoining cells share a common cell
membrane, which is seen as a single trilaminar
structure in the EM.
Zonula Adherens (Adherens junction;
Adhering belt; Belt desmosome; Band
desmosome)
The zonula adherens is typically located
just below the zonula occludens. Like the latter,
it forms a band that completely encircles each
epithelial cell. At the zonula adherens, the cell
membranes of adjoining cells are very close
to each other, but they neither adhere nor
fuse. Instead, they are separated by a narrow
intercellular space that is merely IS to 20 nm
wide. This intercellular space is filled with
extracellular material that binds the apposed
cell membranes to each other.
The cytoplasmic surfaces of the apposed
membranes that form the zonula adherens
contain a bundle of fine filaments. In cells
that possess a terminal web-a layer of
microfilaments (actin filaments) and
intermediate (keratin) filaments that form
a network across the cell just below its apical
surface-these fine filaments serve as anchor
for the terminal web. A terminal web is present
in some epithelial cells, notably those with
microvilli, where it serves a supportive or
cytoskeletal function .
Desmosome (Macula adherens; Spot
desmosome)
Desmosomes are usually situated just
below the zonula adherens, but they may be in
other areas on the lateral surface of epithelial
cells. Unlike the zonula occludens and zonula
adherens, they do not form a band around the
epithelial cell. Instead, they form button-like or
rivet-like adhesions that are arranged in a line
around the cell. Under the LM, they sometimes
aJ?pear as a thickening of the cell membrane.
A desmosome consists of an ovoid protein
disc or plaque that is split into halves. Half of
the disc is firmly attached to the cytoplasmic
surface of the cell membrane of an epithelial cell
while the other half is similarly attached to the
cytoplasmic surface of the cell membrane of the
adjoining cell. Within each of the apposed cells,
.numerous intermediate (keratin) filaments
converge and insert into each half-disc.
At the desmosome, the cells are separated
by a relatively wide (about 30 nm) intercellular
space, which contains numerous transverse
filaments and adhesion proteins that bind the
apposed cell membranes to each other.
Desmosomes are particularly numerous in
the epidermis of the skin where, as previously
mentioned, they are the only type of junctional
complex present.
Gap Junction (Nexus; Communicating
junction)
Gap junctions referto broad areaswhere the
plasma membranes of adjoining epithelial cells
are closelyapposed but not fused to each other.
At the gap junctions, the cells are separated
by an intercellular space that is only about 3
nm wide and the apposed plasma membranes
are perforated by numerous tiny tubes called
connexons. Gap junctions enable adjacent cells
to exchangeions and small moleculesbecause at
the gap junctions, the connexons of adjoining
epithelial cells are aligned and continuous with
each other.
Modifications on the Basal Surfaces
of Epithelial Cells
The basal surfaces of some epithelial
cells contain specializations that serve for
better attachment or for more efficient
functioning.
One form ofbasal surfacemodificationisthe
emidesmoso e that exists in some epithelial
cells such as those in the stratum bas ale of
the epidermis of the skin. A hemidesmosome
is structurally identical to half a desmosome,
thus, the name.
Hem ide smo some s help anchor the
epithelial cells to the underlying basal lamina.
Another type of basal surface modification
comes in the form of basal infoldings of the
plasmalemma such as those that are present in
cells Iiningsome segments of the renal tubule .
Basal infoldings of the plasmalemma are seen
as basal striations under the LM.
Basal infoldings increase the absorbing
capacity of a cell.
Glandular Epithelium
The glandular epithelium in the body is
organized to form the functional component/s
or secretory unit/s of structures called glands
that produce substances (sec:retions~that are
needed by the body.
On the basis of where they release their
secretions, glands are categorized into two
groups: exocrine and endocrine. iExocrin_--e
~la ds deliver their secretions into the surface
epithelium while endocrine glands deliver
their secretions into the blood or lymph.
Exocrine glands whose secretory units
are located some distance from the epithelial
surface transport their secretions to the
epithelial surface by wayoftubular passageways
called ducts. Endocrine glands, on the other
hand, are ductless.
Endocrine Glands,
Mos( of (h,e endoic~ne glands arise in
the embryo as invagination or evagination
of thecovering .epithelium of body .cavities.
The invaginated or evaginated epithelium
subsequently acquires supporting structures-
connective tissue, blood vessels, and nerves-
and develops into a gland.
EPITHELIAL TISSUE ..
 In most instances) a developing endocrine
gland remains initially connected to its site of
origin by a duct) but later this connection is
severed and the gland becomes ductless. Some
endocrine glands) however) arise by migration
of epithelial cells without ever forming a duct.
The secretions that endocrine glands
elaborate are called hormones. A hormone is a
chemical substance (steroid) peptide) or amine)
that is carried by blood to organs or tissues
that have cells (target cells) which contain
appropriate receptors for it. It acts as a chemical
messenger that enables an endocrine gland to
exert its influence on its target cells) tissues) and
organs.
Endocrine glands exist as distinct organs
such as the adrenals and thyroid gland) or
as components of organs such as the islets
of Langerhans in the pancreas. They are
discussed in the chapter on the endocrine
system.
Exocrine Glands
..
Exocrine gla.nds are classified according
to the number of cells that comprise them into
unicellular or multicellular glands.
Unicellular Glands
Fig. 11-15. Goblet Cell. The goblet cell in the
diagram is indicated by the arrow.
ESTE3f.\f;j & GC)NZ!\LES' TEXTBOOK OF HISTOLOGY
Fig. 11-16.Goblet Cells. Arrows point to goblet _-
cells that form part of the surface epithelium of an
intestinal villus. Jejunum, H&E x400.
The goblet cell is one of the cell types
that constitute the surface and glandular
epithelium of many segments of the digestive
and respiratory tracts: It is a columnar cell
that is cup-shaped ~hen seen in routine
histologic preparations. Its tapered base rests
on the basal lamina while its apical portion)
called theca) is expanded to accommodate
numerous membrane-bound secretory vesicles
that contain mucin) a glycoprotein which when
mixed with water forms mucus: The secretory
vesicles push the nucleus and most of the
cytoplasmic organellesofthe goblet celltowards
the basal surface of the cell. They do not take up
the dye well and they coalesce during routine
histologicpreparation)thus the area they occupy
is simply seen as a pale-staining region in these
preparations.
Multicellular Glands
There are three categories of multicellular
glands (i.e., glands that consist of more than
one cell): 1) secretory epithelial sheet) 2)
intraepithelial gland) and 3) glands with
ducts.
Secretory Epithelial Sheet
ecretor¥ epifhelial sheet refers to any
surface epithelium) where most) if not all)of the
cells are secretory. An example of a secretory
epithelial sheet is the ependyma) the simple
cuboidal epithelium that lines the choroid
Fig. 11-17 (above). Secretory Epithelial
Sheet. This type of multicellular gland is
made up of a single layer of epithelial cells
where all the cells are secretory.

Fig. 11-18(right). Secretory Epithelial Sheet.

The single layer of cuboidal cells indicated
by the arrows comprise a secretory epithelial
sheet. Choroid Plexus, H&Ex400.

plexuses in the brain, and which produces
cerebrospinal fluid (CSF).
Intraepithelial Gland
Irrtrae£ithelial gland. refers to a group of
secretory cells in a surface epithelium that gather
together around a small orifice-that serves
as a duct-and form shallow invaginations
~ithin the epithelial surface. Examples of
intraepithelial glands exist in the epithelium
that lines the penile urethra.
Exocrine Glands with Ducts
Exocrine glands with ducts consist of glands
that possess "true" ducts. They are the most
complex of the exocrine glands. Their secretory
units, which lie underneath the epithelium, are
connected to the epithelial surface to which they
deliver their secretions by a duct or a system of
ducts.
Exocrine glands with ducts arise as
invaginations of surface epithelia. Unlike
endocrine glands that arise similarly but which
invariably lose their ducts, exocrine glands
retain their tubular connections (ducts) with
the surface epithelium even when they are
already fully formed.
Exocrine glands with ducts range in size
from microscopic structures to large distinct
organs such as the major salivary glands,
pancreas, and liver.
Classification of Exocrine Glands with
Ducts According to Morphology
Exocrine glands with ducts are classified
and sub-classified according to: a) complexity
of their duct/s; and b) morphology of their
secretory units (l.e., the cluster of cells that
produce secretion or secretory material).
If a gland has a single unbranched duct,
it is called a simp-Iegland, but if the duct has
branches, it is referred to as a comBound gland.
The duct system of large compound exocrine
glands, such as the major salivary glands, is
discussed in the chapter on the accessory
organs of the digestive system.
The secretory units of exocrine glands
are in the form of either blind-ending tubes
Fig: 11-19.Types of Simple Exocrine Glands with
Ducts. A) Simple tubular; B) Simple coiled tubular;
C) Simple branched tubular; D) Simple alveolar
(acinar); E) Simple branched alveolar (acinar). The
secretory cells are red while the ductal cells are
green.

EPITHELIAL TISSUE ..

(secretory tubules), or globular or basket-like
structures (acini; alveoli). If all its secretory
units are in the form of secretory tubules, the
gland is a tubular glana; if all its secretory
units are in the form of acini (singular: acinus1
or alveoli'(singular: alveoluSi),the gland is an
lveo ~ or acimnrs gland; and, if some of
the secretory portions are tubular and some
are globular, the gland is a tubuloalveolar
gland ftubuloacinous glana). Tubular and
acinous glands are further classifiedinto either
branched or coiled when their secretory units
are branched or coiled!respectively.
Examples of simple tubular glands are the
crypts of Lieberkuhn (intestinal glands) in the
intestines. The cardiac glands in the stomach,
on the other hand, exemplifysimple branched
tubular glands while the sweat glands and
sebaceous glands in the skin illustrate simple
coiled tubular and simple branched alveolar
glands.fespectively, The Brunner's glands in
the duodenum (i.e., first segment of the small
intestine) typify compound coiled tubular
glands while the major salivary glands (i.e.,
parotid, submandibular, and sublingual) are
examplesofcompound tubuloalveolar glands.
Classification of Secreto-ry Cells, Acini and
Exocrine Glands According to Nature of
Secretion
The cells that comprise the secretory
units (tubules or alveoli) of exocrine glands
are classified into two types according to the
ESTEBAN& GONZALES'TEXTBOOK OF HISTOLOGY
Fig. 11-20.Types of Compound Exocrine
Glands. A} Compound tubular; B}
Compound alveolar (acinar);C) Compound
tubuloalveolar (tubuloacinar). The
secretory cells are red while the ductal
cells are green.
nature of their secretion: 1) mucous (mucus-
secreting); and, 2) serous (serous-secreting).
MuclJUScells produce a viscous secretion
that contains mucin, a substance rich in
glycoproteins that when hydrated, forms
mucus that protects and lubricates covering
epithelia.Their nucleus is highly condensed and
flattened, and is pushed towards the basal area
by secretory granules that occupy most of the
cell. Their secretory granules, however, do not
take up the dyes well and, in routine histologic
preparations, are poorly preserved. Hence, in
routine histologic preparations, mucous cells
have pale-staining cytoplasm.
Serous cells, on the other hand, produce
a thin, watery secretion that often contains
enzymes. They are columnar cells with
spherical and basally located nucleus. Their
cytoplasm is basophilic especially around the
nucleus. Their secretory granules, which take ..
up the dyes rather well,are mostly located in the
apicalregion of the cell.
Secretory units of exocrine glands that
consist entirely of mucous cells are called
mucous alveoli tacini})while those that consist
exclusively of serous cells are called serous
a eoli (acini). Those that contain both mucous,
and serouscellsare calledmixed lveoli ~_cjnJJ.
In some mixed alveoli where most of the
secretory cells are mucous cells, such as those
found in the submandibular gland, the few
serous cells that are present sometimes form
 Fig. 11-21(left). Simple Tubular Glands. The arrow
points to the orifice of one of the several simple
tubular glands in the section. Large Intestine,
H&E x100.
Fig. 11-22 (below). Simple Branched Alveolar
Gland. Note the branched alveolus or acinus (a)
and the unbranched duct (d). Sebaceous Gland,
H&E x100.

structures called demihmes (of Giannuzzi) the exocrine glands are classified into three
that resemble crescentic caps at the periphery groups: a) merocrtne, 2) holocrine, and, 3)
of the alveoli. The cells in the serous demilunes apocrine.
empty their secretion. into tiny canaliculi that In me-rncrtne glamls, the secretory cells
are located between the mucous cells. The tiny release their secretion by exocytosis. Hence,
canaliculi, in turn, drain into the lumen of the the discharge of the secretion does not result
acinus. in the loss of any part of the cell. Examples of
" merocrine glands are the major salivary glands
and the exocrine portion of the pancreas.
In liolocrine glands, release of secretion
entails destruction of the secretory cells whose
remnants are then discharged by the gland
together with the secretions. The sebaceous'
glands in the skin fall under this category.

Fig. 11-23. Mixed Alveolus (Acinus). Note the

serous demilune that the serous cells form.

Exocrine glands whose secretory units

consist exclusively of serous alveoli are called
serous glands. Those whose secretory units
consist exclusivelyof mucous alveoli are called
mucous glands. Meanwhile, those that contain
serous and mucous, and/or mixed alveoli are
called mixed glands.

Classification of Exocrine Glands According

Fig. 11-24. Mixed Alveolus in a Mixed Gland.
to the Mode of Secretion The serous cells (sc) form serous demilunes at the
According to the mode by which their periphery of alveoli that consist mostly of mucous
secretion is discharged from the secretory cells, cells (mc). Submaxillary Gland, H&E x400.

EPITHELIAL TISSUE _
 In apocrine glands, the apical part of the
secretory cells is released together with the
secretory product. The ceruminous glands
in the skin that lines the external auditory
meatus are examples of this type of gland.
Myoepithelial Cells (Basket Cells)
In the secretory units and small ducts of
many exocrine glands, there are flattened,
stellate cells that are present between the
epithelial cells and the basal lamina. These
cells are called yoepithelial cells. Under
the LM, myoepithelial cells have a flattened,
ESTEBAN & CiONZM.E.S' TEXTBOOK OF HISTOLOGY
--- ----" ---------------------------------------------
dark-staining, fusiform nucleus and scanty
eosinophilic cytoplasm. They possess long
cytoplasmic processes that wrap around a
secretory unit or segment of a duct.
Myoepithelial cells contain actin that is
similar to that found in smooth muscles. These
cells are also contractile. Their contractions
eject the secretions of the acini into the ducts
and propel the secretions towards the main
ducts. --
Myoepithelial cells are present in the sweat
glands, mamm~ry glands, lacrimal glands,
and the major sa~ry glands.
Connective
Tissue

ounecti;v<e{iss-ue, unlike the other

C
basic tissues (i.e., epithelial, muscle and
nervous) which -are highly cellular, is
characterized by an abundance of extracellular
material and a relative paucity of cells.
All the connective tissues in the body are
derived from mesoderm, except for some in the
head that are derived from ectoderm.
The various connective tissues are
classified into two major groups: 'connective
tissue proper and special types of connective
tissue. The latter include cartilage, bone,
blood, and hemopoietic tissue (i.e., myeloid
and lymphoid tissue).
The special types of connective tissue are
discussed in subsequent chapters. The rest
of this chapter, therefore, is devoted to the
discussion of connective tissue proper. Unless
otherwise specified, the term connective tissue
refers only to connective tissue proper.
CONNECTIVE TIS UE PROPER
Connective tissue (proper) is found all over
the body and it can be regarded, as its name
suggests, as primarily the "glue" that binds
body parts together while allowing for some
degree of movement of these parts in relation
to their immediate anatomical neighbors. In
addition, it envelops muscles; forms the stroma
or supporting framework of various organs; acts
as an avenue for the passage of blood vessels and
nerves into and from-the interior of organs and
other parts of the bodyj.serves as a venue for
the exchange of gases and substances between
blood and the other basic tissues; and, provides
the arena and as well as the cells that are needed
to defend the body against invading organisms
and other harmful substances.
Composition of Connective
Tissue Proper
connective tissue, unlike in epithelium, the
cells are not bound to each other. Instead, they
are scattered individually in the extracellular
substance. Furthermore, in connective tissue,
blood vessels and nerve fibers abound in the
extracellular substance.
Extracellular Substance of
Connective Tissue (connective tissue
matrix)
Connective tissue matrix consists of
ground substance and a variety of fibers that
are embedded therein.
Ground Substance
The ground sUDstanceof connective tissue
is an amorphous, homogenous, transparent, and
hydrated gel. It consists mainly of water that
is stabilized by proteoglycans, hyaluronic acid
(hyaluronan), mineral salts, and glycoproteins.
The abundant water in ground substance
makes it easy for oxygen, nutrients and other
needed materials to diffuse from blood to the
connective tissue cells, and for waste products
ofmetabolism to diffuse from the cellsto blood.
Fig. 111-1. Proteoglycan Aggregate. The
illustration shows the organization and
compositionof a proteoglycanaggregate.
the ground substance of connective tissue ~
because of the presence of sulfate and carboxyl
groups in their sugar components.
ESTES/\N & CiONZI\LES TEXTBOOK OF HISTOLOGY
getting attached to hyaluronic acid by "link-
proteins." Hyaluronic acid is a GAGin itself.In
fact, it is the most abundant GAGin the ground
substance of connective tissue. However,
unlike the other GAGs, hyaluronic acid does
not possess a sulfate side-group. Neither does it
covalently attach to a "core-protein." Rather, as
already mentioned, it serves as the backbone to
which proteoglycan molecules are attached by
"link-proteins"to form proteoglycan complexes.
Of the glycoproteins that are present in
ground substance, at least two are fibrillar (i.e.,
they are in the form of microfibrils). However,
they are rather fine such that they are not visible
under the light microscope and are thus, not
classified as connective tissue fibers. The larger
of these two fibrillar glycoproteins is fihrHlin, a
non-sulfated molecule that has a diameter of 10
to 12nm. In electron micrographs,it is seento be
made up of an electron-lucent core surrounded
by an electron-dense area. Fibrillin forms an
integral part of elastic fibers (see page 40). The
smaller of the two fibrillar glycoproteins is
merely 3 to 5 nm in diameter and not much is
known about its structure and function.
The other noteworthy glycoproteins in
the ground substance of connective tissue,
which are probably involved in cell adhesion
and migration, are fibronectin, laminin, and
thFombospondin.
Extrac:ellbJlar Fib rs
The fibers in their extracellular substance
are primarily responsible for the supportive
function of connective tissues. The fibers are
of three types: collagen, elastic, and reticular.
These three types of fibers occur in varying
combinations in the connective tissues that are
present in the different parts of the body.
Collagen Fibers (Collagenous Fibers)

occurring type of connective tissue in the body,
they are the main extracellular fibers.
Individual collagen fibers are colorless in
vivo but when present in abundant amounts, as
in tendons, they impart a white color to fresh
tissue.
Collagen fibers are distinguishable in.
routine histologic preparations because they
are 2 to 10 lAmin diameter. They usually
collect into bundles that appear pink in H&E
preparations because they are acidophilic. To
better demonstrate collagen fibers under the
light microscope, special stains such as Masson's
trichrome that color collagen fibers blue are
sometimes used.
Collagen fibers are made up of collag::en,
.> the most abundant protein in the body and
which accounts for about 25% of the body's dry
weight. The term collagen does not refer to a
single protein; rather, it is the collective term for
a family of structural proteins that have some
commonalities in their molecular organization.
There are currently 28 known distinct types
of collagen, numbered I to XXVIII, that differ
from each other in their amino acid composition
and sequence of polypeptide chains (a-chains). molecules are right-sized by enzymatic removal
Not all the types of collagen are fibrillar in form of the extra amino acids while at the same time,
I~ and not all are present in connective tissue. In three pro collagen molecules (each of which
fact, only collagen types I, II, and III make up is called an a-chain) assemble spontaneously.
Fig. 111-2.Collagen Fibers and Fibroblasts (left).
Photomicrograph shows bundles of collagen fibers
(cf) of various thickness. Scattered among the
collagen fiber bundles are fibroblasts (f).
Fig. 111-3.
Collagen Fiber (below). Electronmicrograph
shows the banding pattern of the fiber: x100,OOO.
practically all the collagen in connective tissue.
Collagen fibers, in particular, are made up of
c_ollagentype I.
Collagen fibers have a tensile strength that
is greater than steel'. They are slightly flexible
but inelastic.
Formation of Collagen Fibers
The molecular precursor of a collagen fiber
is procQl1age_n. It consists of a polypeptide
chain that is longer than those in mature
collagen. In connective tissue, it is synthesized
by fiBroblasts and mesenchymal cells,
which then secrete the said precursor into
the extracellular matrix. (Note: Fibroblasts
and mesenchymal are often said to synthesize
collagen; actually, they are the precursors of
collagen that they synthesize.) Incidentally,
collagen fibers are not exclusively seen in
connective tissue proper. The matrix of
.cartilage and bone also contains collagen fibers.
In these tissues, pro collagen is synthesized by
cells called chondroblasts and osteoblasts,
respectively.
In the extracellular matrix, the pro collagen
CONNECTIVE TISSUE ..
They form trop'ocollage molecules by twisting
around each other, much like the fibers of a
rope, and getting bound together by hydrogen
bonds. Once formed, tropocollagen molecules
aggregate to form tnkrofibrHs.
Microfibrils are fibrillar elements that
are about 45 to 100 nm in diameter each.
Following their formation, they group together
to form bigger fibrillar structures called fibrils
~mac 0 ibdlsY.
Collagen fibrils (macrofibrils) range in
diameter from 0.3 to 0.5 ~m and, as such, are
distinguishable with the use of high quality
light microscope. However, they are better
appreciated in electron micrographs where
they are noted to possess dense transverse
bands that are set at 64 nm intervals along their
length:. Subsequent to their formation, collagen
fibrils group together in p~rallel fashion to form'
collagen fibers:
Elastic Fibers
[Elastic io us are fine fibers that average 1.0
~m in diameter. Unlike collagen fibers, which
usually do not ramify, elastic fibers branch and
anastomose.
When abundant, elastic fibers impart
a yellow color to fresh tissue. In H&E
preparations, they remain unstained and usually
appear as refractile, pinkish-yellow lines that
are difficult to distinguish with certainty from
collagen fibers. When stained with orc in,
however, elastic fibers appear blue to black;
They are also selectively stained by resorcin-
fucnsin and aldeh-x:-de-fuchsindy-e_s..
In electron micrographs, an elastic fiber is
seen as consisting of a homogenous, amorphous
core made up of elastin that is surrounded by
longitudinal bundles of miCrofibril'S) that
consist mostly of ib-rillin
Elastin is a highly insoluble protein that is
responsible for the elasticity of elastic fibers'.
It is resistant to boiling and hydrolysis by
acids, alkali and most enzymes, but it can be
hydrolyzed by the pancreatic enzyme elastase.
ESTEBAN & (iDt'-lZi\LlS TEXTBOOK OF HISTOLOGY
Unlike collagen, which has many genetic
types, elastin has only one. However, elastin in
the body occurs in either of two ways: in the
form of fibers called elastic fibers or in the form
of elastic sheets or lamellae such as those seen
in the walls oflarge and medium-sized arteries.
Elastic fibers are not as widely distributed
as collagen fibers. In mo-st connective tissues,
they are also not as numerous as collagen fibers,
but they ~re particularly abundant in structures
that are subjected to frequent stretching such as
the ligamenta flava between vertebrae. They
are also plentiful in the extracellular spaces of
the elastic cartilages that form the framework
of the auricle and external acoustic meatus of
the ear, external nose, auditory tube, epiglottis,
and some parts of the larynx.
Elastic fibers have lesser tensile strength
than collagen fibers, but in contrast to the latter,
they are very supple. Even when stretched up to
twice their original length, they recoil back to
their original length when the stretching force
is released.
Formation of Elastic Fibers
In connective tissue, the cells that have
the capacity to secrete the substances that are
needed in the formation of elastic fibers are the
ibroblast- and esenchxmal cells. These
Fig. 111-4.Internal Elastic Lamella. The scalloped
black line indicated by the arrow is the internal
elastic lamella, made up of the nonfibrillar form
of elastin. Muscular Artery, Verhoef{ stain x400.
cells, at the start of elastogenesis (i.e., elastic
fiber formation), secrete microfibrils (mostly
fibrillin) into the extracellular space, which,
shortly thereafter, aggregate to form bundles.
Meanwhile, troRoe astin-the precursor
protein of elastin-is likewise secreted by the
same cells into the extracellular space where it
polymerizes to elastin and then incorporated
into the outer aspect of the microfibril bundles.
Subsequently, more and more elastin gets
incorporated into the developing elastic
fiber. When the fiber has enough elastin,
additional microfibril bundles are added on Fig. 111-5.Reticular Fibers (black streaks) and
its external surface. Subsequently, the elastin Reticular Cells (at tip of white arrows) in Reticular
Tissue. Liver, Silver x400.
and microfibrils are re-arranged such that in a
mature elastic fiber, elastin gets to occupy the
core of the fiber while the microfibrils fill the membrane of epithelial and other tissues.
perimeter.
Incidentally, elastin that forms the elastic
Formation of Reticular Fibers
lamellae in arteries is synthesized by ooth Reticular fibers are formed in a similar
muscle cells that lay down elastin in fenestrated manner as collagen fibers. However, the
sheets or lamellae arranged in concentric layers precursors of reticular fibers are synthesized
between layers of smooth muscle. and excreted into the intercellular matrix, not
by ordinary but by specialized fibroblasts called
Reticular Fibers (Reticulin Fibers) reticular cells.
<;Reficular fioeliis are also made up of
collagen. Nonetheless, most authors of histology
Cells in Connective Tissue
and pathology textbooks consider them distinct
from ordinary collagen fibers because their
collagen content is' txpe II while ordinary
collagen fibers, as previously mentioned,
contain collagen type 1.
Reticular fibers are very fine (0.5-:-2.0 ~m
in diameter) fibers that, in contrast to ordinary
collagen fibers, tend to branch and anastomose.
In fact, in some <_?rgans,they form extensive.
networks. They are not distinguishable from
ordinary collagen fibers in H&E preparations,
but they stain black when impregnated with
silver salts. Because of this, they are referred Mesenchymal Cell
to aSIargy:rop'hilic fibers. They also react Mesencnr-ma ce Is are multipotential stem
positively to PAS (periodic acid-Schiff) reagent. cells that have differentiated from pluripotential
Reticular fibers are relatively sparse in cells (see also the chapter on hemopoieis). Like
most connective tissues, but they are the main all multipotential stem cells, mesenchymal cells
extracellular fibers oftrettcular tissue (see page are capable of differentiating into several types
48). They also comprise the fibrillar component of cells. Mesenchymal cells are the stem cells of

CONNECTIVE TISSUE ...

 most connective tissue cells (i.e., fibroblasts accounts for the basophilia of their cytoplasm.
and fibrocytes, reticular cells and adipose They also have irregular cytoplasmic processes.
cells) as well as bone, cartilage and muscle Fibrocytes, on the other hand, are smaller
cells. and have fewer processes than fibroblasts/Their
Mesenchymal cells abound in the embryo cytoplasm is acidophilic and their nucleus is
and in the umbilical cord. They are, however, compact and dark:
rare in adults, but some exist, especially in the There is some amount of functional
bone marrow and in connective tissues near specialization among fibroblasts. Thus,
capillaries. fibroblasts, called reticular cells, that synthesize
There is no general agreement among reticular fibers do not synthesize collagen
experts on how mesenchymal fells lo.o,~like. or elastic fibers. Likewise, fibroblasts that
Most describe them as stellate cells that have synthesize large amounts of glycosaminoglycans
delicate cytoplasmic processes and an oval synthesize little tropocollagen, and vice-versa.
nucleus that contains fine chromatin and a Fibroblasts are capable of mitosis. However,
distinct nucleolus. Under the light microscope, being sturdy and long-lived, they divide
they cannot be distinguished from fibroblasts. infrequently ..
In electron micrographs, they are noted to have
coarser chromatin but fewer and less developed Reticular Cells
cytoplasmic organelles than fibroblasts.
Reticular: cells, as previously mentioned, are
fibroblasts that are specialized to synthesize the
Fibroblasts and Fibrocytes
precursors of type III collagen that make up the
reticular fibers.
in most connective tissues. They synthesize
Reticular cells are slightly larger than
the organic components (i.e., proteins, typical fibroblasts. In H&E preparations, they
glycoproteins, and glycosaminoglycans) of the are noted to have a large and lightly-staining
ground substance of connective tissue matrix nucleus and long cytoplasmic processes that
as well as the precursors of collagen and elastic embrace reticular fibers. They are rather few
fibers. in most connective tissues, but they are the
predominant extracellular fibers in a type of
connective tissue called eticular t' ssue ~see
actively synthesizing connective tissue page 48).
elements). A fibrocyte, under proper conditions
such as in wound-healing, can assume its Adipose Cells (Fat Cells; Adipocytes)
active fibroblast form. In as much as fibrocytes The adiJ20secell is a cell that is specialized
are merely fibroblasts that are in a different to store lipids or fats, mainly triglycerides, in its
functional state, many authors use the terms cytoplasm. It is present in variable numbers in
fibroblast and fibrocyte synonymously. practically all connective tissues. In one type
In routine histologic preparations, of connective tissue called adipuTe tissUE(see
fibroblasts are often seen lying close to or page 47), it is the predominant cellular element.
adhering to collagen fibers. Their ovoid, pale The fat in adipose cells is synthesized by
nucleus contains fine chromatin and a nucleolus. the cells from glucose that is brought to the
Like other active secretory cells, fibroblasts cells from the liver or obtained by the cell from
possess the machinery for the synthesis Of ingested food, via the bloodstream, in the form
proteins including a well-developed rER that ot cnylomicron.

ESTEB/\N& (JCNZl\LES TEXTBOOK OF RISTOLOGY-------

----------
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Fat cells look like fibroblasts before they
accumulate fat or when their fat content has
been depleted. Whe~ a fat cell is only starting
to accumulate fat, at which point it is sometimes
referred to as a lipoblast only a few small fat
droplets in its cytoplasm can be seen. As the
cell accumulates more lipids, the fat droplets
increase in size and number. Subsequently, the'
droplets coalesce to form larger fat droplets,
which further coalesce, until ultimately, a single,
large, fat droplet is formed. The fully-formed
fat droplet in adipose cells pushes and flattens
the nucleus and cytoplasmic organelles to one
side of the cell. Fat droplets are not membrane-
bound; their edges abut on cytoplasm.
Fat cells average SOlAmin diameter. When
they occur singly, they are oval or spherical but
when they occur in clusters, they get compressed
and become polygonal.
In routine LM preparations, the fat droplet
in adipose cells is dissolved and washed away
by solvents (e.g., yJoll during processing.
Consequently, adipose cells in H&E and other
routine preparations simply consist of a large
empty space and a flattened nucleus that is
located on one side of the cell-the reason why
adipose cells are called signet ring cells. Fat is
fixed and stained black by osmium tetroxide and
colored by certain dyes such as SUQanIII.
.' --------, Mature fat cells do not divide. If new or
additional ones are needed, they are sourced
from mesenchymal cells or from Bye-fat cells
' .. p_'« .." •
(greadi_RocIteS). Pre-fat cells are cells that are
..~.;:~ in an intermediate step between stem cells and
.
~~:'~~I fat cells. They reside in adipose tissu~ but their
lineage is uncertain-it is unclear whether they
differentiate from fibroblasts or directly from
mesenchymal cells. They divide twice before
becoming full-fledged fat cells.
Mast Cells (Mastocytes; Histaminocytes)
.,~;ue, note the , ast cells are large, ovoid cells (IS to 20
.' t arrows) and urn) with 'centrally located spherical nuclei
, - ...-;~ fat droplets
and numerous cytoplasmic granules. They
,J.
are practically indistinguishable from other
connective tissue cells in H&E preparations,
but when stained with nluidine blue, their
cytoplasmic granules turn dark purple and
the cells become identifiable under the light
microscope. In electron micrographs, mast cells
are seen to possess microvilli.
The cytoplasmic granules of mast cells
vary in shape and are between 0.3 to 0.8 lAm
in diameter. They are membrane-bound
pouches that contain a variety of chemical
mediators of inflammation notably heparin
(an anticoagulant) and histamine which dilates
and makes blood capillaries more permeable
and which stimulates the smooth muscle cells
especially of the bronchioles in the respiratory
tract and some proteases. '
Mast cells are involved in inflammation
and immediate-type hypersensitivity reaction
(allergic reaction) whose extreme form is
anaphylactic shock. When activated, they
degranulate (i.e., they release the content
of their granules). They likewise synthesize
and release some other substances that also
mediate the inflammatory response but which
are not in their granules such as leukotrienes,
prostaglandins, and certain cytokines.
Aside from their role in allergy and
anaphylaxis, mast cells also playa role in wound
healing and defense against pathogens.
CONNECTIVE TISSUE ..
 Mast cells are derived from cells in the bone
marrow called QJllonr-liorming Unit-Mast
Gell (CEll-. astjJAfter differentiating from its
progenitor cell, a mast cell migrates via blood to
connective tissue where it settles permanently.
In blood, a mast cell probably looks like and
is misidentified as a liasophit, a type of white
blood cell that shares certain similarities with
it In the connective tissues, mast cells live for
weeks to months while retaining their capacity
to divide.'
Mast cells are sparse in most connective
tissues, but they are abundant in the lamina
propria of the gastrointestinal and respiratory
tracts, underneath the skin, and along the course
of small blood vessels.
Macrophages
Macrophages are widely distributed all
over the body. They are present not only in
connective tissues but in practically all tissues.
Macrophages that are in connective tissue
are otherwise known as lUsliocy:tes. Those
in other parts of the body are also referred to
by other names. For examples, those in the
alveoli of lungs are called pulmnnar:y: alveolai
I acropha.ges while those in the sinusoids of the
liver are referred to as Kupffer cells.
Macrophages vary morphologically
depending on their location in the body and
state of activity. Most macrophages are relatively
large cells (usually 25-50 ~m in diameter).
In routine LM preparations, their cytoplasm
is slightly basophilic and typically, appears
ESTEBf\\j 2,GO\ZA..ES'TEXTBOOK OF HISTOLOGY
Fig. 111-7.Pulmonary Alveolar Macrophages.
Macrophages have various names depending on
their location in the body. In the lungs, they are
called pulmonary alveolar macrophages (at tip of
arrows). Lung, H&E x400.
"frothy." Their nucleus is usually indented and
densely staining and possesses a prominent
nucleolus. In connective tissues, macrophages
are morphologically very similar to fibroblasts.
Under the LM, they are easy to recognize only
when they have phagocytosed material in
their cytoplasm. In EM, they are noted to have
numerous lysosomes.
Macrophages ingest and destroy not only
bacteria and exogenous particulate materials,
but also dead or dying cells and senescent tissue
elements. They also playa major role in the
body's non-immune or inflammatory response
(non-immune defense system) by engulfing
and digesting invading microorganisms and
exogenous particulate material. They also help
the body's immune response (immune defense
system) by serving as antigen-presenting cells
(APes) that process and introduce antigens
to lymphocytes (see chapter on the lymphoid
system for further discussion on APCs).
Macrophages, on the basis of their mobility,
are classified as either fixed or free. In connective
tissues, fixe mac.r_ophag~ are attached to
collagen fibers while free macrophages wander
about in the extracellular matrix. A fixed
macrophage is capable of detaching itself and
becoming a free macrophage; conversely, a
wandering macrophage can become a fixed
macrophage. Hence, categorizing macrophages
into fixed or wandering is simply an academic
exercise.
Macrophages are also classified into
either resident or inflammatory. R-esident
macropnage-s are those that inhabit a given site
while Inflammafo ¥ macropnages are thos-e
which, in response to a stimulus, differentiate
from monocytes and migrate to a site.
Resident macrophages are not as
immunologically active as inflammatory
macrophages. They maintain their numbers
in a particular tissue by differentiating from
monocytes or, in as much as they still have
mitotic capability-albeit limited-by local
proliferation. In contrast, inflammatory Fig. 111·9.Plasma Cell (EM). Electron micrograph
macrophages do not proliferate locally; they shows the IIclock-face IIappearance of the
chromatin material (ch) and the highly developed
increase their number by differentiating from
rough endoplasmic reticulum (rER). x5,OOO.
monocytes.
Macrophages can become activated
Most histologists say macrophages have a
and increase their functional activity (i.e.).
life span of about 2 months to 70 days,but more
phagocytic and antigen-processing activity) by
recent evidence indicates that their life span is
certain stimuli. In turn, activated macrophages
much shorter than that, probably 6 to 16 days.
can be deactivated by cert ai n chemical
substances that are produced by the body.
Plasma Cells (Plasmocytes)
A typical plasma cell is slightly bigger than
a red blood cell (RBC), which is about 7.S ~m
in diameter. In routine histologic preparations,
it has a strongly basophilic cytoplasm and an
eccentric nucleus. Its chromatin is present
in clumps that are located near the nuclear
membrane, giving the nucleus a "clock-face"
or "cartwheel" appearance. Its cytoplasm
often exhibits a negative Golgi image. Electron
micrographs of plasma cells often show the
presence of a well-developed and dilated rER,
which attests to the secretory nature of the cell.
PlasmacellsdifferentiatefromWBCs called
Fig. 111-8.Plasma Cell. Note the copious and B Iympliocy:tes (B celis;).They playa major role
highly basophilic cytoplasm, eccentric location of in the body's immune response because they
the nucleus and clumped chromatin in the plasma
produce im-lfiunoglobulins ~antibodiesJ. An
cell (at tip of arrow). Bone Marrow Smear, Giemsa
x1,OOO. antibody: is a substance that is elaborated by

CONNECTIVE TISSUE __

plasma cells in response to the presence of an
antigen (i.e., material that is foreign to the body
like a bacterium, virus, etc.). An antibody binds
to the antigen that triggers its production and
by so doing, it is able to mark the antigen for
destruction by effector cells and/or substances.
A plasma cell is a terminally differentiated
cell that is incapable of cell division or of
reverting back to a B lymphocyte. It has a
lifespan of 10 to 20 days.
Plasma cells are present in limited numbers
in all connective tissues, but they are numerous
.in the connective tissues that are readily
accessible to foreign proteins and bacteria such
as the lamina propria of the digestive tract.
They are discussed further in the chapter on
the lymphoid system.
Leukocytes (White blood cells [WBCs])
teukoc-y:tes or wh-iteb:lood cells refer to the
cells in blood that are not red blood cells. There
.' are several types ofleukocytes: neutro~hilsl
basopnils, eosinopliils, monoc-y.tes and
lym hoeytes. In post-natal life, all leukocytes
are exclusively produced in the bone marrow,
except for the lymphocytes, which are generated
not only in the bone marrow but also in the
various lymphoid tissues and organs.
Mature leukocytes enter the blood
capillaries of the bone marrow and join
circulating blood.' They invariably leave blood
and settle in the connective tissues by escaping'
[STESAf\ (.{CiONZ!\L[S' TEXTBOOK OF HISTOLOGY
........
Fig. 111-10.Dense Irregular Connective
Tissue (region spanned by double arrow).
Note the haphazard arrangement of the
collagen fiber bundles. 'Skin, H&E x100.
from the blood capillaries by amoeboid
movement.
Although present in blood, leukocytes
perform their function in connective tissues.
They are present in variable numbers in
practically all connective tissues. They typically
gather in inflamed areas of the body.
The leukocytes are discussed in detail in the
chapter on blood and hemopoiesis.
Types of Connective Tissue
The connective tissues in the body are
categorized on the basis of their cellular and
extracellular composition into numerous
types such as collagenous, aaipose, r,eticula-l",
elastic, and mucous.
Collagenous Connective Tissue
(Ordinary Connective Tissue)
.Collagenou connective tissue is the most
abundant type of connective tissue in the body .
Itis connective tissue in which the predominant
extracellular fiber is collagen fiber (mllagen
type I) and the predominant cell type is the
fibroblast. Collagenous connective tissue is so
common that it is often referred to as ordinary:
Types of Collagenous Connective.
Tissue
'On the basis of the amount of ground
substance and number and arrangement of the
are, however, innumerable gradations from one
category to the other and within each category.
In many areas in the body, it is sometimes
difficult to classify the collagenous connective __/
tissue that ispresent as either loose or dense. It is,
therefore, not uncommon to encounter phrases
like "relativelyloose" or "relativelydense-when
authors describe collagenousconnective tissues.

Dense Collagenous Connective Tissue Fig. 111-12.Loose Connective Tissue. The pinkish
(Dense Connective Tissue) streaks are collagen fibers while the cells indicated
by the arrows are fibroblasts. Lamina Propria of
Jejunum, H&E x400.

, Dense regular connective tissue, on the

other hand, makes up tendons, ligaments, and
fibrous membranes.

Loose Collagenous Connective Tissue

(Areolar Connective Tissue; Areolar Tissue;
Loose Connective Tissue)

tissue or simply loose connective issue,

is characterized by high vascularity and an
abundance of extracellular substance where the
relatively few collagenous fibers are arranged
haphazardly. It is also more cellular than dense
connective tissue.
Fig. 111-11.Dense Regular Connective Tissue.
Note the regular arrangement of the collagen fiber
bundles and the numerous fibroblasts (at arrows).
Tendon, H&E x 100.

If the collagen fibers in a dense connective

tissue run in various directions, the connective
tissue is called de_n_seirregular, and if the fibers
are arranged ina definite pattern, the connective
tissue is referred to as dense regular.
Adipose Tissue
2\:oipose fissue is a connective tissue in
which the predominant cellular element is the
fat or aaipose cell. Adipose tissue represents
the largest energy storage site of the body. In
normal, well-developedmales, it comprises 12%

CONNECTIVE TISSUE _
to 14% while in females it comprises 20% to 25%
S;?fbodyweight. The fat that is stored in adipose
tissue represents excess dietary calorie intake,
hence, in overnourished individuals, adipose
tissue c~n comprise more than 25% of body
weight. During periods of inadequate caloric
intake, the stored energy in adipose cells is
released in the form ~f fatty acids:
Aside from serving as energy storage,
adipose tissue, particularly that which is stored
in the subcutaneous area; also functions as
thermal insulator. In certain parts of the body,
e.g., soles of the feet and around the kidneys, it
serves as a shock absorber.
The fat cells of yellow adipose tissue store
lipid in the form of a single fat vacuole while the
fat cells of brown adipose tissue store lipid in the
form of numerous droplets.
In the newborn, brown adipose tissue is
present in the neck and back and accounts for
about 2% to 5% of body weight. As one grows
older, however, brown adipose tissue diminishes
in amount such that in adults, it is practically
absent.
Reticular Tissue
Reticular: tissue is a connective tissue in
which the predominant cell and extracellular
fiber are the reticular cell and reticular fiber,
respectively. In reticular tissue, the reticular
cells are usually seen to be attached typically
to the reticular fibers, which form complicated
.networks.
Reticular tissue is abundant and forms the
stroma or supporting framework of the liver,
myeloid tissue, and lymphoid tissues and
organs such as lymph nodes and the spleen.
ESTEBM~& CONZ/\LES' TEXTBOOK OF HISTOLOGY
Fig. 111-13.Mucous Tissue. The tissue is made up
of ajelly-like ground substance where cells, mostly
mesenchymal cells (at arrows), are embedded.
Elastic Tissue
Elastic tissue is a connective tissue in
which the predominant fibrillar component
is the elastic fiber. In this type of tissue, the
.elastic fibers often form bundles that are
arranged parallel to each other. In between the
el~~tic fibers are a few collagenous fibers and
fibroblasts-the predominant cells,
Elastic tissue is typified by the ligamenta
flava of the vertebral .column and the
suspensory ligament of the penis.
Mucous Tissue
Mucous tis_s,ue is characterized by an
abundance of amorphous and jelly-like ground
substance in which a scarce number of collagen,
elastic, and reticular fibers are embedded. The
ground substance of mucous tissue is mainly
composed of hyaluronic acid. Cellular elements
are few and they consist chiefly of mesenchymal
cells and fibroblasts, although an occasional
macrophage may be present .
Mucous tissue is common in the embryo
but rare in adults. It is exemplified by Wharton's
Jelly, the connective tissue present in the
umbilical cord.
 Chapter IV

Muscle Tissue

ontractilitr; the ability to shorten, is an reticulum, sareop-Iasmie rethmlum; and their

C inherent property of all cells. However,

it is exhibited to_ahigh degree by only a
few cells, notablYl~ericytes which are associated
mitochondria, sarcosomes,
There are three types of muscle cells
(fibers): skeletal, smooth, and cardiac.
with very small blood vessels; myoe-pitbelia
Skeletal and cardiac muscle fibers are referred to
cells which embrace the acini and small ducts
as stria ea because, in routine LM preparations,
of some exocrine glands; and usde cells
they exhibit prominent alternating light and
(m:y-oc~tes; m:y.:oidcells) which exhibit t~~ dark cross striations. In contrast, smooth muscle
greatest degree of contractility among all cells cells are nonst .ated because they do not exhibit
in the body and comprise the cellular elements
cross striations.
of muscle tissue.
M'uscle cells are widespread in the body.
M-u-sdetiss-ue,like epithelium and nervous
They occasionally occur Singly or in small
tissue, is a highly cellular tissue. It is composed
disorganized clusters, but in general, the muscle
of muscle cells that are supported and bound
cells in a given area of the body form highly-
together by intercellular material that consists
organized groups that are bound together by
of connective tissue. Muscle tissue is the
connective tissue elements to form muscle
basic tissue responsible for locomotion of the
tissue. There are three types of muscle tissue:
Individual and movement of the various parts
skeletal, smooth, and cardiac, which are made
of the body because muscle cells are highly
up of skeletal, smooth, and cardiac muscle fibers,
contractile. Contraction of muscle cells moves
respectively. These types of muscle tissue differ
or restrains the movement of a body part.
morphologically and functionally.

except for a few organs, notably those in the iri-sl
of the eye that arise from ectoderm. They are
elongated cells-the reason why they are often
referred to as mu-scle fibers instead of muscle
cells-that are individually enveloped by basal
lamina. Their cell membrane is otherwise
known as s4Lcnlemma; their cytoplasm,
sarcoplas-m; their smooth-surfaced endoplasmic
SKELETAL MUSCLE
N early all the skeletal muscle tissue in
the body is organized to form mouse-shaped
organs that are simply referred to as muscles or
skeletal muscles. Most muscles are named, e.g.,
pectoralis major muscle and gastrocnemius
muscle. Incidentally, in medical literature, the

term muscle can refer to either a muscle tissue
or a muscle organ.
Typically, a muscle is attached at either
end by dense regular connective tissue called
tendon to a part of the skeletalsystem (bone or
cartilage), hence the name. The attachments
are referred to as origin and insertion. There
are, however, skeletal muscles whose one or both
ends are not attached to the skeletal system such
as those in the upper third of the esophagus and
the muscles of facial expression.
Skeletal muscle is also sometimes referred
to as :voluntary:muse because it comprises the
muscles of the limbs, body wall, and face that
are responsible for the voluntary movements
of the individual. There are certain areas of the
body, however, such as the pharynx and upper
part of the esophagus, where the skeletal muscle
present is not under voluntary control.
is
Organization of Skeletal Muscle
A typical muscle, the biceps brachii for
example, is a collection of numerous skeletal
muscle fibers bunched in groups called bundles
or fascicles. It is enveloped by tough, dense,
irregular connective tissue called epim,!sium
that keeps the fascicles together.
ESTEBAN & CONZALES' TEXTBOOK OF HISTOLOGY
Fig. IV-1. Organization of a Skeletal
Muscle. A muscle consists of numerous
musclefibers that are individuallywrapped
by delicate connective tissue called
endomysium (en). Within the muscle,
the muscle fibers gather into bundles
or fascicles enveloped by connective
tissue called perimysium (pe) while the
whole muscle is enclosed externally by
connective tissue called epimysium (ep).
Within the muscle, each of the fascicles is
likewise encased by connective tissue called
perimysium. The perimysium keeps the
muscle fibers within the fascicle together. It
also serves as an avenue for the blood vessels
and nerve fibers that supply the muscle fibers in
the fascicle.
Within the fascicle, each of the muscle fibers
is' also individually wrapped and supported,
external to their basal lamina, by a delicate
connective tissue layer called endomysium,
where the _extracellular fibers are mainly
Skeletal Muscle Cells (Skeletal
muscle fibers)
Skeletal muscle cells are long, tapering,
cylindrical, and multinucleated cells that vary
in length «10 to 3S cm) and diameter (10 to 100
urn], They arise in the embryo from the fusion
of mononuclear muscle cell precursors called
my:oblasts that evidently differentiate from
mesenchymal cells.
Skeletal muscle cells have oval nuclei which
vary in number depending on the length of the
cell, but which can total in the hundreds. The
nuclei are longitudinally oriented and located
in the peripheral portion of the cells, near the
sarcolemma. The sarcoplasm of skeletal muscle
 Fig IV-2. Skeletal Muscle, longitudinal section.
Photomicrograph shows a muscle fascicle (f)
enveloped by perimysium (p). The fascicle is made
up of muscle fibers/cells (c) that are encased by
endomysium (e). Note the alternating dark and
light bands and the peripherally-located nuclei.
H&E x200.
cells is acidoFhilic. It contains all the common
organelles-Golgi complex, mitochondria,
endoplasmic reticulum, although rER and
ribosomes are few-and som~ inclusions,
notably lipid droplets and glycogen granules.
In addition, it contains dissolved yoglobin~
an oxygen-binding protein responsible for
the brownish color of the muscle. The most
_noteworthy feature of the sarcoplasm, however,
is the presence of numerous long but thin
filamentous elements called myofi6rils.
Fig. IV-3. Skeletal Muscle, cross section.
Photomicrograph shows muscle fascicles
that are encased by perimysium (p). Each
fascicle consists of muscle cellslfibers that
are enveloped by endomysium (e}.Note
the peripherally-located muscle cell nuclei.
H&E x200.
MyofibrUs
There are roughly 5,000 to 10,000
myofibrils per muscle fiber, each of which spans
the length of the cell. Each of the muscle cells
that comprise a muscle also spans the length
of the muscle. The myofibrils vary in diameter
from 1 to 2~m. They are arranged parallel to
the long axis of the cell and exhibit transverse
striations of alternating light and dark: band_so
The light and dark bands of one myofibril are
aligned with the corresponding light and dark
bands of the other myofibrils in the same muscle
cell. Hence, when viewed in longitudinal section
under the light microscope, a skeletal muscle
cell looks striated. The light bands are called
isotropic bands (I-Banas) because they do
not alter polarized light. In contrast, the dark
bands are called anisotropic banits (A-bands)
because they display birefringence in polarized
light.
Fig. IV-4. Skeletal Muscle (EM). The electron
micrograph shows the different regions of a
myofibril including the I-band (I), A-band (A), z-line
(Z), H-band (H), and sarcomere (S).
MUSCLE TISSUE

- .
_.-.---
l The filaments
Fig. IV-S.Composition and Organization of a
Myofibril
In electron micrographs (and occasionally,
in well-prepared light microscopic specimens)
several other distinct zones can be appreciated
further in a myofibril. A fine dark transverse line,
. the l;-line ~Zwisdj.enseh:eillenline, -lland, or
-(lise), bisects the f-band. The A-band, on the
other hand, has a lighter mid-portion called
H-ban{f ~Heller band~ that is further bisected
by a thin dark stripe, the -line (Mittels~fiei1>e
line) "
Sarcomere
A myofibril is made up of numerous
(up to 10,000) small contractile units called
sarcomeres that are laid end to end. A sarcomere
refers to the region that spans two Z-lines and is
about 1.5 to 2 ~m long in a resting muscle.
ESTEBil.N& CiONZ/\LES'TEXTBOOK OF HISTOLOGY
A sarcomere, in turn, has been shown by
high resolution electron microscopy to consist
of a collection of thread-like structures called
filaments (my:ofilame-nts). There are around
1,000 to 2,000 of them that are arranged parallel
to the long axis of the sarcomere.
Muscle Filaments (Myofilarnents)
(myofilaments) that
comprise a sarcomere are of two types: thick
and thin.
The thick frlarrrerrts occupy the middle
zone of a sarcomere. They span the region of the
A-band. They are 10 to IS nm in diameter and
1.5 to 1.6 ~m in length. They are kept aligned by
the attachment of their midpoints at the M-line.
The thin filaments, on the other hand,
occupy the peripheral zones of a sarcomere.
They are more numerous, but are only about
half as thick (5 to 6 nm) and are shorter (length
1.0 urn) than the thick filaments. One end of
each thin filament is attached to a Z-line while
the other end is free (see Fig.N-6).
In the resting muscle cell. the thick and
thin filaments partially overlap each other
at the A-band. A cross section of this overlap
reveals that each thick filament is surrounded, in
hexagonal pattern, _bysix thin filaments. In the.
central region of the A-band, the thick filaments
are not overlapped by the thin filaments.
This region corresponds to the H-band. The
peripheral areas of adjacent sarcomeres, in
contrast, are occupied only by thin filaments.
These areas comprise the I-band.
Fig. IV-6.The Arrangement
of the Myofilaments in a
Sarcomere

Fig. IV-7. Movement of the Thick and Thin
Filaments with Respect to Each Other during
Muscle Contraction. When a muscle contracts,
the thick filaments in a sarcomere remain stationary
while the thin filaments move towards the center
of the sarcomere.
Proteins in Muscle Filaments
The filaments of skeletal muscle cells
are made up mainly of four proteins: actin,
tropomyosin, troponin, and myosin. Of these,
actin and myosin are the most abundant,
accounting for about 60% of total muscle
protein. The thin filaments consist of actin1
tro~omxosin, and troBonin while the thick
filaments consist of }':os'n.
The principal protein component of the
thin filaments is F-actin As mentioned in
Chapter I, it is the filament form of actin that
consists of two "strands of globular and soluble
actin f6-acti-nJ molecules coiled around each
other, much like the fibers of a rope, to form a
filament. To keep the actin filaments aligned,
F-actins are anchored to the Z-line by proteins,
notably a-actinin and desmin. Each of the two
G-actin molecules that makes up a thin filament
possesses binding sites for myosin.
The other two protein components of the
thin filaments, tropomyosin and troponin, form
what are called the troponin-tropom-y-osin
complexesthat are arranged along both sides of
each actin filament. The troponin-tropomyosin
complexes playa role in the regulation of
contraction.
The thick filaments, on the other hand,
consist of myosin molecules. Myosin makes up
60% (in contrast, actin comprises only 15%) of
the total proteins in myofibrils.
A myosin molecule is much bigger and
heavier than an actin molecule. It is composed
of six polypeptide chains, two heavy chains,
and four Ii ht chains, which are so arranged
as to form a structure that has two heads and
a tail. One end of each of the two heavy chains
twists with the other to form the tail while each
of the two heads is formed by the other end of
a heavy chain together with two light chains.
Each head has a binding site for actin.
A thick filament consists of about 274
myosin molecules that are assembled in such a
way that their heads project at regular intervals
along the filament.
Mechanism of Skeletal Muscle
Contraction
The widely accepted explanation for the
mechanism of muscle contraction is the sliding
filament tneoq:. This theory states that when
a muscle contracts, it shortens because the
interaction of the actin and myosin molecules
causes the thin and thick filaments to slide past
each other, resulting in the shortening of the
sarcomeres.
During muscle contraction, within each
sarcomere, the thick filaments remain stationary
while the thin filaments get pulled by the thick
filaments towards the center of the sarcomere.
Consequently, the overlap between the thin and
thick filaments increases, causing a progressive
diminution in width of the 1-and H-bands. The
Z-lines, meanwhile, move towards the center
of the sarcomere because they get dragged by
the thin filaments attached to them. Since all of
the sarcomeres shorten while the origin of the
muscle remains stationary, the insertion moves
towards the origin.
MUSCLE TISSUE ..
 How do the thick filaments pull the thin
filaments? As mentioned earlier in this chapter,
the heads of the myosin molecules contain
binding sites for actin molecules while the
actin molecules have corresponding binding
sites forthe heads of the myosin molecules. In
the resting muscle, these binding sites cannot
interact because those in the actin molecules are
covered by troponin-tropomyosin complexes.
The binding sites in the actin molecules
can, however, be uncovered by calcium ions
because calcium ions bind with the troponin-
tropomyosin complexes, causing the complexes
to shift position, thus exposing the binding sites.
Hence, to initiate muscle contraction, calcium
ions have to be released into the area where the
troponin-tropomyosin complexes are.
As soon as the binding sites in an actin
molecule are exposed by calcium ions, the
heads of the myosin molecules promptly and
spontaneously bind with them. This binding
triggers the hydrolysis of ATPs by the ATPases
that are present in the myosin heads. It results
in the release of energy, an event that could
not happen earlier because myosin needs actin
as a co-factor in order to hydrolyze ATP. The
energy that has been generated enables the
heads of the myosin molecules to bend or flex,
pulling the thin filament (i.e., actin molecule)
towards the center of the sarcomere. The
myosin heads remain in flexed position until
new ATP molecules bind to them and get
hydrolyzed. Thereafter, with the energy that
has again become available, the myosin heads
are able to disengage, recoil back to their former
positions, reattach themselves to other binding
sites in the actin molecules, and repeat the
bending action they performed earlier. These
movements of the myosin heads are repeated in
a rapid fashion until the thick (myosin) and thin
(actin) filaments have completely overlapped.
Transverse Tubules (T-tubules)
and Sarcoplasmic Reticulum
The sarcolemma of striated muscle cells
forms tubular invaginations that penetrate the
EST[8;.\N & GONZ/\LES' TEXTBOOK OF HIST0LOGY
muscle fiber and create anastomosing systems
of tubes that surround the sarcomeres of the
myofibrils at the junction of the A- and l-bands.
These tubes, whose lumens are continuous with
the extracellular space, are called Itransverse
tubules (T-tubulesY.
Meanwhile, in the area of the myofibrils
in between the T-tubules, the sarcoglasmic
reticulum (i.e., sER:1creates an intricate and
complex system of membrane-bound channels
whose function is to capture and store calcium
ions needed for muscle contraction. At the
junction of the A- and l-bands, the membrane-
bound channels form a pair of large, flattened
cisternae that are closely applied to either side of
a T-tubule. These cisternae are called terminal
A T-tubule and the pair of terminal cisternae
associated with it are collectively referred to as
a triaa.
When the sarcoplasmic reticulum
depolarizes, it releases its store of calcium ions
into the area of the overlapping A- and l-bands.
This initiates a series of events-described in
connection with the sliding filament theory-
that culminates in muscle contraction. When
depolarization ends, the sarcoplasmic reticulum
acts as a drain that enables calcium ions to
return into the cisternae.
The signal for the sarcoplasmic reticulum
to depolarize comes from the motor endpla
on the surface of the muscle cell. Although the
myofibrils are not equidistant from the cell
surface, they are able to contract simultaneously
because the depolarization impulse is
instantaneously transmitted to the terminal
Cisternae via the T-tubules.
Motor Endplate (Myoneural
junction; Neuromuscular junction)
The command for a skeletal muscle to
contract is a neural one. It originates from the
central nervous system and is carried to the
surface of the muscle cells by the myelinated
nerve ffbers (i.e., axonsy of somatic motor
 Fig. IV-S. Motor Endplate
somatic motor neuron arborizes and forms
numerous bulb-like terminations (axon
terminals). These terminals make contact with
the sarcolemma of up to 160 muscle fibers. A
somatic motor neuron, together with the muscle
fibers it supplies, comprises a mntor unit.
At their point of contact, an axon terminal.
and the sarcolemma of a skeletal muscle fiber
form a specialized structure called motor
q:ndpla e. Each muscle fiber has only one motor
endplate.
In the motor endplate, an axon terminal
sheds its myelin and settles into a depression on
the surface of the muscle fiber called s}'napthc
Itrougll or ~-rimary synaptic cleft. In the
primary synaptic cleft, the axolemma and the
sarcolemma are separated by a narrow space
that is merely SO nm wide. The sarcolemma
in the primary synaptic cleft further forms
numerous deep folds called junctional folds
(seco dary s~na12ticcle(ts~perpendicular to
the primary synaptic cleft. In these folds, the
muscle cell exhibits an accumulation of nuclei
and numerous mitochondria, ribosomes,
and glycogen granules in its sarcoplasm. The
expanded axon terminal, meanwhile, also has
numerous mitochondria and avery large number
of vesicles that contain the neurotransmitter
acetylcholine.
Upon the arrival of the nerve impulse for
muscle contraction at the axon terminal, the
neurotransmitter-containing vesicles release
their acetylcholine content into the synaptic
cleft. Acetylcholine then diffuses across the
synaptic cleft and triggers local depolarization
that rapidly spreads across the surface of the
muscle fiber, the T-tubules, and sarcoplasmic
reticulum, which then releases calcium ions to
start muscle contraction.
The action of acetylcholine terminates
when it is degraded by the enzyme
acetylcholinesterase.
Types of Skeletal Muscle Fibers
There are three types of skeletal
muscle fibers that vary morphologically
and functionally. These are red, white, and
intermediate.
Red masEle fibers are smaller and have
a richer blood supply than white muscle
fibers. Furthermore, their sarcoplasm has
more mitochondria, glycogen granules, and
myoglobill'
muscle fihers because they contract at a slower
rate than white fibers. White muscle fibers, on
the other hand, are called fast twitch muscle
fibers ecause they contract at a faster rate than
red muscle fibers. Their contraction is also more
MUSCLE TISSUE ..
forceful than that of red muscle fibers.but
fatigue faster.
muscle fibers.
Most human skeletal muscles contain a;
mixture of the three types of muscle fibers, but
the ratio of one fiber type to another depends on
the function of the muscle. For example, the long
muscles of the back and the postural muscles of
the neck and leg are made up predominantly of
red muscles, while the muscles used for running
such as the gastrocnemius are composed mostly
of white fibers.
A typical skeletal muscle consists of
numerous motor units. Although a skeletal
muscle contains a mixture of all three types
of skeletal muscle fibers, .the .muscle fibers that
comprise a motor unit are of the same type.
Furthermore, the motor units that comprise
a skeletal muscle are often not activated
-sirnultaneously, For example, if only weak
contraction is needed, only the motor units
consisting red muscle fibers are activated.
There is also evidence which shows that
exercise can transform the skeletal muscle fibers
from one type (e.g., intermediate) to another
(e.g., white).
Proprioceptive Organs in Skeletal
Muscles
Skeletal muscles, their tendons, and the
ESlEB/-\f-.J& CCNZ!\LES' TEXTBOOK OF HISTOLOGY
they are receptors whose collective function is
proprioception, i.e., the monitoring of the
position of the limbs and state of contraction of
the muscles.
Neuromuscular Spindle (Muscle
Spindle)
Neuromuscular spindles embedded in
the endomysium and perimysium-are present
in all skeletal muscles. However, they are
particularly numerous in muscles involved in
fine motor movements such as the extra-ocular!
m scles.
A neuromuscular spindle is an encapsulated
fusiform structure that is between 1 to 6
mm in length. Its capsule which consists of
connective tissue encloses a fluid-filled space
that contains several modified striated muscle
fibers (int fusa fibe s) which are smaller and
shorter than the surrounding skeletal muscle
fibers (extrafusal fibers). There are two kinds
of intrafusal fibers: nuclear bag and nuclear
chain. NUJ:_Lea_ bag --'-bes possess a dilated
central area that contains a bunch of nuclei. The
I •
uclear chain fiber- , on the other hand, do not
manifest any dilatation. Moreover, their nuclei
are set in a single row.
The intrafusal fibers are provided with
two types of se~-S-_{)J:~ nerve endrngs. One
type, the annulos~~ral ending, consists of the
unmyelinated terminations of sensory neurons
that are spirally wrapped around the central
portion of the intrafusal fibers. The other type,
the flower-sBray; ending, consists of smaller
nerve endings that innervate the peripheral
portions of the intrafusal fibers.
The neuromuscular spindle is a stretch
receptor that detects the degree and velocity of
stretch applied to a muscle.
Golgi Tendon Organ
The 60lgi tendon organs are small, one-
mm-long structures located in the tendons that
attach skeletal muscles to their insertions and
origins. They consist of c_oJlagenfibers enclosed
Fig. IV-9. Cardiac Muscle, cross section .. Note 'that the muscle cells=which are individually wrapped
by endomysium (e)-form fascicles that are enclosed by perimysium (p). Also note the central location
of the nuclei (at tip of arrows) of the muscle cells. Heart, H&E x400.

by a thin, cone-shaped connective tissue striated and its contractions are forceful. Unlike
capsule. Each of them is supplied by a single most skeletal muscles, however, it is not under
afferent nerve fiber that discards its myelin and conscious control.
breaks into branches as it enters the capsule.
Within the organ, the slender nerve endings are Organization of Cardiac Muscle
in between the collagen fibers.
Cardiac muscle tissue consists of cardiac
The Golgi tendon organs are sensitive to, , muscle fihers that form bundles or fascicles.
muscle contraction rather than stretch. They The muscle fibers and fascicles in ~ardiac muscle
evidently measure the tension that is generated are organized much like those in skeletal muscle.
by muscle contraction. Each fascicle is surrounded by Berill1}':si:urn
The muscle fibers, in turn, are individually
C PIAC MUSCLE enveloped, external to their basal lamina, by a
thin ~d_om:y:sium.

heart and sometimes, in small areas in the wall
of some of the big blood vessels attached to the
heart. Like skeletal muscle,
, cardiac muscle is
Fig. IV-10. Drawing of Cardiac Muscle Cells
Showing Intercalated Discs (at arrows)
Cardiac Muscle Cells (Cardiac
muscle fibers)
Cardiac muscle cells are cylindrical cells
that are much shorter than skeletal muscle cells.
Their average length is only SOto 100 l1m, but
with their average diameter of IS l1m, they are
as broad as many skeletal muscle cells. Unlike
skeletal muscle cells which do not branch,
cardiac muscle cells typically split longitudinally
at their ends to give off a few branches.
Whereas skeletal muscle cells are
multinucleated, cardiac muscle cells contain
only one to two nuclei. Furthermore, in contrast
to skeletal muscle cells whose nuclei are in the

MUSCLE TISSUE ..
Fig. IV-11. Cardiac Muscle,
longitudinal section. Note the
centrally-located nuclei (nu) and
the presence of intercalated discs
(at arrows). Heart, H&E x400.

periphery of the cell, the pale-staining nuclei in cardiac muscle cells surround the Z-lines.
of cardiac muscle cells are centrally located. Also, in cardiac muscle cells, the lumens of the
Also, the sarcoplasm of a cardiac muscle cell is T-tubules are bigger.
more abundant than that in a skeletal muscle The sarcoplasmic reticulum of cardiac
cell. Its mitochondria are also more numerous muscle cells is not as well developed as that of
and larger. But, as in skeletal muscle cells, the skeletal muscle cells. In many instances, only
sarcoplasm of a cardiac muscle cell is filled with one expanded terminal cisterna is associated
myofibrils. with a Ttubule, thus, d")'"ads,instead of triads,
are formed.
Myofibrils of Cardiac Muscle Fibers
The myofibrils in cardiac muscle fibers, Intercalated Discs
like those in skeletal muscle fibers/consist Cardiac muscle cells have distinct cell
of sarcomeres that are laid end to end. The boundaries. However, they appear to form
sarcomeres likewise contain thick and thin
syncytium under the light microscope because
filaments (myofilaments) which have the their terminal branches are at ached end-to-
same protein composition (with a few minor end to the terminal branches of their neighbors
differences)and arrangement asthose in skeletal
via specialized junctional complexes called
muscles. However, owing to the abundant
dntercalated discs.
sarcoplasm and numerous mitochondria
in cardiac muscle cells, the cross striations Intercalated discs are unique to cardiac
of their myofibrils are not as prominent as muscle.Under the light microscope, they appear
those of skeletal muscle cells under the light as dark, transverse lines that occur at irregular
microscope. intervals.
In electron micrographs, an intercalated
T-tubu,'es and Sarcop,'asmic Reticulum disc exhibits two regions: atransverse portion
of Cardiac Muscle Fibers and a lateral portion.
The sarcolemmaofcardiac muscle cells,like The transverse portioN contains two forms
their skeletal muscle counterparts, invaginates of junctional complexes, fasci adherens and
to form T-tubules. But unlike in skeletal muscle desmosome. A fascia adherens is similar to the
cells where the T-tubules are disposed at the zonula adherens of epithelial cells, except that
junctions of the A- and I-bands, the T-tubules the former does not form a band that encircles

ESTEBAN & (iCP~Z/\LES' TEXTBOOK OF HISTOLOGY

the cell but is instead a broad intercellular
junction. The thin filaments of the terminal
sarcomeres of a cardiac muscle cell attach into
the fasciae adherens. Thus, in a cardiac muscle
cell, the fasciae adherens in the intercalated
discs can be regarded as the terminal Z-lines.
The desmosomes, on the other hand, which are
present at regular intervals, are identical to the
desmosomes that bind epithelial cells.
The lateral I2ortion of the intercalated discs,
meanwhile, runs parallel to the myofilaments.
It is characterized by ga~ junctions) that are
likewise identical to those between epithelial·
cells,
Thus, the transverse portion of the
intercalated discs serves to anchor the
myofibrils and to keep the cells together.
The lateral portion, in contrast, allows for
instantaneous spread of contractile stimuli from
one cell to another.
Mechanism of Cardiac Muscle
Contraction
The chain of events that occurs during the
contraction of cardiac muscle cells is similar to
that of skeletalmuscle cells.However,in cardiac
muscle cells, the calcium ions that uncover the
binding sitesin the actin molecules to enable the
binding sitesin the myosin molecules to interact
with them come not only from the sarcoplasmic
reticulum but also from outside the cell. Also,
unlike skeletal muscle cells, cardiac muscle
cells contract without neural stimulation.
The impu se that initiates their contraction is
generated by the 'sinoatrial node or S:A:node,
a small structure in the heart that consists of
urkinj-efibe s.
Purki nje fibers are modified cardiac
muscle cells. They are non-contractile cells
that are specialized, to comprise the impulse
conducting system of the hearrwhich generates
and propagates the electrical impulse that
initiates cardiac contraction. The impulse
conducting system of the heart, of which the
SAnode is a part, is discussed in the chapter on
the circulatory system.
Although they do not need a neural stimulus
to contract, cardiac muscle cells are nevertheless
supplied with efferent fibers by the motor
neurons of the autonomic nervous system.
These efferent fibers serye to regulate the rate
and strength of cardiac muscle contraction. The
axon terminals of these efferent fibers end a
short distance from the muscle cellsthey supply.
On arrival of an efferent stimulus, the axon
terminals release their neurotransmitters into
the intercellular space. The neurotransmitters
reach the receptors on the surface ofthe muscle
cells by diffusion.
SMOOTH MUSCLE
Smooth muscle (tissue)iswidelydistributed
in the body. It comprises the muscular
component of the w I o_yis_c_eral organs-
which is why it is otherwise known as visceral
muse e-and hlooil=v:essels.It is also present
in the pare-nchy_maof most internal organs
and even the skin. Smooth muscle is sometimes
referred to as involuntary muscle -because it is
not under conscious control. However,the term
is inappropriate because cardiac muscle and
skeletal muscle in certain areas of the body are
also not under conscious control.
The contractions ofsmooth muscle are slow
and not as forceful as those of striated muscles.
Fig. IV-12. Smooth Muscle, longitudinal section.
Note the fusiform nuclei that are centrally-located
within the cells. Colon, H&E x400.
MUSCLE TISSUE 'U
Organization of Smooth Muscle
Smooth muscle cells sometimes occur
singly or in disorganized clusters, but more
often, those in an area or organ are organized
to for~ bundles (fascicles) enveloped by
perimysium. The number of cells that comprise
a fascicle varies from a few to hundreds. In
the fascicles, as in striated muscle tissue, the
smooth muscle cells are individually enveloped,
external to their basal lamina, by endomysium.
Furthermore, the cell membranes of adjacent
cells in smooth muscle fascicles are attached to
each other by desmosomes and gap junctions
that are similar to those seen between epithelial
cells.
The arrangement of- smooth muscle
fascicles varies depending on their location.
In the walls of the gastro-intestinal tract, for
example, they form two layers: one layer consists
of longitudinally-oriented fascicles; while
the other layer consists of circumferentially-
arra~ged muscle fascicles. In contrast, the
fascicles in the urinary bladder form three ill-
defined layers, two layers of longitudinally-
arranged muscle fibers that sandwich a layer of
circularly-arranged muscle fibers.
Smooth Muscle Cells (Smooth
muscle fibers)
Smooth muscle cells are fusiform cells that
are broad in the middle and tapering at both
ends. They vary greatly in length dependi~g on
the organ where they are located. They may be
as short as 20 ttm such as those that are in the
walls of the small blood vessels, or as long as
500 ttm such as those in the pregnant uterus.
Their diameter is generally between 2 to 10
ttm. They contain a single, oval nucleus located
in the thick part of the cell. Their sarcoplasm
is acidophilic. It is filled with filaments
(myofilaments) that consign the cytoplasmic
ulganelies (mitochondria, ribosomes, rER,
Golgi complex) to the perinuclear area.
Smooth muscle cells, when seen in
longitudinal sections under LM, may be
ESTEB/'>_f\j & GO\Z/\!...[STEXTIBOOKOF HISTOLOGY
mistaken for collagen fibers because, despite the
presence of numerous filaments (myofilaments),
their sarcoplasm appears homogenous. Collagen
fibers, however, are less acidophilic and have
no nuclei. The nuclei seen among the collagen
fibers in histologic sections belong to fibroblasts.
In the smooth muscle fascicles, the smooth
muscle cells are arranged parallel to each other
with the thick part of one cell lying on the thin
parts of neighboring cells. Hence, when seen
in cross section under the light microscope,
adjacent smooth muscle cells exhibit different
diameters and only the larger ones that are cut at
the level of the thick portions show the presence
of a nucleus.
Filaments (Myofilaments)
As in striated muscle cells, the filaments
(myofilaments) in the sarcoplasm of smooth
muscle cells are of two types: thick and thin.
However, the ratio of thin to thick filaments is
much higher in smooth muscle (15:1) than in
skeletal muscle (6:1).
Similar to striated muscle, the thick
filaments of smooth muscle cells consist of
myosin while the thin filaments are made up
mostly of actin. In smooth muscle, the thick
filaments contain much less myosin and the thin
filaments do not contain troponin.
Unlike striated muscles, the filaments
(myofilaments) of smooth muscle cells do not
form sarcomeres, much less myofibrils-that is
why the cells are not striated. Instead, the thick
filaments are scattered all over the sarcoplasm
while the thin filaments which surround
the thick filaments are anchored on dense
bodies that contain the protein a-actinin and
correspond to the Z-disks of the myofibrils of
skeletal muscle cells. Some of the dense bodies
are attached to the sarcolemma but most are
in the cytoplasm anchored on a network of
intermediate filaments made up of the protein
desmin.
Smooth muscle cells have a poorly
developed sarcoplasmic reticulum that forms
 Fig. IV-13. Smooth Muscle,
cross section. Note that in
the 'cells where the nucleus
is seen, the nucleus is
centrally located. The arrow
indicates endomysium that
envelops each muscle cell.
Duodenum, H & E x400.

a loose network throughout the sarcoplasm. ATP breakdown releases energy that enables
Furthermore, their sarcolemma does not form the myosin molecule to interact with actin
T-tubules. molecules.
Smooth muscle cells do not need neural
Mechanism of Contraction of stimulation to contract. They are inherently
Smooth Muscle contractile, although in some visceral organs
such as the small and large intestines, there are
In smooth musclecells,asin striated muscl~.
pacesetter cells (interstitial cell ofCajal; ICC)
cells,contraction results from the interaction of
that trigger the contraction of the muscle cells.
myosin with actin molecules. Thus, in smooth
muscle cells,thin filaments also slidepast thick Like cardiac muscle cells, smooth muscle
filaments. However, since the dense bodies cells are supplied with efferent fibers by the
where the thin filaments are attached do not autonomic nervous system whose axon
form a straight line, shortening occurs in all terminals end a short distance from the cells
directions. they supply. The efferent fibers release their
neurotransmitters into the intercellular space.
Like in striated muscles, calcium ions also The neurotransmitters reach the receptors on .
regulate smooth muscle contraction, although the surface of some of the smooth muscle cells
in a slightly different manner. Most of the by diffusion and then propagated to the other
calcium ions that trigger contraction of smooth cellsvia gapjunctions.
muscle cellsdo not come from the sarcoplasmic
reticulum, but from the extracellular substance.
DREGE E~ 0
They enter the cell when the cell surface
depolarizes. Once inside the cell, calcium
USCLE TISSUE
ions interact, not with troponin-tropomyosin Skeletal muscle tissue is capable of some
complexes because there are none, but with degree of regeneration despite the fact that
an enzyme complex on the myosin molecule, skeletal muscle cells are incapable of cell
calmodulin-myosin light chain kinase. This division. The source of new muscle cells is
interaction activates the enzyme myosin a small residual population of myoblast-like
light chain kinase which breaks down ATP. stem cells called satellite cells within the

MUSCLE TISSUE __
 /
/
basal lamina that surrounds the muscle cells.
These cells, which decrease in number with
age, cannot be distinguished under the light
microscope. Following injury to a muscle, they
probably divide then fuse together to form new
skeletal muscle cells. The regenerative capacity
of skeletal muscle, however, is very limited.
Often, if the injury to the tissue is massive,
the dead muscle cells are replaced not by new
muscle cells but by connective tissue elements
that form a scar.
Skeletalmuscle cellsprogressivelydecrease
in size and number starting at age 25.
[STEB/'I}~& CiO\jZA_ES' TEXTBOOK OF HISTOLOGY
The regenerative capacity of smooth
muscles, on the other hand, depends on
their location in the body. In some organs,
smooth muscle cells have minimal to absent
regenerative capacity such that when they are
lost, they are replaced by connective tissue
elements.In other organs, ofwhich the pregnant
uterus is the extreme example, numerous new
muscle cells can be produced when needed.
In contrast to smooth and skeletal muscles,
cardiac inuscle has negligible regenerative
capacity. Cardiac muscle cells that die are
invariably replaced by connective tissue
elements.
 Chapter V
Nervous Tissue
N
erxous tissue, like epithelial tissue,
is made up of erosely packed cells that
are separated by very little amount of
intercellular substance. It is, in fact, considered
by some authors as simply a special type of
epithelial tissue because of its high cellularity
and because it arises from embryonic
e€toderm.
The nervous tissue in the body is organized
to comprise the neJ;¥OUSsystem, which is
anatomically divided into two divisions:
central nervous system '€NS~which refers to
the brain and the spinal cord, and peripheral
nervous system ~NS) which relates to all
other nervous tissue in the body.
Nervous tissue in the eNS is devoid of
connective tissue except for those associated
with blood vessels. It simply consists of a mass
of cells that abut on each other. In the PNS
however, there is some amount of intercellular
material, mainly connective tissue, albeit
minimal.
CELLS OF NERVOUS TISSUE
The cells of nervous tissue consist of
neu ons (ne e cells) and supporting cells
(neuroglial or glial cells).
Neurons (Nerve Cells)
Neurons are the functional units of nervous
tissue-they perform all the functions of the
tissue. They are highly specialized cells that
exhibit irritability (i.e., the ability to react to
stimulus) and conductivity (i.e., the ability to
transmit stimulus) to a high degree. Within the
body, neurons are arranged in close apposition
to, and communicate extensively with, each
other.
There is no consensus among experts as
to the total number of neurons that comprise
the nervous system. Estimates vary between 14
billion and 1trillion.
Neurons are the most morphologically
variable cell type in the body. In terms of size,
most neurons are large cells (up to 150 um in
diameter) and some are, in fact, large enough
to be seen by the naked eye. However, there are
many small neurons such as the granule cell in
the cerebellum which is only 5 l1min diameter.
Neurons come in all sorts of shapes. The
following are some examples: steHate:neurons
characterize the ventral gray matter of the
spinal cord and the motor nuclei of the brain
stern: pyrarrridal eurons are present in the
cerebral cortex: and flask-shaped neurons called
Purkinje cells that give off a dendrite which
arborizes like a tree are seen in the middle layer
of the cerebellar cortex. Many more neurons
.that vary in shape are found in the cerebellar and
cerebral cortices.

Parts of a Neuron
Although highly variable in form, neurons
have things in common. They all consist of a
cell body called perikaryon or so~~ a"iidone
.or l1}()reprocesses that-draw out from the nerve
cell body.'
The processes of neurons are of two kinds:
axon and dendrite. The number and location of
Fig. V-1. Parts of a Myelinated Neuron
these processes, especially the dendrites, largely
determine the shape of the neuron. ..
Cytoplasmic Organelles
Neurons are terminally differentiated cells
Neurons contain the same cytoplasmic
that are incapable of cell division'. In adults,
organelles present in most other cell types
very few neuronal stem cells persist. That is the
including an endoFlas~ic reticuhufi,
reason why in general, when a neuron dies, it'
G-olgi compJex, ibosomes, mitochonilFia,
is n~t replace"G.However, axons~ and dendrites
,. f;".~ __
li)1_:sosomes,
peroxisoni'es, and a centrosome.
can regenerate -tv ien Hamaged,providetl tIle cell
body is intact. ., - - In neurons, the rER is highly developed. In
routine LM preparations, parts of the neuron's
Perikaryon rER are seen as deeply basophilic, granJ!lar
masses that are referred to as Nissl bodies
The cell body or perikaryon of a neuron
Echromophilic substances; tigroid bodiesJ"
consists of a nucleus that is surrounded by
Nissl bodies are aburrdant throughout the
basophilic cytoplasm G europlasm~ and
enclosed by a cell membrane (neurolemtna~ perikaryon and are also found in dendrites, but
that likewise envelops the processes of the cell. they are notably absent in the axon and the axon
Neurolemma is structurally the same as the cell hillock, i.e., the area of the perikaryon where
membrane of other cell type'S. the axon originates. The number, form, size, and
distribution of the Nissl bodies vary depending'
Like other cell types, a neuron has
on the neuron. ha.y.al:e:usuallylarger and more
cytoplasmic organelles, a cytoskeleton, and
numerous in the bigger neurons.
inclusions.
A Golgi complex is present in all neurons,
Nucleus but is confined to the £erika-ryo~.
Usually, neurons have only one nucleus, In neurons, as in other cell types, the rERs
but some have more. The nncleus of a neur.QOlis (Nissl bodies) and Golgi complex are involved
typically large, spherical or ovoid, and centrally in the synthesis of proteins essential for the
located. Its nuclear enveleEe is structurally maintenance of the structural and metabolic
similar to that of other cells while its chromatin integrity of the cell. In addition, they synthesize
material is finely dispersed. Hence, in routine the protein component of the neurotransmitters
LM preparations, the nucleus is usually pale and that are used by neurons in communicating with
the nucleolus very prominent. other neurons and cells.

ESTEB!\f'~& CONz/\LES' TEXTBOOK OF HISTOLOGY

 Mitochondria are abundant in neurons, Inclusions
but they are generally smaller than those seen
A variety of inclusions are seen in the
in other cell types. Their number varies from
perikaryon of neurons. Eat dro-plets are
neuron to neuron and for different parts of the common, as well as Itpochrome or lipofuscliin
same neuron. They are particularly profuse in granules. Some neurons have' pigment
axon terminals (i.e., bulb-like swellings along granule_s,notably melanin and ircon.
or at the ends ofaxons).
Melanin granules are present in nerve'
Lysosomes are also abundant in neurons. cells of the substantia nigra of the midbrain,
Inasmuch as neurons are long-lived, the the locus coeruleus near the fourth ventricle,
lysosomes come in bandy in recycling proteins and the sBinal ami"sympathetic ganglia. Iron
from senescent cellular structures and in dealing granules, .on the other hand, are present in the-
with abnormal and foreign proteins. neurons in the globus pallidus.
Peroxisomes are consistently present in The number of inclusions in neurons,
significant numbers in neurons, where they especially lipofuschin and if-on granules,
are found in the main body, and are smaller noticeahly increases with age.
(about 0.25 to 0.5 urn in diameter) than in
other cells. They probably help in preventing the Cytoskeleton of Neurons
degeneration of the neuron by not allowing the As in other cells,the cytoskeleton of neurons
accumulation of strong oxidizing agents and by is formed by three types of fibrillar elements,
playing a role.in detoxifyingjioxious s~Qstance.s. collectively referred to as neu~ofibrils: 1)
A centrosome (MT0C) is another mierofilaments, 2) intermediate filarrrerrts;
organelle that is generally present in neurons, and 3)microtubules. Neurofibrils are present in
despite the fact that neurons are incapable of cell all neurons but are particularly well-developed
division. It is usually located in the peripheral in large ones. They extend into the axon and the
area of the perikaryon. The cep.trosome of dendrites.
neurons, however, is atypical because it does The microfilaments are the fines of the
not contain centrioles, but it is the source of the fibrillar elements in neurons- Their diameters
microtubules that the cells need. average 5.0 nm. Structurally, they are similar to

Fig. V-2. Neuron. The

photomicrograph shows a
neuron in the spinal cord and
some of its parts including
the nucleus (n), axon (a)which
arises from the axon hillock
(h), and dendrites (d). The
basophilic granules in the
cytoplasm are Nissl granules.
Spinal Cord, H&Ex400.

NERVOUS TISSUE ~
the microfilaments present in other cell types.
Hence, they are made up of the fibrillar type
of actin (F-actin) that consists of two strands
of helically-arranged, polymerized G-actin
filaments.
The intermediate filaments present in
neurons are called neu ofilaments. They are 10
nm in diameter and are present in the cell body
and the cell processes. They are particularly
abundant in the axon. N eurofilaments provide
internal support for the cell and fix the diameter"
of dendrites and axons.
The microtubules in neurons, often referred
to as neurntubiiles, are similar to those found in
other cell types. They provide internal support
for the neurons. They also strengthen synapses
(i.e., the points of contact between neurons
and other cell). Furthermore, they play a role
in the intracellular transport of organelles and
secretory vesicles.
Processes of a Neuron
The processes of a neuron are cytoplasmic
extensions of the cell body. They are often
rather numerous such that the amount of
cytoplasm they contain is more than what is in
the perikaryon. In fact, in most neurons, more
than 90% of the cytoplasm is in the processes.
As stated earlier, there are two types of
processes in neurons: 1) axon which conducts
impulses away from the cell body; and 2)
Fig. V-3. Types of Neurons According to
Number of Processes: a) Unipolar; b) Bipolar; c)
Pseudounipolar; and d) Multipolar.
ESTEBAN& CCNli\LES' TEXTBOOK OF HISTOLOGY
dendrite which carries impulses towards the
cell body. There is one and only one axon per
neuron. Dendrites, on the other hand, could be
absent, solitary, or numerous.
Classification of Neurons
Morphologically, neurons are grouped
into four types based on the' number of their
processes.
1. Unipolar -when only one process, an
axon,·is present. This type exists in early
embryonic life but is rarely present in adults.
2. Rseudounip'olar~ when a Singleprocess,
morphologically an axon, leaves the body,
but soon bifurcate . Neurons of this type
are exemplified by se_nso~¥neurons that
are present in the cFaniosginal ganglia.
3. Bipolar - when a Single dendrite and an
axon arise at opposite poles of the cell body.
Examples of neurons of this type are seen in
the (i)jfaJ:tnr~epithelium of the nose and in
the vestibular and cochleareganglia.
4.,, Multipolar - when numerous dendrites
are present. Most neurons are of this type.
Functionally, neurons are classified into
three types:
1.
CNS.
2. Motor neurons (efferent neurons) which
transmit impulses from the CNS to effector
cells.
3. Interneurons (assodat· on neu-rnnsJ
which convey impulse from one neuron to
another.
Some sensory neurons synapse or
communicate directly with motor neurons.
However, as a rule, sensory stimulus is passed
on by sensory neurons to interneurons which
make up the great majority of neurons in the
nervous system. Interneurons do not simply
relay information. In concert, they also process,
store, and analyze information (stimuli), and
decide on appropriate responses to stimuli.
Dendrite
More than one dendrite is usually present
in a neuron. In fact, m st neurons contain
many dendrites. Dendrites provide most of the
receptive surface of the neuron.
Coverings ofAxons
All axons are enveloped by a sheath of cells,
the neustlermrral sbeafII In addition, many
axons are further encased by a m)':elin slieat ,
and in the PNS, also by a llasallamiIDl'.

In general, dendrites branch more

extensively, but are shorter, than axons.
Dendrites contain Nissl bodies,
mitochondria, and neurofibrils, but they do not
have a Golgi complex.

Schwann cells are encased external to their

Axon (Axis Cylinder)
plasmalemma by basal lamina,» hence, axons in
The axon arises from a conical elevation the PNS are" de-facto" also enveloped by basal
on the perikaryon called the axon:hillock. lamirra,
The axon and the axon hillock, in contrast to
Several Schwann cells are normally needed
dendrites, are devoid ofNissl granules, but the
to completely envelop an axon. The points of
axflplasm (i.e., cytoplasm of the axon) contains
discontinuity between successive Schwann cells
other organelles such as §ER and mitochondria.
are referred to as wdes ot:Ranvie);.
The cell membrane of the perikaryon is
continuous with that of the axon, where it is
called axolemma.
An axon is usually more slender than a
d~~drite but is typically longer. The longest
axons in the human body are those that form
the sciatic nerve which can be longer than one
meter and extend from the inferior end of the
spinal column to the big toe of each foot.
Only one axon is present in a neuron, but it
gives off collateral branches.
An axon forms small, rounded swellings
called houtrrns Cterminals) at the ends th,uIfon
Fig. V-4. Myelinated PNS Nerve Fiber, cross
terminau!X; \te:flnina ontons) or along the
section
course Gbouton en passant~ of its branches. It is
at these axon terminals or boutons that an axon In addition to the Schwann sheath, larger
synapses or communicates with other neurons axons in the peripheral nervous system are
or cells. ' enveloped by a material that is highly refractile
Axons are capable of axona t anspo t, in fresh specimen but black in tissues fixed
meaning substances can move along the with osmium tetroxide. This material is called
axon. Two forms of this transport exist: a) lll~elin. The structure it forms around the axon:
anterograde which involves movement of which lies internal to the Schwann sheath, is
substances from the perikaryon to the axon called my:elin sheath. Thus, there are two kinds
terminals; and b) ~etrograde which involves the ofaxons depending on the presence or absence
transport of substances from the axon terminals of myelin: m:y_elinatea and ~y:elinaied. The
to the perikaryon. myelin sheath has a profound influence on the

NERVOUS TISSUE ..
physiologic properties of a neuron. For one,
the conduction of a nerve impulse is faster in
myelinated than unmyelinated axons.
Schwann cells and myelin were once
considered as separate structures since they
appear distinct from each other when seen
under the light microscope. However, electron
microscopy has proven that myelin is actually
made up of Schwann cell plasma membranes
that have been spirally wrapped, many
times. over, 'ar~und, the axon. In between the
concentrically-arranged plasma membranes,
there is yery little cytoplasm.
- The nodes of Ranvier interrupt the myelin
sheath in the same areas where there are
interruptions in the sheath of Schwann. Thus,
in the nodes of Ranvier, th~ axon is partially
uncovered. Here is where the axon gives off
collateral branches.
In fixed specimens, the layers of myelin in a
myelin sheath may separate in some areas. These
points of separation are referred to 'as incisures
or clefts 0 Schmidt-Lantermann. Under the
electron microscope, the clefts appear to be
shearing defects in the lamellae of the myelin
sheath.
In the central nervous system or eNS, there
are no Schwann cells. Nevertheless, axons still
have neurilemmal sheaths and the larger ones
Fig. V-S. Myelinated eNS Nerve Fiber. Note
that the myelin sheaths of several axons are
formed by a single oligodendrocyte.
ESTEBAN & (iONZJ\LES' TEXTBOOK OF HISTOLOGY
are likewise myelinated. This is because, in the
eNS, the functions of the Schwann cells are
performed by cells called oligodendrocyfes.
However, an oligodendrocyte differs from
a Schwarm cell because the former forms
segments of myelin sheaths of numerous.
neurons while Schwann cells envelop just one
axon each. Furthermore, oligodendrocytes,
unlike Schwarm cells, are not surrounded
by basal lamina. Incidentally, the amount of
cytoplasm associated with myelin in the eNS is
much less than that in the PNS.
As in the PNS, nodes of Ranvier interrupt
the neurilemmal and myelin sheaths ofaxons
in the eNS.
Nerve Fiber
An axon and its coverings (i.e., neurilemmal
sheath), and when present, myelin sheath and
basal lamina comprise a nerve fiber.
In the PNS, every nerve fiber is enveloped
by some amount of connective tissue that is
referred to as endoneurium. In contrast, in the
eNS, nerve fibers are not invested by connective
tissue.
Synapse
A synapse is the point of contact between a
neuron and another neuron or another cell. It is
the site of transmission of a nerve impulse which
can either be excitatory or inhibitory in nature.
It allows neurons to communicate with each
other or with effector (muscle and gland) cells
and accomplish their integration and control
functions.
The total number of synapses that
neurons make in the body is estimated at 1014
or one hundred quadrillion. This gi ves one
an idea of how complex and sophisticated a
communication network the nervous system is.
"
Types of Synapses
Synapses are categorized into two types
according to the mode by which they transmit
an impulse: electrical and chemical.

Fig. V-6. Synapse. The diagram illustrates an
axodendritic synapse.
Electrical Synapses
- filamen s.
Electrical synapses occur rarely. They exist
between some neurons in the brain stem, retina,
and cerebral cortex. Electrical synapses consist
of gap junctions that are similar to those found
in between epithelial and other cells. They
enable neighboring neurons to communicate
with each other by allowing adjacent cells to
exchange molecules and small ions.
Chemical Synapses
Chemical synapses are more common than
electrical synapses. In fact, most synapses that
neurons make are of this type. This is the reason
why the term synapse, unless qualified, refers to
a chemical synapse.
In a (chemical) synapse, the nerve impulse
is transmitted from one neuron to another
cell by means of chemical substances called
neurotransmitters. At present, there are about
SOknown neurotransmitters.
The neuron that communicates the impulse
in a (chemical) synapse is called a presynaptic
neuron. The cell or neuron that receives the
impulse is called a postsymlp1ic cell. The
latter could be a neuron, muscle cell, or a cell
ofagland.
numerous mitochondria, neurofibrils,
lysosomes, and membrane-bound
vesicles (synaptic vesicles) that contain
neurotransmitters. The shape, size, and
contents of synaptic vesicles vary depending
on the function of the presynaptic neuron.
At the synapse, the axolemma of the
presynaptic neuron is thickened and is referred
to as the presInaptic membrane while the
cell membrane of the postsynaptic cell, which
is also thickened, is called the postsynaBtic
membrane. The presynaptic and postsynaptic
membranes are separated by a small gap (20-30
nm) called the sy-napficdeft which may contain
p'olysaccharides and some fine in_te-rsynapfic
The presynaptic membrane, synaptic
cleft, and postsynaptic membrane comprise a
s}'napse.
Impulse Transmission at the Synapse
When an impulse reaches the axon terminal
of a presynaptic neuron, the neurotransmitters
in its synaptic vesicles are released by exocytosis
at the presynaptic membrane into the synaptic
cleft. The neurotransmitters then diffuse
across the synaptic cleft and are taken up by
receptors (proteins) that are at the postsynaptic
membrane. Incidentally, neurotransmitters
have specific receptors.
Synapses between Neurons
Individually, the neurons that comprise
the CNS ar-e not intelligent. But when they
communicate with each other via their synapses,
they are able to collectively process and analyze
information.
Neurons synapse with each other in a
variety of ways. Very commonly, the axon of a
neuron synapses with a dendrite or a perikaryon
of another neuron, forming axodendritic
and axosomatic synapses, respectively.
NERVOUS TISSUE ..
Sometimes, the axon of a neuron synapses
with the axon of another neuron to form
an axoaxonic synapse. Rarely, other
types of contacts occur among neurons-
d e n d r'o d e n d r It i c, somatodendritic,
somatosomatic, somatoaxonic,
dendroaxonic, and axoaxodendritic (serial).
A neuron usually forms multiple synapses
with other neurons. Some neurons carry this
feature to extremes-the Purkinje cells of
the cerebellar cortex, for example, establish
hundreds of thousands of synapses with
neighboring neurons.
Neuroglial Cells (Neuroglia; Glial
cells; Glia)
Interspersed among the neurons in nervous
tissue are supporting cells called neuroglial
cells.Neuroglial cellsprotect neurons; aid them
in performing their functions by creating and
maintaining an appropriate environment where
neurons can carry out their function; and playa
role in neural nutrition.
Neuroglial cellsoutnumber neurons. There
are five to 10 times more neuroglial cells than
neurons in nervous tissue, but they are smaller
than most neurons. Thus, they account for only
half of the volume of nervous tissue.
In the eNS, there are four types of
neuroglial cells:astrocytes, oligodendrocytes,
microglia, and ependymal cells. In the PNS,
there are two: Schwanncells and satellite cells.
Astrocytes and oligodendrocytes are sometimes
collectivelyreferred to as macroglia.
The neuroglial cells, except for the
microglia, arise from embryonic ectoderm.
Microglia evidently arise from embryonic
mesoderm.
Neuroglial cells cannot be distinguished in
H&E preparations. Many times, the nuclei are
the only parts of the cells that can be seen.
Unlike neurons, neuroglial cells have the
capacity to divide by mitosis.
Astrocytes
Astrocytes are the largest and most
abundant of the neuroglial cells. They are star-
shaped and havenumerous,branching processes.
They are involved in many metabolic processes
that occur in nervous tissue. Moreover, they
form scar tissue in damaged areas.
Types of Astrocytes
On the basis of their processes, two types of
astrocytes can be distinguished: protoplasmic
and fibrous.
Protoplasmic astrocytes have abundant
cytoplasm. Their nucleus is bigger and paler-
staining in routine histologic preparations
than that of the other neuroglial cells. They
are found mainly within the gray matter of the
brain and the spinal cord.
Fibrous astrocytes have longer, more
slender processes than protoplasmic astrocytes.
They are located chiefly in the white matter.
Oligodendrocytes (Oligodendroglia)
Oligodendrocytes are smaller, and have
fewer and shorter processes than astrocytes.
They have scanty cytoplasm. Their nucleus,
usually ovoid or spherical, is smaller but
more deeply-staining in routine histologic
preparations than that of astrocytes. They are
located mainly in the white matter of the eNS
where they form the neurilemmal and myelin
sheaths of the axons.
Microglia (Microglial cells)
Microglia, as the name implies, are smaller
than astrocytes and oligodendrocytes. They are
distributed throughout the eNS. Their nuclei
are small and elongated while their cytoplasm
is scanty and contains many lysosomes.
Microglia are phagocytes that remove
cellular debris from sites of injury or normal
cell turnover. They share many properties
with macrophages such that some histologists
categorize them as macrophages even though
I
their lineage is still uncertain. There is, however,
evidence that in the embryo, monocytes from
blood enter the developing brain where they
differentiate into microglia.
Ependymal Cells (Ependymocytes)
Ependymal cells are cuboidal cells that
possess short cilia and microvilli.They also have
cytoplasmic processes on their basal surface
that are relativelyshort except for those present
in some ependymal cellsin the floor ofthe third
ventricle (calledtanycytes), which are very long
and extend into the hypothalamus.
Ependymal cells comprise the simple
cuboidal epithelium that lines the cavities of
the central nervous system (i.e., ventricles of
the brain and the central canal of the spinal
cord) and form the secretory epithelial lining of
the choroid plexuses thaf secrete cerebrospinal
fluid (CSF). Their ciliary movement also helps
circulate CSF.
Schwann Cells
, ,
Schwarm cells form the neurilemmal and
myelin sheaths of peripheral nerves. They have
been discussed earlier in this chapter.
Satellite Cells (Mantle cells;
Amphicytes)
Satellite cells are small, flattened cells that
surround the cell bodies of neurons that are
in ganglia. They are the PNS counterparts of
astrocytes. They provide structural support
for, and are involved in numerous metabolic
processes of neurons.
M cavity and vertebral canal, i.e.,brain and spinal
As mentioned earlier in this chapter,
the nervous tissue in the body is organized
to comprise the nervous system. It is an
extraordinarily complex but highly integrated
communication network that receives,
stores, processes, and sends out voluminous
amounts of information simultaneously and
instantaneously. It is one of the body's two
major integration and control systems-the
other being the endocrine system.
The integration and control functions ofthe
nervous system are performed by 1) collecting
stimuli from the environment by means of
receptors; 2) transmitting these stimuli, called
nerve impulses, to highly organized reception
and correlation areas for interpretation; and 3)
issuing orders to effector organs for appropriate
responses to the stimuli. In contrast, the
endocrine system exerts its influence over cells,
tissues, and organs by producing and releasing
to the blood stream chemical messengers called
hormones.
In general, the response of the nervous
system to stimuli is rapid and precise, but its
effects are brief. In contrast, the response of the
endocrine system to stimuli is slower and more
diffuse, but its effects are longer-lasting.
The nervous and endocrine systems overlap
anatomically and functionally. Their anatomic
overlap is exemplified by the hypothalamus, a
part of the brain that also elaborates hormones.
The nervous system is the organ system
primarily involved with the conscious
experience. This is not to say,.however,that the
functions of the nervous system are all carried
out at the conscious level.
Anatomic Divisions
of the Nervous System
As also previously mentioned, the nervous
system is divided anatomically into two parts:
central nervous system EENS) which refers to
the large mass of nervous tissue in the cranial
cord, respectively; and peripheral nervous
system (PNS) which refers to all other nervous
tissues in the body.
Central Nervous System (CNS)
The CNS has no connective tissue stroma,
hence, the nervous tissue that comprises the
NERVOUS TISSUE ..
brain and the spinal cord is soft and jelly-like.
It is very fragile, thus, it is protected by bony
structures, i.e., skull and vertebral column.
Internal to the bony structures that protect
them, the brain and the spinal cord are further
protected by enveloping membranes called
meninges made up of connective tissue.
Meninges
The meninges that cover the brain and
the spinal cord consist of three layers. The
outermost of these layers is the dura mater
or pachymeninx which is firm and made up
of dense collagenous connective tissue. The
middle layeris the arachnoid membrane while
the innermost membrane that is closelyapplied
to the brain is the pia mater. The two inner
membranes are made up of connective tissue
that is much looser than that in the dura mater.
Fig. V-7. Meninges
They are often considered as a single entity, the
leptomeninx or pia-arachnoid.
In the brain, the outer surface of the
dura mater adheres to the inner aspect of the
cranium. It thus acts as-and is synonymous
with-the periosteum ofthe cranial bones, and
is called periosteal dura.
In the spinal cord, the outer surface of
the dura mater is lined by a simple squamous
epithelium and does not adhere to the vertebrae.
ESTEBAf'-l & GC[\JZ/\LES' TEXTBOOK OF HISTOLOGY
The vertebrae have a distinct periosteum
connected to the dura mater by ligamentous
strands. Thus, a space exists between the
periosteum and dura mater. This space
occupied by fat and venous plexuses is called
epidural space.It has some clinical significance
and is a site for the introduction of some drugs,
including anesthetics (epidural anesthesia).
The inner surface of the dura mater in the
brain and the spinal cord, on the other hand,
which is also lined by a simple squamous
epithelium,is referredto as the meningeal dura.
Between the meningeal dura and the arachnoid
membrane is an area called subdural space.
The subdural space contains minimal amount
of serous fluid and is more of a potential space.
It sometimes becomes clinically significant in
traumatic brain injuriesbecauseblood fromtorn
blood vessels (usually veins) could accumulate
in the space to form a subdural hematoma
that could increase intracranial pressure and
compress and damage brain tissue.
The arachnoid membrane is a flat, sheet-
like membrane that is thinner than the dura
mater. It is smooth on its outer surface, but
projecting from its inner surface are cobweb-
like (thus,the term arachnoid) connective tissue
strands (arachnoid trabeculae) that connect it
to the underlying pia mater.
The pia mater is a thin but highly vascular
loose connective tissue layer that closely
adheres to the substance of the brain and spinal
cord. It spans the entire surface of the brain
and is continuous with the ependyma that lines
the ventricles of the brain. It is separated from
nervous tissue by neuroglial cells. Like the
arachnoid membrane, the pia mater is mainly
made up ofinterlacingbundles of collagenfibers
surrounded by networks of fine elastic fibers.In
between the extracellular fibers,fibroblasts and
macrophages abound.
The arachnoid membrane and the pia mater
are separated by a space, the subarachnoid
space, which contains cerebrospinal fluid
(CSF).

Fig. V-S. Chroid Plexus. H&E x100.
Cerebrospinal Fluid (CSF)
Cerebrospinal fluid is a clear, slightly
viscous fluid that circulates within the ventricles' ,
of the brain, the subarachnoid space, and the
central canal of the spinal cord. Its total amount
in the body is normally 80 to ISO mL. It has
a specific gravity of 1.004-1.008. It contains
sugar, inorganic salts, and traces of protein. The
only cells that are normally present in CSF are
lymphocytes. They are very few, only 1 to 3 per
mL.
CSF protects the central nervous system by
acting as a water cushion. In addition, it plays
an important role in the metabolism of nervous
tissue.
CSF is constantly being renewed. About
500mL of CSF is produced daily, which means
a CSF turnover rate of 3 to 4 times per day. CSF
comes primarily from the choroid plexuses, but
some amount is also produced by the pia mater
and the brain substance.
To keep a fairly constant intracranial
pressure, CSF volume has to be maintained
within normal levels. To accomplish this, some
ventricle
CSF is regularly drained into the venous side
of the circulation via specialized areas of the
arachnoid membrane called arachnoid villi. An
arachnoid villus is a granular structure from the
arachnoid membrane that penetrates the dura
mater and then projects into an intracranial
venous sinus (vein). It acts like a tube with a
one-way valve that allows passage of CSF fr"om
subarachnoid space into the vein, but not of
blood from the vein into the subarachnoid space.
Choroid Plexuses
The choroid plexuses are the chief sources
of CSF. They are located on the roof of the third
and fourth ventricles of the brain and in parts
of the wall of the two lateral ventricles. They
consist of small blood vessels (i.e., arterioles and
capillaries) of the pia mater that form clumps
that protrude into the ventricles. These blood
vessels are lined on their surfaces related to
the ventricles by ependyma. The ependymal
cells that line the choroid plexuses are unlike
ependymal cells elsewhere because the former
form tight junctions with their neighboring
cells. These tight junctions evidently prevent
NERVOUS TISSUE WII

Fig. V-9. Cerebellum. In the eNS, the nerve cell
bodies which comprise the gray matter occupy
the peripheral area while the nerve fibers which
comprise the white matter occupy the central
area. x5.
many substances in blood from becoming part
of eSF. Thus, the ependyma of the choroid
plexuses acts as a blood-CSF barrier.
Arrangement of Neurons in the eNS
The mass of the central nervous system
can be delineated, based on gross coloration,
into two areas: gray matter and white matter.
Gray matter contains the cellbodies, dendrites,
and proximal portions of the axons of the
neurons that populate the eNS and neuroglial
cells. The nuclei of the neurons account for the
color of the gray matter. The rest of the central
nervous system comprises white matter. White
matter does not contain nerve cell bodies, but it
includes those of neuroglial cells in the region.
It also contains the axons of neurons whose cell
bodies are in the gray matter or in a ganglion
(i.e., collection of cell bodies outside the eNS).
The myelin sheath ofthe axons accounts for the
characteristic white color ofwhite matter.
Gray matter occupies the peripheral area
of the brain while white matter occupies the
central area. The reverse is true in the spinal
cord-gray matter is centrally located while
white matter is in the periphery.
In the gray matter, the cell bodies of
neurons with common functions often cluster
FS1TBAN& (JCNZ!\LES TEXTBOOK OF HISTOLOGY
together to form what is called a nucleus (e.g.,
caudate nucleus located in the basal ganglia of
the brain whose neurons are partly responsible
forbody movement and coordination). Regions
of the gray matter where there are numerous
cellbodies not forming distinct nuclei are called
nuclear areas.
In the white matter, on the other hand, nerve
fibershaving a common origin,termination, and
function often bundle together to form what is
known as tract (e.g., lateral spinothalamic tract
in the spinal cord that carries pain, touch and
temperature sensory stimuli to the thalamus
in the brain). Tracts that are flattened are
otherwise calledlemnisci (Singular,lemniscus)
while those that are rounded or thick are called
funiculi (singular, funiculus).
In the eNS, neurons that have long axons
that leave either the eNS or the gray matter and
terminate at some distance in another part ofthe
gray matter are termed Golgi type I neurons.
Neurons that have relatively short axons that
do not leavethe region of the gray matter where
their cell bodies lie are called Golgi type II
neurons.
Peripheral Nervous System (PNS)
Nervous tissue in the PNS is organized in
such a way that the nerve cell bodies are bound
together by some amount of connective tissue
to form ganglia. The nerve fibers are likewise
bound together by connective tissue to form
nerves (peripheral nerves).
The PNS receives and relays all nerve
impulses originating from stimuli from both
within or external to the body to the eNS.
The eNS then integrates these stimuli and
formulates appropriate responses that are then
relayed to the effector cells, tissues, and organs
bythePNS.
Ganglia
A ganglion (plural, ganglia) is a collection
of cell bodies of neurons that have a common
function in the PNS. It isroughlythe counterpart

of a CNS nucleus. Ganglia vary in size. Some
small ones, like those in the myenteric plexus
of Auerbach and submucous plexus (of
Meissner) in the digestive tract, contain only
a few neurons while some large ones contain
thousands.
As a rule, a ganglion is delineated from
surrounding structures by a connective"
tissue capsule. In the ganglion, each neuron is
surrounded by supporting cells called satellite
cells. A neuron and its satellite cells are, in turn,
separated from their neighboring neurons and
their respective satellite cells by connective
tissue elements.
Fig. V-11. Spinal Ganglion. The neurons (n) in
a ganglion are surrounded by supporting cells
called satellite cells (5). H&E x400.
Fig. V-10. Spinal Cord. In the
spinal cord, in contrast to the
eNS, the.white matter occupies
the peripheral region while the
gray matter occupiesthe central
area. x1O.
Nerves (Nerve trunks; Peripheral
nerves)
Nerves are the PNS counterparts of tracts
in the CNS.
A nerve is a collection ofnerve fibersthat are
bunched in groups called bundles or fascicles.
It is enveloped by dense irregular connective
tissue elements called epineurium that keeps
the fasciclestogether.
Within the nerve, each fascicle is
likewise encased by connective tissue called
perineurium. The perineurium keeps the
nerve fibers within the fascicletogether.
Within the fascicle,each of the nerve fibers
is also individually wrapped and supported
by delicate, loose connective tissue called
endoneurium.
Nerves whose cellbodies are in the brain are
called cranial nerves while nerves whose cell
bodies are in the spinal cord are called spinal
nerves. There are 12pairs of cranial nerves and
31pairs of spinal nerves. Incidentally, one of the
cranial nerves, eN II (optic nerve) is not really
a nerve because it does not leave the brain. It is
actually a tract.
The cranial and spinal nerves, as well as
the nerves whose cell bodies are in the ganglia,
combine, interconnect, and branch off in
various ways to form the numerous named and
NERVOUS TISSUE __
Fig. V-12. Nerve, cross section. Note
the epineurium that envelops the whole
nerve and the perineurium that encloses
the fascicles. Masson's trichrome x100.

unnamed nerves that end in the different tissues
and organs ofthe body. Among the largernamed
nerves are the ulnar nerve in the forearm and
sciatic nerve in the lower extremity.
Functionally, most nerves are mixed
nerves, i.e., they contain both afferent
(sensory) and efferent (motor or secretory)
fibers. Afferent nerve fibers contain the axons
of sensory (afferent) neurons. They transmit
impulses from the skin, muscles,bones.Internal
organs, and special senses to the CNS. Efferent
nerve fibers contain the axons of efferent
(motor) neurons that order muscles to contract
and glands to secrete.
Nerve Endings
Simple (Free) and Expanded-tip Nerve
Endings
Simple nerve endings are merely the naked
(i.e., devoid ofneurilemmal and myelin sheaths)
terminations ofaxons of afferent nerves. They
are found in all tissues. They can discern
pressure but are most sensitive to touch, pain,
and temperature.
At this point, some confusion may occur in
the mind ofthe student on how a nerve, which is
made up ofaxons and their coverings, can serve
a sensory function when axons are supposed to
transmit stimulus awayfrom the cell body and,
therefore, should only be motor in function.
Clarification about this matter is thus in order.

The terminations of nerves in epithelial, The sensory neurons are pseudounipolar

connective, and muscle tissues are called nerve neurons whose cell bodies are in the
endings. The terminations of afferent nerves craniospinal ganglia (encephalospinal
are called sensory (afferent) nerve endings ganglia), i.e., sensory ganglia on the dorsal
root of spinal nerves and ganglia of cranial
while those of efferent nerves are referred to as
motor (efferent) nerve endings. nerves that contain general sensory and taste
fibers. Being pseudounipolar, they have a single
Sensory Nerve Endings (Sensory Nerve process, an axon, which bifurcates into two
Receptors) unequal-but both myelinated-branches
Sensory nerve endings collect stimuli and a short distance from the perikaryon. The
are dispersed all overthe body.Morphologically, smaller branch is poorly-myelinated and is
they are categorized into three groups: simple functionally and morphologically an axonwhile
(free), expanded-tip, and encapsulated. the larger branch is functionally a dendrite.

ESTES/'»") 8" GO\iz/\L.ES' TEXTBOOK OF HISTOLOGY

r--

Fig. V-13. Smooth Muscle, cross section. Note that in the cells where the nucleus is seen, the nucleus is
centrally located. At tip of arrow is endomysium that envelops each muscle cell. Duodenum, H&Ex400.

However,because the latter is well-myelinated, A Ruffini's corpuscle is a small, spindle-

it is also generally called an axon or sensory shaped structure seen in the dermis of the skin,
axon. tendons, and ligaments. It consists of bulb-like
expansions of the terminal branches of a naked
Expanded-tip endings are exemplified by
axon that are enclosedby avery thin connective
Merkel discs found in the skin and mucosal,
tissue capsule. It is sensitive to deep pressure
surfaces. A Merkel disc consists of the naked and stretch.
leaf-like terminal of an axon that is in contact
End bulbs of Krause are found in the
with a Merkel cell. The latter is a modified
conjunctiva and mucous membrane of the
epithelial cell as described in the chapter on
lips, dermis, glans penis, and clitoris. They are
the skin and its appendages. As a rule, a single
afferent nerve fiber makes contact with many
Merkel cells.Merkel discs are sensitiveto touch
and pressure.

Encapsulated Nerve Endings

Encapsulated nerve endings are made
up of naked axon terminals enclosed by a
lamellated connective tissue capsule. There
are many kinds of encapsulated nerve endings,
some of the more common ones are Ruffini's
corpuscle, end bulb of Krause, Vater-
Pacinian corpuscle, Meissner's corpuscle,
neuromuscular spindles (muscle spindles),
and Golgi tendon organs. The last two are
associated with muscle tissue and are discussed Fig. V-14. Vater-Pacinian Corpuscle (at arrows).
in the chapter on muscle tissue. Hypodermis, H&E x400.

NERVOUS TISSUE va
 Fig. V-1S. Meissner's Corpuscle (at arrows). Skin, H&E x400.
minute (SOurn) oval structures. They consist
of an axon enclosed by a thin, lamellated
capsule consisting of connective tissue. The
axon typically arborizes within the capsule and
its terminal branches intertwine. End bulbs
of Krause are probably tactile and pressure
receptors.
The Vater-Eacinian corpuscle is the largest
. of the sensory nerve endings. It is a white,
oval structure that can reach a diameter of 0.5
mm and length of up to 2 cm. It is sometimes
appreciated with the naked eye. In routine
histologic preparations, it looks like the cut
surface of an onion. The capsule of the Vater-
Pacinian corpuscle consists of30 or more layers
of circularly-arranged flattened cells (probably
modified Schwarm cells). The corpuscle is
usually supplied with a single axon. In the
capsule, the axon gives off numerous bulbous
terminal branches. Vater-Pacinian corpuscles
are widely distributed in the body and can
easily be found in the dermis, subcutaneous
connective tissue, pancreas, mammary glands,
ESTEB;\f\j & c)Of\jZi\L.ES' TEXTBOOK OF HISTOLOGY
the mesenteries,and the external genitalia.They
ate sensitive to vibration, stretch, and pressure
(coarse touch).
The Meissner's corpuscle is seen in the
dermis of the skin of the fingers, toes, palms,
and soles. It is smaller than a Vater-Pacinian
corpuscle. It is a cylindrical structure whose
long axis is perpendicular to the skin surface.
It has a capsule that encloses a mass of ovoid
cellsthat are arranged perpendicular to the long
axis of the corpuscle. The axon that supplies
the corpuscle enters the capsule at its inferior
pole. Inside the capsule, the axon follows a
tortuous route and ends in the superior pole of
the corpuscle.
The Meissner's corpuscle is a tactile (touch)
receptor.
Motor Nerve Endings
Motor nerve endings are responsible for
transmitting the stimulus that commands
muscle fibers to contract and glandular cells to
secrete.
 The axon terminals of the efferent nerve
fibers of the somatic motor neurons (i.e.,
motor neurons that supply skeletal muscles) and
the skeletal muscle fibers that they innervate
forrn specialized junctions called motor
endplates (discussed in the chapter on muscle
tissue).
In contrast, the axon terminals of the
efferent nerve fibers of the visceral motor
neurons (i.e., motor neurons that supply cardiac
and smooth muscles fibers and glandular cells)
do not form specialized junctional complexes
with the cells that they innervate. They simply
end at a short distance from the muscle fibers
and the glandular cells that they innervate.
Functional Divisions of the Nervous
System
The nervous system is anatomically divided
into the eNS and PNS. Functionally, however,
it is divided into the somatic nervous system
(SNS) and autonomic nervous system (ANS).
The somaticnervous system (SNS)includes
all neurons in the eNS and PNS associatedwith
muscles, skin, and sense organs regardless of
whether they are afferent (sensory), efferent
(motor) , or interneurons (association). The
autonomic nervous system (ANS), on the other
hand, is composed of all neurons in the eNS and
PNS which are concerned with the regulation.
of visceral organs also regardless of whether
they are afferent (sensory), efferent (motor), or
interneurons (association).
Somatic Nervous System (SNS)
Somatic afferent (sensory) neurons are
responsible for the reception of sensory stimuli
from the external environment (e.g.,touch and
temperature) and proprioceptive stimuli
from skeletal muscles, tendons, and joints.
Their terminations have been described in the
preceding section.
Somatic efferent (motor) neurons, on
the other hand, innervate the skeletal muscles
responsible for voluntary movements of the
body,The terminations oftheir axons (discussed
in the chapter on muscle tissue) participate in
the formation of the motor end plates on the
surface of the muscle fibers they innervate.
Autonomic Nervous, System (ANS)
Traditionally, the ANS has been defined
as consisting only of visceral efferent (motor)
neurons; the visceral afferent (visceral
sensory; autonomic afferent) neurons-are
grouped with the SNS.This is probablybecause,
morphologically, the receptors of the visceral
of tPL¬
afferent neurons- are similar to- tP.0:0-&-2
somatic afferent neurons, except that they are
associated with smooth muscles and glands,
and their fibers run for a part of their course
with the autonomic efferent fibers. Authors are
now increasinglygrouping the visceral afferent
neurons with the ANS.
The visceral efferent neurons control
the activity of cardiac and smooth muscles,
and glands-structures that are not under
conscious control. They differ from the somatic
efferent neurons because two visceral efferent
neurons, instead of only one, are involvedin the
transmission of an impulse from the eNS to the
effector cells. The cell body of the first visceral
efferent neuron (preganglionic neuron) is
in the eNS while the cell body of the second
visceral efferent_neuron (postganglionic
neuron) is in an autonomic ganglion. The
axon (preganglionic fiber) ofthe preganglionic
neuron leavesthe eNS and enters an autonomic
ganglion to synapse with the postganglionic
neuron. The axon (postganglionic fiber) of
the postganglionic neuron, on the other hand,
leaves the autonomic ganglion to terminate in
an effector organ.
Preganglionic fibers leave the central
nervous system at severallevels,via the: 1) III,
VII, IX, and X cranial nerves; 2) thoracic
spinal nerves; 3) upper lumbar spinal nerves;
and 4) sacral spinal nerves.
NERVOUS TISSUE ...
Divisions of the Autonomic Nervous
System (ANS)
The neurons that comprise the visceral
efferent branch of the ANS are grouped into
three categories or divisions: sympathetic
(thoracolumbar), parasympathetic
(craniosacral), and enteric.
• Sympathetic Division of the ANS
The sympathetic division (sympathetic
nervous system) of the ANS consists of 1)
preganglionic neurons whose fibers exit the
CNS via the thoracic and lumbar spinal
nerves; and 2) the postganglionic neurons
in the vertebral and prevertebral ganglia
with which the fibers of preganglionic
neurons synapse. Incidentally, the vertebral
ganglia are collectively referred to as the
sympathetic trunk.
The sympathetic nervous system
responds to impending danger or stress.
It is responsible for the increase of one's
heartbeat and blood pressure, sense of
excitement, and other physiological changes
that occur in "fight or flight" situations.
• Parasympathetic Division of the ANS
The parasympathetic division
(parasympathetic nervous system) of
the ANS, on the other hand, refers to 1) the
preganglionic neurons whose fibers leave
ESTES!\i-.J
& C()NZ/\LES' TEXTBOOK OF HISTOLOGY
the CNS via the cranial and sacral spinal
nerves; and 2) the postganglionic neurons in
ganglia (which are near or within the walls
of the structures they innervate) with which
the fibers of preganglionic neurons connect.
The parasympathetic nervous system
is called upon during resting and relaxing
situations. It is responsible for things such
as constriction of the pupil, slowing of heart
rate, and dilation of the blood vessels.
• Enteric Division of the ANS
The enteric division (enteric nervous
system) of the ANS, meanwhile, is made
up of the visceral efferent neurons whose
cell bodies and fibers form ganglionated
plexuses (i.e., ganglions that interconnect)
in the walls ofthe digestive tract as well as in
the pancreas and the gallbladder.The enteric
nervous system has connections with the
sympathetic and parasympathetic nervous
systems,but it functions autonomously.Even
when its connections with the sympathetic
..and parasympathetic nervous systems are
severed,it still is ableto carry out its assigned
function to regulate the activities of the
muscles and glands of the digestive tract.
The enteric nervous system is
responsible for regulating the activities of
/
the digestive tract.
 Chapter VI

Cartilage
and Bone . ,

he adult human skeleton which serves in the kind and amount of extracellular fibers

T as the framework of the body consists

of 206 named bones (e.g., sternum or
breast bone), a few unnamed sesamoid bones,
they contain.
There are no blood vessels, lymph vessels,
or nerves in cartilage. The chondrocytes are
and numerous cartilages, some of which are also nourished by diffusion of nutrients from the
named. These bones and cartilages are actually surrounding tissues.
organs because although they are mainly made
up ofbone and cartilage tissue, respectively, they
Hyaline Cartilage
have associated blood vessels and are invested
and/or lined by connective tissue. Thus, the Hyaline cartilage is the most abundant type
terms cartilage and bone, like the term muscle, of cartilage in the body. It is glistening, smooth,
can refer either to a tissue or an organ. and pearly white in fresh specimens.
Cartilage and bone tissues are special Hyaline cartilage forms the bulk of the
types of dense regular connective tissue. Like skeleton of the fetus in-utero. It is ideal for
any dense regular connective tissue, they have this purpose because besides being strong
relatively few cells and abundant intercellular enough to serve as a supporting framework for
substance. However, cartilage and bone differ the body of the fetus, it also has the capability
from dense regular connective tissue because to grow rapidly in conditions of relatively low
their cells, called chondrocy:tesand osteoc}':teSl, oxygen tension. Most of the bones of the xial
respectively, are inside cavities called Iacunaea ami a12l1endicularskeletoQjare first formed out
Their intercellula); substam:e (matrix) is also of hyaline cartilage. By the secnrrd month of
firmer or harder. The inte1'Lellularsubs ance ob intrauterLne life, however, the cartilages that
cartilage is gelatinous, albeit firm, while that of comprise the fetal skeleton start to get replaced
hone is calcified, hence, rigid and hard. by bone. This process continues postnatally
such that in adults, hyaline cartilages exist only
CARTILAGE (GRISTLE) in limited areas in and around joiats, the sterna
ends of ribs, some parts of the respiratori)j
There are three types of cartilage: hyaline, system and extern:al ear, and tendons and
elastic, and fibrous. These types differ mainly ligaments that are pressed on by bone.
 /
Fig. VI-1. Hyaline Cartilage. The section shows
part of the perichondrium (p): mature chondrocytes
(ce)inside lacunae(I) and young chondrocytes (cs).
H&E x100.
Composition and Microscopic
Structure of Hyaline Cartilage Tissue
. Hyaline cartilage consists of cells
(chondrocytes) that are inside cavities
(lacunae) which are embedded in the in the
intercellular substance (cartilage matrix).
Chondrocyte (Cartilage Cell)
In vivo, a chondrocy:t~ fills the lacunae
where it resides. However, in routine histologic
preparations, the cytoplasm of the cell usually
shrinks a little.
The qcfoplasm of cho__ndr.oc:y:tesis finely
granular and basophilic. It contains a limited
number of mitochondria. Consistent with the
ESTEBAN & CcmZALES' TEXTBOOK OF HISTOLOGY
./
Fig. VI-2. Hyaline Cartilage. This section, taken
under high power magnification, shows nuclei
(nu) of the chondrocytes (ce) inside the lacunae
(la). The chondrocytes that are clustered together
are isogenous cells (ic). The cartilage matrix
immediately surrounding the lacunae is more
basophilic and is referred to as territorial matrix
(tm). The rest of the cartilage matrix is referred to
as interterritorial matrix (im). H&E x400.
secretory character of the cell, it has a well-
developed rER and Golgi complex. It also
contains inclusions such as fat droplets and
glycogen granules. Chondrocytes possess
cytoplasmic processes though these may not be
apparent in H&E prepar-ations.
or more nucleoli.
are capable of
mitosis, but mature ones are not. When young
chondrocytes mitose, the daughter cells, called
isoge_Dous cells, tend to stay close to each
other. Sometimes, under the light microscope,
isogenous cells may appear to occupy the same
lacuna. As a rule, however, they are separated
from each other by a thin layer of intercellular
substance.
Cartilage Matrix
Hyaline cartilage matrix consists of
an amorphous ground substance where
extracellular fibers are embedded.
Apart from water which accounts for
70% to 80% of the wet weight of the tissue} the
ground substance of hyaline cartilage is mostly
made up of~roteogly:cans. These are organized
in a manner similar to that which prevails in
connective tissue proper.
The G:A:fis} because of their sulfate}
carboxyl} and hydroxyl groups} are strongly
acidic. This is the reason why the irrterrelltrlar
substance of hy-aline 'cartHage is intensely
basophilic.
The in ter c el lu la r substance that
immediately surrounds a lacuna is particularly
rich in GAGs. Hence} this area is more
basophilic than the rest of the matrix. Early
histologists referred to this area} which may
be as much as SO ~m in thickness} as capsule
<of the cnonarocytes. It is now more often
called territorial matrix. Meanwhile} the
area in between the terr itor ial matrices is
called Interrerrftorfal matrix. Structurally}
the innermost (1-3 urn) layer of the territorial
matrix resembles a basal lamina and is
sometimes called pericellular capsule. It
probably protects the chondrocyte against
mechanical stress.
Extr:acellular F-il2)ers
The extracellular fibers in hyaline cartilage
are ty:pe:n collagen fibers. They account for
about 40% of the dry weiglit of the tissue. They
are thinner than those in connective tissue
proper (type I collagen).
Fig. VI-3. Elastic Cartilage. The section is
especially stained to demonstrate the elastic fibers
(black streaks) in the matrix. The nuclei of many of
the chondrocytes can be appreciated. Orcein x200.
Often} in H&E preparations} the collagen
fibers in hyaline cartilage are not distinguishable
because they have the same refractive index as
the ground substance.
Elastic Cartilage
rElastic cartilage is more flexible than
hyaline cartilage and is yellowish in fresh
specimens. It is present in the auricle and
<external acoustic meatus of the ear} auditory:
tube} epiglottis} and some other parts of the
larynx.
Elastic cartilage is morphologically similar
to hyaline cartilage except that the former's
matrix is less abundant. In addition to collagen
fibers that are likewise made up of collage..n,
type II} it also contains a considerable quantity
of elastic fibers. These fibers account for the
pliability and yellowish color of the tissue when
viewed with the naked eye.
Fibrous Cartilage
Eiorous cartilage can withstand greater
stress than hyaline or elastic cartilage. It is
white in fresh specimens. It is the cartilage
d'scs,
type that makes up the int:e.r:ve.rte...b.ral
articular iliscs, and glenoid and acda6ulali
CARTILAGE AND BONE elM
 _ells,which can transform into cliondroblasts
under the right conditions.
Chondroblasts are cells that synthesize the
precursors of the extracellular fibers and the
other organic constituents of cartilage matrix.
When they get surrounded by the matrix they
secreted, they acquire lacunae and transform
into chundrocytes. ~ung chondroc:y.tes
retain their capacity to mitose and synthesize
components ofthe extracellular matrix. Because
of this ability, many authors still call them
Fig. VI-4. Fibrocartilage. Note the abundant
amount of collagen fibers that surround the
chondroblasts despite the fact that they are
chondrocytes. H&E x400. already inside lacunae.
In fibrous cartilage, the perichondrium
labra. Fibrous cartilage can also be found in cannot be appreciated as a distinct entity
surface layers of tendons and trgamenrs that because it blends with the dense regular
are pressed on by bone. connective tissue that invariably surrounds the
tissue.
The ~nllagen fttters of fibrous cartilage are
thickerthan those in hyalineand elasticcartilages
because their collagen content IS type I. BONE
Fibrous cartilage can be regarded as a
articular
transitional stage between dense regular cartilage epiphyseal
plate
connective tissue and cartilage. Under the light
sptlngy. ~ proximal
microscope, it could be mistaken for dense bone ~ epiphysi;
connective tissue because it has an abundance spongy
of collagen fibers-that make the tissue bone·
acidophilic-and a relative paucity of ground medullary
~avjty
substance. However, closer scrutiny shows that
compact
the cells (chondrocytes) are inside lacunae and bone
bigger than fibroblasts.
diaphYSis

Perichondrium :periosteum

With afewexceptions,notablythe a_rticular

cartilage--s, all cartilages are enveloped bJ
dense irregular connective tissue. This tissue
is special because it has chondrogenic potency
(i.e.,ability to form cartilage). This special type distal
epiphysis
of connective tissue, called I!erichondrium,
consists of an outer layer Efillrous la}"eJi)that Fig. VI-S. Parts of a Typical Long Bone
blends with the surrounding tissue', and an
inner, more cellular layer (clionilrogenic layer) The bones that comprise the bulk of the
that adheres to the cartilage. The cells of the adult skeleton form a rigid framework for the
chondrogenic layerapposed to the surface ofthe body. They also protect vital organs (e.g., the
cartilage are stem cells, called,osleop- ogenitor cranium protects the brain whilethe bony thorax

ESTEBANs GONZALES' TEXTBOOK OF HISTOLOGY

 protects the heart, lungs, and other mediastinal
structures). In addition, bones serve as levers
for muscles, and as storehouses for calcium and patella or knee cap which is rather large, most
phosphorus. Likewise, they house bone marrow sesamoid bones are in the form of unnamed
where most of the formed elements of blood are small nodules.
produced.

Forms of Bone Tissue

Types of Bones According to
Shape Two forms of bone tissue are grossly
distinguishable-compact Ecortical, dense)
Grossly,the bones of the body are classified
and 'spongy (cancellous). To the naked eye,
according to shape. The classes include long,
compact none appears like a solid mass while
short, flat, irregular, and sesamoid bones.
spong¥ none has numerous spaces or cavities.

- bong bones are confined to the extremities.

A typical long bone is tubular and consists of
a bo_dy or sha£t and two ends Enroximal and
All the bones of the body consist of an
inner region that is made up of <spongy-: bone
distal ep'ip'hyses~. The shaft has a hollow and an outer casing of compact Done.The only
COI:e'I{meaullar:y: ca~dty) while the epiphyses exceptions are those bones where the central
articulate with other bones and are covered on area is occupied by ai sinuses (e.g.,some bones
their articulating surfaces by hyaline cartilage of the face which contain paranasal sinuses) or
tarticulare caFtilage). by a edullarx caY'ty(e.g.,long bones). In these
latter bones, spongy bone may be minimal in
amount or even entirely absent.
f.la nones are typified by the sternum~' In terms of volume, there is more spongy
scapulae, and many bones of the skull. bone than compact bone in the body: In terms
Donesinclude the veIteJ>rae, IH'll of mass, spa g bone accounts for only 20%
lr_r_egular
=':
Ilones, and the bones of the skull that are not to 25% of the body's total skeletal mass while
flat. (compactnone accounts for 75%to 80%.

Fig. VI-6. Spongy Bone. This

form of bone consists of bone
fragments or spicules (bs) that
are embedded in the bone
marrow (bm) which fills the
marrow cavity. Adipose tissue
(at) occupies a considerable
area of the bone marrow. H&E
x100.

CARTILAGE AND BONE ~

Periosteum and Endosteum
All the surfaces ofbones, except articulating
surfaces and surfaces where muscles, tendons,
and ligaments are attached, are lined or covered
by dense irregular connective tissue. This kind
of tissue is special because it has osteogenic
potency (i.e., ability to form bone). This special
connective tissue is called periosteum ifit covers
the external surface of bone or e_ndosteu:mif it
lines an internal surface or cavity in bone.
Periosteum
Periosteum consists of an outer layer
~fiDrousJay:er)that blends with the surrounding
tissue, and an inner, more cellular layer
~teogenic lay;er) that adheres to the bone.
The cells of the osteogenic layer that abut the
surface of the bone are osteoprogenito ceUs.
Sometimes, bundles of collagen fibers from
the periosteum get trapped within the bone
matrix and form distinct structural entities
called 8harpe¥'s fi ers which can be seen in
light microscopic preparations. Sharpey's fibers
serve to firmly anchor the periosteum to the
bone.
Endosteum
Endosteum lines all the medullary, marrow,
and vascular cavities of bones. It is considerably
thinner than periosteum and often consists
simply of a single layer of osteoprogenitor cells.
Composition, Microscopic
Structure, and Architecture
of Bone Tissue
In both spongy and compact bones, the
.ntercellular substance (bone matrix), which is
very hard, is arranged in thin layers ~lamellae)
that are about 3 to 7 ~m thick each. Dispersed
uniformly in the bone lamellae are cavities
(lacunaeJ, from which numerous small canals
~canalicul~jl) radiate. The canaliculi of each
lacuna anastomose freely with the canaliculi
of surrounding lacunae. A lacun is occupied
ESTES;.\N & CiONZALES TEXTBOOK OF HISTOLOGY
by a cell (osteocyte). Osteocytes have
cytoplasmic processes in the canaliculi. Within
the canaliculi, the cytoplasmic processes of
neighboring osteocytes are in contact with each
other.
Architecture of Spongx Bone
Spongy bone consists of numerous small
interconnecting bone frag_roe_nts Esnicule.~j
traBecula ~ that form the framework of a
very complex system of spaces ~bone marrow
cavities). Each bone spicule is composed
of several parallel bone lamellae and their
associated lacunae and osteocytes. The bone
marrow cavities, on the other hand, are occupied
by bone marrow.
In spongy bone, the osteocytes derive
nutrients and oxygen, either directly or
indirectly, from the blood vessels in the bone
marrow. Those closest to the marrow cavity
obtain nutrients and oxygen directly from the
blood vessels in the bone marrow. Those that
do not have direct access to the marrow cavity
receive sustenance through the connections that
their processes make (within the canaliculi)
with the processes of the cells closer to the
marrow.
Architecture of Compact Bone
t-,,;;. •
olnll
Fig. VI-7. Compact Bone. The diagram shows the
relationship between Haversian canals, Volkmann's
canals, and lacunae.
 Fig. VI-B. Compact Bone,
longitudinal section. Thespecimen
shows a Volkmann's canal (Vc)that
connects a Haversian canal (Hc)
to the periosteum (Pe). Note
that the lacunae in the Havesian
system (Hs) that abut on the
periosteum are oriented parallel
to the Haversiancanal. H&E x100.

---

Fig. VI-9. Compact Bone,

longitudinal section. Several
Haversian systems (Hs) comprise
the section. The Haversian canals
(Hc) are connected to each other
and to the periosteum (pe) by
Volkmann's canals(Vc). H&E x100.

In compact bone, instead of forming oounoaq(; is often delineated by a cement line

spicules,the lamellaeare organized and arranged made up of a thin layer of mineralized matrix
in any of three ways: Haversian systems, and collagen fibers.
interstitial lamellae, or circumferential The Haversian canals of neighboring
lamellae. Haversian systems are connected to each other
Maye_rsian_s}':sJe..ms
or osteons make up the by transverse channels called iYolbn--ann's
bulk of compact bones. A Haversian system caRals. Like Haversian canals, Volkmann's
consists of several bone lamellae-together canals contain blood vessels and nerves and are
with their associated lacunae and osteocytes- lined by endosteum.
that are arranged concentrically around a tiny The blood vessels in the Volkmann's canals
(microscopic) endosteum-lined, longitudinal interconnect with the blood vessels in' the
channel ~Haversian canalj canal of Havers~ Haversian canals and the blood vessels in the
that contains blood vessels and nerves. Its outen periosteum, medullary, and marrow cavities.

CARTILAGE AND BONE CiiI

Fig. VI-10. Compact Bone, cross
section. Compact bone consists
mainly of Haversian systems (Hs)
made up of bony lamellae arranged
around a Haversian canal (He).
Many interstitial lamellae (il) are also
shown in the section. H&E x 100.

FLg. VI-11. Haversian System.

The "section demonstrates the
components 6f a Haversian system, "-
delineated from the ones around
it by cement lines (cI).· At the
center of the system is a Haversian
canal (He). Arranged concentrically
around the canal are bony lamellae
where lacunae (lei) that contain
osteocyte's (os) are seen. Canaliculi
(ca),appearing asdark lines, radiate
from the lacunae. H&E x400.

With these connections, the blood vessels in
the Haversian and Volkmann's canals, together
with the blood vessels in the periosteum and
medullary cavity, are able to deliver nutrients,
oxygen, and other needed substance . They
can also remove metabolic waste products and
carbon dioxide from all the osteocytes in all the
Haversian systems.
In between Haversian systems, bone
lamellae that are not arranged around a
Haversian canal are frequently present. These
pieces of bone tissues are called inferstitial
lamellae. Some authors believe that these are
remnants of Haversian systems that are being
resorbed or broken down (see discussion on
bone remodeling).
CircULUferenti~a~l~~~==_
category of lamellar arrangements in compact
bone, are most developed in long bones. In the
diaphysis of these bones, immediately beneath
the periosteum, there are usually several
lamellae that encircle the whole bone. These
lamellae. In these same bones, the inner aspect
of the compact bone is marked off by lamellae
that encircle the whole medullary cavity.These
lamellae form the
lameltae.

ESTEBAN& G()NZf\LES' TEXTBOOK OF HISTOLOGY

Bone Matrix
Bone matrix, like cartilage matrix, consists
of an amorphous ground substance where
extracellular fibers are embedded.
Ground Substance of Bone
The ground substance of bone consists
mainly of water, but its structural components
are made up of inorganic-minerals and some
or anic substances. he inorganic minerals
account for about two-thirds of the dry weight
of bone while the organic constituents account
for the remaining third.
Inorganic Minerals in the Ground
Substance of Bone
The inorganic minerals in the intercellular
substance of bone consists mainly of calciu
and 12hospJiQus. These form crystals very embryonicmesenchy:mal celts. They are stem
similar to those of cak· m liyaroxyapatite
(€alotP04~6t{)H)J In electron micrographs, multiply infinitely but they can differentiate
these crystals are seen immersed in the ground only into either osteobla_sts or-cho__odroblasts.
substance and lying at regular intervals
alongside the collagen fibers. . . periosteum and the endosteum that directly
The other inorganic substances in bone
include bicar::nonate, citrate,
Organic Substances in the Ground
Substance of Bone
Just like in cartilage, the ground substance
of bone contains prctenglycan molecules, but
smaller than those found in cartilage. Also, the
"core 12roteins"61 toe proteoglycan molecules
are shorter than those in cartilage, and fewer
gly:cosaminoglycans CGltGs9are attached to
them.
The ground substance of bone contains
as well some non-collagenic proteins and
glycoproteins, a few of which are unique to
bone. Some histologists are of the opinion that
the glycoproteins uniquely found in bone are
responsible for the calcification of bone matrix
because connective tissue that has none ofthese
glycoproteins does not calcify.
Extracellular Fibers in Bone
. The extracellular fibers, which .make
up about 90% of the organic content of bone
matrix, are mostly type I collagen fibers. They
are so abundant that they .render borie matrix
acidophilic despite the presence of GAGs.-
Cells of Bone
Four types of cells are present in
bone: osteoprogenrtor cells, osteoblasts,
osteocytes, and osteoclasts. The first three
have the samelineagewhile osteoclasts originate
from cells in the bone marrow.
Osteoprogenitor Cell (Osteogenic_
Cell)
esteoprogenitor cells differentiate from
cells that have a limited potential. They can
In bone, they form the layer of cells of the
abuts the bone. Similarly, in cartilage, they
comprise the layer of cells of the perichondrium
that directly adjoins the cartilage.
Osteoprogenitor cells are fusiform cells.
They cannot be differentiated from fibroblasts
or mesenchymal cells with certainty in routine
histologic preparations.
Osteoblast
(Jsteoblasts, as mentioned above,
differentiate from osteopLogenifof cells.
They are responsible for the synthesis of the
precursors of collagen fibers as well as the
other organic constituents of bone matrix. In
histologic sections, they are easiest to find on
the surface of developing bones and in tissues
where new bone is being formed.
When actively synthesizing secretory
materials, an osteoblast is a relatively large cell
that is round and polygonal or cuboidal in shape.
It has numerous cytoplasmic processes that
CARTILAGE AND BONE '&

Fig. VI-12. Osteocytes and Osteoblasts. Most
of the osteoblasts-which are highly basophilic-
are on the surface of the bone spicules while the
osteocytes are inside lacunae. Spongy Bone, H&E
x400.
are in contact with the cytoplasmic processes
of neighboring osteoblasts. Its cytoplasm is
intensely basophilic, owing to an abundance of
rER.1t also has a well-developed Golgi complex.
This explains why a negative Golgi image is
often appreciated in osteoblasts under the light
microscope. Its single nucleus is situated in the
part of the ~ell that is farthest from the bone
surface.
Osteoblasts lay down their secretion
around themselves. In the process, they get
completely immersed in bone matrix. When this
matrix calcifies, they get trapped and transform
into osteocytes.
In addition to synthesizing the precursors
of collagen fibers and other components ofbone
matrix, osteoblasts also produce growth factors
that promote bone growth.
Recent research data also suggest that
osteoblasts aid osteoclasts in bone resor};'!tion.
Osteoblasts perform this function by at least
two ways. Firstly, they secrete enzymes that
remove uncalcified bone tissue, which evidently
prevents osteoclasts from resorbing the bone.
Secondly, under the influence of parathyroid
IinrIDon'e GBnathormorte), they secrete a
hormone Eosteodast stimulating factor) that
promotes increased activity of osteoclasts.
ESTEBAN& CiCNU"LES' TEXTBOOK OF HISTOLOGY
When osteoblasts are inactive, such as
when there is no need to form new bone,
they lose their basophilia, flatten out, and
become morphologically indistinguishable
from osteoprogenitor cells. But unlike
osteoprogenitor cells, they can neither divide
nor differentiate into other cell types. They can
only revert to being active osteoblasts.
Osteocyte
est'eocxt'eSJ are the cells that occupy the
lacunae in bone tissue. As a rule, only one
osteocyte resides in a lacuna, but occasionally,
more than one may be present. A layer of
osteoid tissue (i.e., uncalcified bone matrix)
usually separates the osteocyte within a lacuna
from the calcified matrix. However, this layer is
so thin that it is distinguishable only in electron
micrographs.
In H&E preparations of decalcified bone,
the morphology of osteocytes is difficult to
appreciate. In special preparations though,
osteocytes are seen as flat cells with numerous
'cytoplasmic processes that occupy the
canaliculi of the lacunae. The light microscopic
appearances of the nucleus and the cytoplasmic
organelles of the osteocytes are similar to those
of inactive osteoblasts.
Within the canaliculi, the processes of
osteocytes are in contact with the processes of
neighhoring osteocytes by means of side-to-
side junctions, probably of the gap type. Thus,
substances are able to pass from osteocyte to
osteocyte via these side-to-side junctions.
Osteocytes do not divide but they retain
enough secretory capability to maintain the
bone matrix that surrounds them. They were
formerly believed to be capable of osteolysis
(i.e., dissolution of bone), and can thus, under
the influence of parathormone, ~ze SOffle
affiouflt of ealeiuM from the bone matrix that
surrounds them. This concept is no longer
universally accepted because recent evidence
points to the osteoblast as the only cell type in
bone that has receptors for parathormone.
 In the process of bone resorption, some
osteocytes are set free from their imprisonment
in the lacunae. These freed osteocytes are
morphologically indistinguishable from resting
osteoblasts, but whether they us actaally relf@rt
back to beiftg osteoblasts is a question that has
not been resolved yet.
Osteoclast
Osteoclasts are large (up to ISO ~m in
diameter), multinucleated cells seen on
surfaces of bone where resorption is taking
place. Usually, they are found in concavities
{Hows)iip's lacunae) that represent areas of
resorbed bone.
In H&E preparations, the cytoplasm
of osteoclasts appears foamy and slightly
basophilic in younger cells, but acidophilic in
older ones. Although there may be two to more
than a hundred nuclei in an osteoclast, only five
to 10 are often seen.
Under the light microscope, the surface
of the osteoclast that abuts on bone is seen to
form a strtared border Eruffledborder). This
ruffled border has been shown by electron
microscopy to consist of villus-like. processes
of plasmalemma, which give off branches that
anastomose with each another. The ruffled
border increases considerably the active
surface of the osteoclast. In addition, it serves
as a contraption where small particles can be
trapped and subjected to enzymatic activity.
Osteoclasts are the cells primarily
responsible for bone resorption (i.e.,
breakdown of bone). Bon.e resorption is
accomplished by the osteoclast at its ruffled
border. Here, the cell pumps H+to acidify the
area and decalcify bone; and releases lysosomal
enzymes that digest collagen and the other
organic components of bone matrix.
For years, the osteoclast had been believed
to arise from the union or fusion of several
monocytes (i.e., white blood cells that serve
as prec.ursor cells for macrophages). B~t
recent scie~tific findings have shown that
multinucleated giant cells that originate from
monocytes are incapable ofresorbing bone. The
emerging view on the genesis of osteoclasts is
that monocytes and osteoclasts have a common
stern cell in the bone marrow, probably
the Colony-Forming Ynit-Granulocy-fe
Macrophage (CEIJ.GM). CFU-GMs that give

Fig. VI-13. Osteoclast (left). The diagram shows an

osteoclast inside a Howships's lacuna (HI). The cell
contains numerous nuclei (nu), mitochondria (mi),
and Iysosomes(Iy). Its surface that abuts on the bone
exhibits a ruffled border (rb).

Fig. VI-14. Osteoclasts (right). The section

shows two osteoclasts which are easy to
identify because they are large and multi-
nucleated. Spongy Bone, H&E x400.

CARTILAGE AND BONE

Fig. VI-1S. Developing Membrane Bone. The section demonstrates bone spicules consisting of
woven bone separated by mesenchyme. The condensed mesenchyme (em) in the lower part of the
photomicrograph will eventually become the periosteum. Note the presence of numerous osteoblasts,
or actively depositing bone matrix, on the surface of the bone spicules. H&E xl 00.

rise to osteoclasts presumably first differentiate
into cellsthat look like monocytes before fusing
together to form osteoclasts.
CARTILAGE FORMATION
(CHONDROGENESIS)
In the embryo, cartilage starts to appear
during the fifth week of intrauterine life. Like
other forms of connective tissue, it arises from
mesenchyme.
At the start of cartilage formation,
the mesencliy:mal cells in the area of
mesenchyme-where cartilage is to develop
(center 0 clIonarificahon)-retract their
cytoplasmic processes, come close together,
multiply rapidly, and then form a mass of
closely-packed cells. In other words, in one
swift continuous process, the mesenchymal
cells differentiate into osteoprogenitor cells
and then, into chondroblasts. As soon as they
are formed, the chondroblasts start to.secrete
the precursors of extracellular fibers and other
organic components of cartilage matrix into
the intercellular area. As they continue to
deposit their secretions all around themselves,
the chondroblasts gradually drift away from
each other and get lodged in lacunae. During
this time, they are referred to as cho drocytes.
.Young clieIl3rocytes still possess mitotic and
secretory capabilities.As chonaroc-ytesmatute,
they become larger and more rounded. At a
certain point they lose their capacity to divide.
Meanwhile, the mesenchy:m that
surrounds the developing cartilage compresses
and differentiates into the perichondrium. The
perichondrium is responsiblefor the subsequent
growth and repair of th~ cartilage..
Cartilage formation starts from the center
of chondrification and proceeds outwards.
Thus, in a developing ,cartilage, mature
chondrocytes occupy the centralarea while
¥-oungchondrocytes and chondroblasts 'occupy
the periphery. Hence, fromthe-center to the
periphery, the cells become progressively
smaller.
Growth of Cartilage
Growth of cartilage is achieved by
two mechanisms: interstitial growth and
appositional growth.

[STEB/\\J & CiO\lZ/\L.ES TEXTBOOK OF HISTOLOGY

Interstitial Growth (Endogenous
Growth)
Intel'-s- iti 1 g owth expands the cartilage
fromwithin. It is onlypossiblein relativelyyoung
cartilage where the intercellular substance is
still malleableand the chondrocytes still possess
the capacity to multiply.
Interstitial growth is heralded by the
mitosis of young chondrocytes. Thereafter, the
daughter cells enlarge the cartilage from within
by secreting precursors of the extracellular
fibers and other organic components of the
matrix into the surrounding intercellular
area.
The secretory capacity of the daughter
cells that arises from the mitosis of a young
chondrocyte is limited. Hence, the amount
of intercellular substance that they are able to
deposit around themselves is likewise limited.
Consequently, the daughter cells, called
isogenous cells, lie close to each other up to
maturity.
" cartilage is invaded by scar tissue (connective
As cartilage ages, its intercellular substance
becomes more rigid. A~ a certain point,
interstitial growth ceases.
Interstitial growth of cartilage is
responsible not merely for the growth of
cartilage from within, but also (as will be
explained later in this chapter) for the growth
in length oflong bones.
Appositional Growth (Exogenous
Growth)
:A':ppositional growt of cartilage is a
function of the 'perichondrium. It occurs when
the osteoprogenitor cells in the perichondrium
multiply and differentiate into chondroblasts.
They then add to the cartilage matrix on the
outer surfaceofthe developingcartilagethrough
their secretions. This manner of growth can be
likened to the way a mason makes a concrete
wall thicker by applying successive layers of
cement on the surface of the wall.
Degeneration and Regeneration
of Cartilage
Cartilage is susceptible to a variety of
degenerative processes. The most- common
of these is rcalcificafion of tlie mafri:xt. It
is a condition that is invariably fatal to the
chondrocytes because when the intercellular
substance calcifies, their nutrition is
compromised.
Calcification of the cartilage matrix is not
always an abnormal phenomenon though; it is,
in fact, one of the initial events that normally
occur in the formation of some bones (see
discussion on endochondral ossification).
With age, cartilage loses its translucency
and becomes opaque. It becomes less cellular
and its matrix less basophilic, indicating a
decrease in its GAG content.
Since cartilage has no blood vessels,
regeneration of the tissue-which is a function
of the perichondrium-is difficult and limited.
In many instances, the damaged area in- a
tissue) instead of being replaced with cartilage.
The regenerative capacity of cartilage is
greater in children than in adults.
BONE FORMATION
(OSTEOGENESIS;
OSSIFICATION)
Bone is produced to replace existing
mesencfi:rme or h~ line cartilage. In the
embryo, the process of replacing mesenchyme
and cartilage with bone starts during the second
month of intrauterine life.
other hand, bone formation that occurs
in areas where a hyaline cartilage exists is
termed endochondnal or intracartilaginous
'Ossification. The bones formed by the first
process are often referred to as memlirane
CARTILAGE AND BONE ..
bones while those formed by the second process matrix likewise traps the osteoblasts in cavities
are called cartilage }iones. These terms are (lacunae),where they become osteocytes whose
misleading because there is actually no physical cytoplasmicprocesses get lodged in tiny tunnels
or chemical difference between membrane and (canaliculi).
cartilage bones. The calcification of bone matrix is a
In either intramembranous or endochondral process that is not fully understood yet.
ossification, bone is first deposited in the form However, the enzyme alkaline I2hosphatase
6f:.woyen }jone. It is a primitive type of bone that osteoblasts secrete may play an important
characterized by high cellularity and random role in its occurrence because soon after
arrangement ofthe collagenfibers in the matrix. it is produced, osteoid starts calcifying.
Woven bone is soon replaced by lamellar berne Incidentally, mature chonarocytes can also
where the intercellular substance is arranged produce alkaline phosphatase; and in the
in thin layers (lamellae) and where relatively presence of this enzyme, cartilage matrix also
few osteocytes are dispersed uniformly. Woy:en calcifies.
Dene is weake , but forms faster than lamellar The area of mesenchyme where the events
pone. described in the preceding paragraphs take
Regardless of location or type of place is referred to as center of ossification.
ossificat-ion,lamellar Doneis initially laid down In the formation of a membrane bone, more
as spongy bone. It is only later that some of the than one center of ossification often appears.
spongy bone is converted into compact bone. In the centers of ossification, as the
osteoblasts on the surface of the bone
-lntramernbranous Ossification spicules continue their secretory activity, the
spicules gradually thicken. The growing bone
fragments eventually fuse together and the
shape of the developing bone emerges.
Meanwhile, the mesenchy:me that
envelops the developing bone differentiates
At the start of intramembranous into the periosteum. At the same time, inside
ossification, the area of mesenchyme where the developing bone, the mesenchyme that
bone will form becomes highly vascularized. immediately surrounds each bone spicule
With the increase in blood supply, the differentiates into endosteum while the tissue
mesenchymal cells differentiate initially into that fills the spaces between the spicules
-osteoprogenitor cells, then into osteoblasts. transforms into my;eloia or Ii_emopoietic
Thus, they become rounded; their cytoplasm tissue.
becomes basophilic (suggesting start of
The periosteum and endosteum play
secretory function); their cytoplasmic
important roles in the subsequent growth,
processes thicken; and the links between
development, remodeling, and repair of bone.
the processes of adjacent cells become
pronounced.
Endochondral Ossification
Soon after they appear, osteoblasts start
laying down components or precursors of Endochondral ossification involves
components ofbona-matrix. Bone matrix, while replacement of a hyaline cartilage model with
still uncalcified, is called osteoi3. Shortly after it bone. It is the type of ossification that forms
has been laid down, osteoid calcifies,giving rise most of the bones of the limbs, pelviS, and
to tiny hone spicules. Calcification of the bone v:erte}jraLc_olllmn.

ESTEB/\)~& GO\jZ,L\L.ES'TEXTBOOK OF HISTOLOGY

 The hyaline cartilage model replaced in
endochondral ossification often resembles the
shape of the future bone.
Endochondral ossification in long bones is
heralded by the appearance of a primary center
of ossification in the middle of the diaphysis.
Changes appear simultaneously in the center of
the cartilage model and in the perichondrium.
At the start of ossification, the
chondrocytes in the center of the cartilage
model hypertrophy and their lacunae
enlarge. They soon begin to secrete alk~line
phosphatase which triggers the calcification
of the cartilage matrix. In as much as the
chondrocytes depend on diffusion of nutrients
from the surrounding intercellular substance
for their sustenance, calcification of the
matrix effectively cuts off their lifeline. Thus,
the chondrocytes die, leaving empty lacunae
(cavities).
Meanwhile, the perich~)lldrium that
envelops the middle area of the diaphysis gets
invaded .by capillaries and becomes highly:
vascularized. In this vascular environment
where the oxygen tension is high, the
perichondriuffit ansforrns into i.' periosteum
and the osteoprogenitor cells differentiate
into osteoblasts. As soon as they emerge,
osteoblasts start laying down components or
precursors of components of bone matrix on
the shaft of the cartilage. The bone matrix
formed soon calcifies. In time, a ring of bone
around the diaphysis, called ~eriosteal bone
\}"--"-ng
(periosteal ~onar), materializes.
Presently, connective tissue, and cellular
elements from the periosteal collar, collectively
referred to as periosteal buil, invade the
cavities left by the chondrocytes in the center
of the shaft. The osteoprogenitor cells in
the periosteal bud attach themselves to the
cartilage remnants. They differentiate into
osteoblasts that start secreting components
or precursors of components of bone matrix.
Shortly thereafter, the bone matrix that has
been laid down calcifies and gives rise to tiny
bone spicules.
In no time, the processes of cartilage
'destruction and bone deposition that have
started in the center of the diaphysis spread
towards both ends of the cartilage. At the same
time, the periosteum thickens and widens the
periosteal bone collar and extends it towards
the epiphyse .
While the ossification process is spreading
towards the epiphyseal ends of the cartilage
model, osteoclasts appear in the central area of
the diaphysis and begin resorbing the bone that
has been previously formed. In the process, the
medullary or marrow cavity is created.
Subsequently, as the bone grows in
circumference through the action of the
osteoblasts in the periosteum, the medullary
cavity also increases in size because of
osteoclastic activity.
Later (i.e., at about the time of birth),
secondary centers of ossificatIon appear
in both epiphyses. Here, the sequence of
events that occurred in the primary center of
ossification is replicated. After a while, only
the articular surfaces of the bone and ~ar~ow
bands of tissue between the diaphysis and
the epiphyses, the e~ipliy-seal pIa es, remain
cartilaginous.
Bone Growth
Unlike cartilage, bone can grow only
byapl!ositional metlio{l because its rigid
intercellular substance does not allow for
interstitial growth or expansion of the tissue
from within. Besides,osteocytes are not capable
of cell division.
Growth in Length of Bone
In spite of the fact that they are incapable
of interstitial growth, long bones can grow in
length, at least until the individual is 20 years old
or so, because of the presence of the epiphyseal
plates.
CARTILAGE AND BONE _
Fig. VI-16. Epihyseal Plate. The cartilaginous
epiphyseal plate separates the epiphysis from the
diaphysis (d). The plate consists of several regions
or zones: resting cartilage cells (r), proliferation
(p), hypertrophy (h), calcification (c)and ossification
(0). Note the periosteum (pe) that envelops the
developing bone. H&E x100.

Epiphyseal Plate
Notwithstanding the appearance of
secondary centers of ossification in both
epiphyses of long bones at about the time of
birth, the bones do not completely ossify until
the individual reaches adulthood. Two thin,
transverse discs ofhyaline cartilage sandwiched
between the diaphysis on the one hand, and the
proximal and distal epiphyses, on the other
hand, persist throughout childhood and the teen
years. Interstitial growth of-these epiphyseal
plates enables long bones to grow in length and
the individual to grow in height.
A longitudinal section through the
epiphyseal plate reveals several successiveareas
(zones) that mirror the stages of endochondral
ossification (i.e., formation of cartilage
followed by destruction and bone formation).
From the epiphysis to the diaphysis, the'seareas
are: 1) zone of resting cartilage cells, 2) zone
of proliferation, 3) zone of maturation or were originally in the zone of proliferation but
hypertrophy,4) zone of calcification, and 5)
were left behind as the cells in the latter zone
zone of ossification.
continued to proliferate and drew away from
The zone of resting cartilage cells anchors them.
the epiphyseal plate to the epiphysis. Its
The zone of calcification is a very thin
moderate-sized chondrocytes are scattered
region. It is only a few cells thick. In this zone,
irregularly throughout the intercellular
the cartilage matrix is calcified and most of the
substance.
cells are already dead.
The zone of proliferation is where
interstitial growth occurs. It is composed of The zone of ossification is the place where
young cartilage cells that continuously undergo bone is deposited through the actions of the
mitosis and expand the cartilage from within. In osteoblasts that have attached themselves to the
this zone, the cells are seen piled on top of each cartilage remnants. In this zone, thin layers of
other like stacks of coins. bone can be seen on the surface of the calcified
cartilage.
The zone of maturation or hypertrophy
consists of large cells and lacunae. The cells While the individual is actively growing,
of this zone do not divide anymore. They the amount of cartilage being produced in

ESTEBAN & CONZ/\LES' TEXTBOOK OF HISTOLOGY

the zone of proliferation is about equal to the
amount being replaced by bone at the zone
of ossification. Hence, the thickness of the
epiphyseal plate is, more or less, kept constant.
When the individual has attained his/her
maximum height, however, the cartilage cells
stop diViding and the cartilage plate is replaced
completely by bone. This event is referred to as
closure of the epiphyseal plate (closure of the
epiphysis). It signifies the end of the increase
in length of the bone. When all the epiphyses in
all the long bones of the body have closed, the
individual stops growing in height.
Growth in Width of Bone
Growth in width ofabone is afunction ofthe
periosteum. It occurs when the osteoprogenitor
cells in the osteogenic layer differentiate into
osteoblasts, which then add layer upon layer
of bone matrix on the outer surface of the
bone.
In growing individuals, cortical bone is
continually added to the surface of a long bone
to ensure that every increase in bone length
is associated with a proportionate increase in
width or diameter. However, during this time,
the thickness of the bone does"not increase
appreciably because the medullary cavity is
continuously enlarged through resorption by
the osteoclasts.
Conversion of Spongy Bone
into Compact Bone
Lamellar bone, as previously mentioned, is
first laid down in the form of spongy bone.
In spongy bone, the bone spicules are
connected together to constitute the framework
of a very complicated network of spaces in the
form of irregular tunnels. To form compact
bone, successive layers of bone matrix are
deposited on the inner surface of the tunnels
by the osteoblasts in the endosteum, until
only a small canal (Haversian canal) occupied
by blood vessels remains of the original
tunnel.
Bone Remodeling
Boneis avery dynamic tissue that constantly
undergoes two forms of remodeling: external
and internal.
External Remodeling
External remodeling refers to changes that
occur in the shape of a bone in response to
external factors that affect it. It is accomplished
by' bone resorption in certain areas and
deposition of bone tissue in other areas. An
exampleof external remodeling is the formation
of tubercles in some long bones in response to
the pull of muscles.
Internal Remodeling
Internal remodeling refers to the never-
ending cycle of resorption and formation of
Haversian systems. It is necessary because of a
couple of reasons. '
Firstly, the supply route for nutrients and
oxygen to the osteocytes, even in the best-
formed Haversian systems, is very inefficient.
Hence, at any given time in some Haversian
systems,many ofthe osteocytes are already dead
or dying, and the remaining ones are incapable
ofmaintaining the intercellular substance ofthe
system. Thus, these Haversian systems have to
be renewed or remodeled. Renewal ofHaversian
systems is a two-stage process: 1) resorption of
existing systems followed by 2) formation of
new ones in the resorbed areas. Incidentally, as
previouslymentioned in this chapter, interstitial
lamellae are probably partially resorbed
Haversian systems.
Secondly,when there is a drop in the blood
calcium level, some of the Haversian systems
have to be resorbed to mobilize the calcium
that they contain. These sacrificed Haversian
systems are rebuilt during "times ofplenty."
Nutritional Effects on Bone
Dietary deficiency of either calcium or
phosphorus leads to poor mineralization
CARTILAGE AND BONE
 of bones. The bones consequently become
brittle and fracture easily.
Vitamin D deficiency results in poor
intestinal absorption of calcium. When this
happens; newly formed bone matrix (osteoid)
fails to calcify, resulting in softened bones. In It inhibits bone resorption
children, this failure of calcification results in . suppressing osteoclasts. Calcitonin is secreted
a condition called rickets. It is characterized
by stunting of growth and developing weak
bones. In adults, it 'gives rise to osteomalacia
characterized by lack ofbone mass and frequent
fractures. i
Vitamin C deficiency causes scurvy, a
disease condition that is characterized by poor
production of bone matrix.
Vitamin A deficiency adversely affects the
osteoblasts and osteoclasts and retards bone
growth.
Endocrine Effects on Bone
One of the functions of bone is to serve
as a storehouse for calcium, one of the most
important minerals in the body. Calcium
is essential for blood coagulation, muscle
. contraction, nerve function, and manyother
vital metabolic processes. It is, therefore,
imperative for the blood calcium level to be
maintained within a certain range.
The primary regulator ofblood calciumlevel
is the parathyroid hormone (parathormone)
produced by the parathyroid gland. A drop in
blood calcium level stimulates the parathyroid
gland to secrete parathormone whose target
cells are the osteoblasts. Parathormone has a
dual effect on osteoblasts: it inhibits their bone-
forming activity and stimulates them to secrete
the hormone osteoclast-stimulating factor.
ESTE3;\N & CO",IZf\LES' TEXTBOOK OF HISTOLOGY
As its name implies, the latter stimulates the
osteoclasts to accelerate their bone resorbing
activity.
Opposing the activity ofthe parathormone
is the hormone calcitonin (thyrocalcitonin).
by directly
by the parafollicular cells of the thyroid
gland and perhaps some other still unidentified
cells of the body.
The gonadal hormones play an important
role in determining the rate of skeletal
maturation. Precocious sexual development
results in the premature closure of the
epiphyseal plates and stunted growth. In
females, estrogen increases the number
of osteoblasts and thus, stimulates bone
formation. In actively growing males, the
absence of' a d ogens (as in castrated
individuals or eunuchs, for example) delays
the closure of the epiphyseal plates, resulting
in disproportionately long limbs.
Humallgrowtliliurmone thGH] secreted
bythe pituitary gland also markedly influences
bone growth. In growing individuals, the
absence or low levels of circulating growth
hormone results in cessation of mitoses of
the chondrocytes at the epiphyseal plate and
consequently, dwarfism. Meanwhile, an
excess of the hormone results in enhanced
mitosis of the chondrocytes in' the epiphyseal
plates and, therefore, in gigantism. In adults,
in as much as the epiphyseal plates have already
closed, excess of growth hormone does not
result in increase in height. Instead, it leads
to acromegaly, a condition characterized by
deformed bones including overgrowth and
protrusion of the mandible (lower jaw).
Blood
lood, which is generally considered by
B histologists as a special type of loose
\connectLve tissut!, is the fluid that
circulates in the cardiovascular system (i.e.,
heart and blood vessels). Grossly, when
oxygenated, as that in systemic arteries, blood
is light red, but when unoxygenated, as that in
systemic veins, it is dark red to purple.
Like all connective tissues, blood consists
of cellular elements and intercellular substance'.
The intercellular substance of blood is a fluid
called plasma while the cellular elements, which
are collectively referred to as formed elements,
include red blood cells, white blood cells, and
platelets.
Blood transports oxygen from the lungs
and nutrients from the gastrointestinc,ll tract to
tissues where they are needed. It also carries
carbon dioxide and waste materials to the
organs where they are to be disposed of. In
addition, it carries other substances (hormones,
proteins, etc.) to their target tissues or organs.
Furthermore, it plays a major role in defending
the body against pathogenic microorganisms
and toxic substances by providing the cells and
transporting the substances needed for the
purpose.
In the field of medicine, blood is the
tissue that is most widely used for diagnostic
and monitoring purposes This is so because
the various components of blood give direct
and indirect clues to the state of health of the
different organs and tissues. Furthermore,
obtaining a blood sample is fairly easy and.
relatively harmless.
Blood comprises about 8% of a person's
body weight, which roughly translates to 5.0
liters for a 60 kg person. It is slightly alkaline
with a normal pH of7.4.
When blood is drawn and placed in a test
tube that contains an anticoagulant like heparin
and then centrifuged, its constituents separate
and form three layers. The top layer, which
usually comprises about 54% of the volume of
the column, is plasma: The middle layer, called
buffy coat, is a thin white film that is formed by
the platelets and white blood cells. It comprises
only about 1% of the column. The bottom layer
is red in color because it contains the red blood
cells. It accounts for about 45% of the column.
Thus, the formed elements account for about
46% of total blood volume.
On the other hand, when blood is drawn
and placed in a test tube that does not contain
an anticoagulant and left to stand, it forms
a clot-a jelly-like mass consisting of fibrin,
a protein that forms a fibrous network, some
blood clotting factors, and the formed elements
that get entrapped in the fibrin. Incidentally,
fibrin is produced by the action of thrombin on
 fibrinogen, one of the plasma proteins. If the
clot is removed from clotted blood, the straw-
colored fluid that remains is called serum.
Serum, therefore, is plasma minus fibrinogen,
the formed elements, and some blood clotting
factors that participate in the formation of the
clot.
Lipoproteins are plasma proteins that
transport lipids, which are insoluble in water,
from the intestines to the liver and from the
liver to the tissues while the complement
system consists of more than 20 proteins that
are involved in the inflammatory and immune
responses (processes).

PLASMA FORMED ELEMENTS OF

BLOOD
Plasma is a transparent, yellowishfluid that
contains numerous dissolvedsubstances. Water The formed elements of blood (Fig. VII-
comprises 90% ofits volume while the dissolved 2 to Fig. VII-10) consist of cell fragments and
substances-organic compounds (chiefly cells. The cell fragments consist of platelets
proteins), inorganic salts and ions-account (thromboplastids) whilethe cellsare composed
for the other 10%. of red blood cells (RBCsj erythrocytes:
rubricytes) and white blood cells (WBCsj
There are over a hundredplasma proteins
leukocytes) .
including albumin, globulins, fibrinogen,
lipoproteins, the complement system, and a WBCs are classified into two groups
variety of regulatory proteins such as enzymes, (granular and agranular) based on the presence
proenzymes, and hormones. Albumin or absence of specific cytoplasmic granules
accounts for 60%,globulins 36%,fibrinogen 4% in their cytoplasm in routinely-stained blood
and the rest less than 1%,of all plasma proteins. smears. Granular leukocytes (granulocytes)
All the plasma proteins are synthesized by the c~~tain specific granules in their cytoplasm
while agranular leukocytes (agranulocytes)
liver except for the gamma globulins which are
do not.
. produced by plasma cells and the regulatory
proteins which are synthesized in various Incidentally, the Romanowsky method
organs. is the prototypical staining technique that
is used to distinguish the formed elements of
Albumin, which has a molecular weight of
50,000, is the smallest of the plasma proteins. It
maintains the colloidosmotic pressure ofblood.
It also binds and transports some molecules,
notably free fatty acids, some steroid hormones
and bilirubin.
The globulins consist ofthree kinds: alpha,
beta, an? gamma. With molecular weights that
range from 80,000 to 1million, the globulins are
larger than albumin. Alpha and beta globulins
bind and transport substances to various
parts of the body while gamma globulins
(immunoglobulins) comprise the antibodies
of the immune system.
Fibrinogen is the precursor of fibrin, the
protein needed to complete the final step of Fig. VII-1. Normal Volume of the Different
Componentsof Blood
blood clotting.

ESTE8i\N& (jOND\LES' TEXTBOOK OF HISTOLOGY

 Number and Volume of the
Formed Elements of Blood
The overwhelming majority of the formed
elements of blood are red blood cells (RBCs).
Normally, there are 4 to 6 million RBCs per
cubic millimeter (cu mm) of blood. Males have
4.5 to 6.0 million/cu mm while femaleshave 4.0
to 5.5 million/cu mm.

Tabfe VII-1. Normal Number of Formed

Elements in Blood

Fig. VII-2. Platelets (at arrow) and RBCs

(surrounding cells). Blood Smeer, Wright x1,OOO.
Incidentally,the percentage ofblood volume
blood and cells in the bane marrow from each that is accounted for by the RBCs (which, as
other under the light microscope with the use already mentioned, is usually about 45%) is
of acid and basic dyes. It is the forerunner of referred to as hematocrit, a very important
several similar methods that are currently used measure in clinical medicine. Hematocrit
for routine staining of blood and bone marrow (HCT; hct; Ht) is an indirect gauge of the
smears including Giemsa, Jenner, Wright, and· number of RBCs present in blood. It is one of
Leishman methods. the parameters used to screen for and measure
the extent of anemia-a condition where the
The granulocytes are further classified
oxygen-carrying capacity ofblood is decreased.
into three kinds-neutrophil, eosinophil, and
basophil -on the basis of the affinity for dyes
of their specific cytoplasmic granules.
The agranulocytes, on the other hand,
consists of two kinds: monocyte and
lymphocyte.
There are three types of lymphocytes:
T lymphocyte (T-cell), B lymphocyte
(B-cell) and natural killer cell (NK cell)
but they look alike under the microscope.
They are distinguishable only with the use of
special histochemical techniques including
immunocytology.
Of the formed elements of blood, only
platelets and RBCs carry out their functions
within the cardiovascular system. The WBCs
are only in transit in blood; they invariablyleave
the bloodstream to enter connective and other Fig. VII-3. Neutrophil. Indicated by the arrow is a
tissues where they carry out their functions. Barr body. Blood Smear, Wright x1,OOO.

BLOOD 'I4iI
__ - - __ - ---------------------------------------------------------~
A person's hematocrit is considered normal if it
falls within a certain range. This range depends
on age and sex, and varies slightly among
laboratories. In adults, the normal hematocrit
range is 40% to 54% in males and 37% to 47%
in females.
In normal blood, the number of platelets is
between 150,000 to 400,000/ cu mm of blood.
The WBCs are the fewest among the
formed elements. Their normal number is only
4,500 to lO,500/cu mm of blood.
Fig.VII-4. Neutrophils (ne),Lymphocyte(Iy),and
Platelet (pi). Blood Smear,Wright x1,OOO.
WBC Differential Count
WBC differential count refers to the
percentage distribution in blood of the five
types ofWBCs. The average normal values for
this parameter are: neutrophils =50%-70%,
lymphocytes = 20%-40%, monocytes = 3%-7%,
eosinophils = 2%-5% and basophils = 0%-1%.
Table VII-2. WBC Differential Count: Average
Normal Values
Neutrophils 50-70
Eosinophils 2-5
Basophils 0-1
Lymphocytes 20-40
Monocytes 3-7
The WBC count and differential count are
very useful clinical indices because they provide
ESTES/\f'-..!& CjO~~Z!\LES'TEXTBOOK OF HISTOLOGY
clues into the diagnosis and help in monitoring
the course of many diseases. For example, in
acute bacterial infection, there is leukocytosis
(i.e., the WBC count is elevated) and the
WBC differential count shows neutrophilia
(i.e., increased percentage of neutrophils). In
contrast, in certain viral infections, there is
leukopenia (i.e., the WBC count is less than
normal).
Shapes of the Formed Elements
of Blood
The red blood cells are biconcave
structures, but they appear round when viewed
on their flat surface. Platelets, on the other hand,
are biconvex discs when seen in coronal view but
ovate when seen in transverse view.
The white blood cells are spherical when
they are in circulating blood, but in the tissues,
they assume various forms and are amoeboid.
Sizesof the Formed Elements
of Blood
The red blood cells are small cells that have
an average diameter of7.5 ~m and a thickness of
1.9 ~m. Since they are present in practically all
histologic preparations, they are routinely used
as a built-in yardstick for estimating the sizes of
structures in any given microscopic field.
Table VII-3. Normal Sizes of the Formed
Elementsof Blood
 Platelets are the smallest of the formed
elements of blood. They are only between 2-3
~m in size.
WBCs are spherical cells that are larger in
blood smear preparations than in vivo because
they flatten out when blood is spread on glass
slides.
N eutrophils and basophils are of the same
size. Their average diameter in circulating blood
is 7.0~m, but in blood smears, they are between
10 to 12 ~m.
Eosinophils are merely 9.0 ~m in diameter
in circulating blood but 12to 14~m in smears.
The monocytes are only between 9-12 ~m
in diameter in circulating blood, but in dried
smears, at 17-20 ~m, they dwarf the other
formed elements.
Lymphocytes consist of two kinds in terms
of size: small and large. Most lymphocytes are
small lymphocytes that are 6-9 ~m in diameter
in circulating blood but 8-11 ~m in blood
smears. The large lymphocytes are 9-12 ~m in.
diameter in circulating blood and 11-15 ~m in
blood smears.
Lifespan of the Formed Elements
of Blood
The average lifespan of RBCs is 120 days
while that of platelets is just between 9-12 days.
N eutrophils stay an average of 8 hours in
blood. Then they leave the blood and reside and
work for another 1-4 days in connective tissues.
Eosinophils, on the other hand, circulate in the
blood for 3-8 hours and then migrate to other
tissues where they survive for 8 to 12 days.
Basophils, meanwhile, live only for a few days.
Monocytes stay in circulating blood for 1
to 2 days, then escape from the blood vessels
and migrate to connective tissue where they
differentiate into macrophages and live for
another 70 days ( 6-16 days according to some
authors.)
Table VII-4. Normal Lifespans of the Formed
Elementsof Blood
Lymphocyte
B
T
NK
Monocyte 1-2 days
The amount of time lymphocytes spend
in blood is unknown because unlike the other
WBCs, which stay permanently in the tissues
after they leave the bloodstream, lymphocytes
"re-circulate." But it has been established that
lymphocytes are long-lived; B-cells live for
several months; and T-cells survive for several
years.
Red Blood Cell (RBC;
Erythrocyte; Rubricyte)
The red color of blood is due to the RBCs
which, when seen as individual cells, are actually
not red. The unstained RBC is pale yellow or
greenish yellow while the routinely-stained
one is pink. The color of RBCs is imparted
by hemoglobin (Hb; hb; Hgb), the oxygen-
carrying pigment of the cells.
Mature RBCs are anucleate and devoid
of cytoplasmic organelles. They are simply
membrane-bound corpuscles whose cytoplasm
is loaded with dissolved hemoglobin, the
component of the cells that is responsible
for the main function of the RBCs, which is
the transport of oxygen from the lungs to the
different tissues of the body.
RBCs are produced in the bone marrow.
Before entering the blood, they extrude their
nucleus and all their cytoplasmic organelles.
However, not all RBCs in the blood are
mature cells. It is normal to find about 0.8% of
Circulating RBCs in the form of immature cells
BLOOD 'IiiiI
called reticulocytes (polychromatophilic
erythrocytes). Reticulocytes are slightly
bigger than mature RBCs. They do not have a
nucleus but they still contain a few cytoplasmic
organelles including ribosomes which impart
a bluish tinge to their cytoplasm when routine
staining is applied.
Hemoglobin is a globular protein that
normally comprises about 33% of the mass of
RBCs. It consists of a protein molecule (globin)
and an iron-containing compound (heme). In
the lungs, oxygen combines loosely with heme
after which it is transported to and released
in the tissues. By the way, although some
amount of carbon dioxide reacts directly with
hemoglobin, blood transports carbon dioxide
from the tissues to the lungs mainly in the form
of bicarbonates that are dissolved in the plasma.
The amount of hemoglobin (Hb) perIOO ml
ofblood is used in clinical medicine as a measure
of the oxygen-carrying capacity of blood. The
normal amount of hemoglobin in males is 14-18
g per 100 mL of blood while in females, it is 12-
16 g per 100 mL.
In routinely-prepared blood smears, the
depth of staining of RBCs reflects the ,amount
of hemoglobin they possess. RBCs that contain
the normal amount of hemoglobin are pinkish
in color and are termed normochromic. Those
that contain less than the normal amount of
Fig. VII-5. Neutrophilic Band Form (at arrow).
Blood Smear; Wright x 1,000.
ESTEB/\P-,J& (iONZALES' TEXTBOOK OF HISTOLOGY
hemoglobin are paler and are referred to as
hypochromic while those that have more than
the normal amount of hemoglobin stain more
intensely and are termed hyperchromic.
In some disease conditions, RBCs manifest
variations in size (anisocytosis). Normal-
sized RBCs are called normocytes. RBCs that
are greater than 9 ~m in diameter are called
macrocytes while those that are less than 6 ~m
are called microcytes.
RBCs are very pliable and may momentarily
fold to be able to pass through small vessels.
In vitro, they tend to adhere to each other on
their flat surfaces like a stack of coins. This
phenomenon, known as rouleaux formation,
is a very co-mmon artifactual finding in thick
smears, but when noted in a thin smear (true
rouleaux), it could signify an increased amount
of plasma proteins, particularly fibrinogen and
globulins, on the surface of the RBCs that make
the cells stick together. True rouleaux formation
occurs in multiple myeloma, malignant
lymphoma, chronic infection, and a few other
disease conditions.
When placed in a hypotonic solution, RBCs
undergo hemolysis. In this process, the cells
initially swell and become spherical because of
the entry of water. Their cell membrane then
stretches and allows hemoglobin to leak out.
This reduces the cells into pale membrane-
bound structures called '(e~throc__-y:te
ghosts."
Sometimes, RBCs form surface spicules or
spines and are called echinocytes. This process,
called crenation, was once thought to be due
to hypertonicity of the environment. It is now
attributed to low ATP levels.
RBCs are normally biconcave discs. In
certain disease conditions, however, they
exhibit variation in shape ~poikdocy-tosis~-
they could be ellipsoidal, spherical, sickle-
shaped, oval, teardrop-shaped, etc.
The cell membrane of RBCs contains a
variety of antigens (agglutinogens) that form
the basis for the classification of blood into
30 blood group systems by the International
Society of Blood Transfusion. An antigen is a
substance which the immune system perceives
as foreign to the body and which consequently
induces an immune response that, in the case of
RBC antigens, comes in the form of production
of antibodies (agglutinins) that, with the
help of other factors, destroy the antigen. In
clinical medicine, the most important antigens
of RBCs are those that comprise the ABO and
Rhsystems.
In the ytu:o s¥stem (~BO 6100d
group-ing~, there are two kinds of antigens: ~
and B. In this system, blood type is determined
on the basis of the presence or absence of one
or both antigens. Persons whose RBCs contain
antigen A have ~pe ts. btOOG; those with B
Fig. VII-6. Lymphocytes (Iy), Neutrophil (ne), and
antigen have t}':pe:'Bblood; those that possess
Platelets (pi). Blood Smear, Wright x1,OOO.
both antigens have tY12e:ABblood; and, those
that have neither antigen have t}':pe e blood.
Incidentally, in theAb O system, a person's mitochondria, ribosomes, elements of Golgi
plasma contains naturally occurring antibodies complex, and rough and smooth endoplasmic .
against the antigens that are not present in reticulum.
the person's RBCs. Thus, t}':pe persons have In routine LM preparations, a platelet
anti -B antibodies in their plasma; ty-~ people is seen to consist of a dark-staining central
have anti-A antibodies, t},:12eG. people have portion (granulo-mereJ that contains
both anti-A and anti-B antibodies while t}':pe basophilic granules that are often referred to
7\:Bpersons have no antibodies. as azuro fiilic granu es and a clear peripheral
portion Ehxalomer }.
In the Rh s~stem (RH blood g.l"ouping~,
people with the Rh antigen on the surface of Electron microscopy and special histologic
their RBCs are designated Rfi positiv:e 'RB ) techniques have shown that the granules that
while those that do not have the Rh antigen are dot the granulomere are secretory vesicles.
classified Rh negaJiv:e C:lth-~.Unlike the ABO These vesicles contain hydrolytic enzymes that
system, the Rh system is not characterized by are similar to those in lysosomes and a variety
naturally-occurring antibodies. Hence, Rh of other compounds including platelet factor 4
individuals do not have naturally-occurring (PF-4), platelet-derived growth factor (PDGF),
anti-Rh antibodies in their plasma. thrombospondin, and von Willebrand factor.
The granulomere also contains mitochondria,
albeit few.
Blood Platelets
(Thromboplastids) Electron microscopy has also revealed that
the platelet membrane forms numerous tubular
Platelets (Fig. VII-2) are membrane- invaginations ts-urfa_ceconnecting canalicular
bound cell fragments that are often seen in rs-y:stem.)This canalicular system connects the
clumps in blood smears. They are produced by interior to the surface ofthe platelet and serves as
fragmentation of the cytoplasm of giant cells entry and exit points of substances to and from
known as megal{aq_::ocytes. They have no the platelet. Interspersed among the elements
nucleus but they contain cytoplasmic organelles: of the canalicular system is a set of narrower

BLOOD·.
 channels termed dense tubular system. The
dense tubular system is the main site for storing
Ca ions and cyclooxygenase, the enzyme that
converts arachidonic acid to precursors of
prostaglandins and thromboxanes, substances
that mediate a variety of physiological
processes including the inflammatory response
(inflammation).
A platelet has a cytoskeleton consisting of
microtubules and microfilaments that serves as
a framework to anchor the platelet membrane
and allow signal transduction to take place.
Platelets play an important role in
hemostasis) i.e., the arrest of bleeding after
injury to a blood vessel. When a blood vessel is
injured) platelets adhere to the exposed collagen
fibers) a process called elatelet a esion. After
adhering to collagen fibers) platelets release a
substance that promotes J2latelet aggregatjpn)
causing platelets to stick in increasing numbers
to those that have already adhered to the injured
vessel. Aside from participating in platelet
adhesion and aggregation) platelets also release
the blood clotting factors that are contained in
their secretory granules.
Neutrophil (Neutrophilic
leukocyte)
The most distinctive feature of neutrophils
(Figs. VII -3 and 4) in LM preparations is the
presence in their cytoplasm of fine specific
granules that have little affinity for dyes.. The
specific granules are rather small (0.1 ~m in
diameter) to be appreciated individually under
the light microscope. However) they impart a
grainy texture and a lilac tinge to the cytoplasm.
Incidentally) specific cytoplasmic
granules are so-called because they are specific
to (i.e., found only in) granulocytes. They are
actually secretory vesicles.
The specific granules of neutrophils
contain some enzymes and proteins that have
bactericidal properties (collectivelyreferred to as
phagocytin_s) and compounds that mediate the
inflammatory response. When released) these
ESTEB!\N& CCNZf\L.ES'TEXTBOOK OF HISTOLOGY
~ --- ----------------------------------------------------------------
attract eosinophils, monocytes, neutrophils,
and other cells involved in the inflammatory
process) and increase the permeability of blood
vessels.
In addition to specific granules) neutrophils
also possess nonspecific c:y:to]llagnicg-ran-ules
~a_zurophilicgranule ). Azurophilic granule is
the generic term for any cytoplasmic granule
that stains with azure or a similar blue aniline
dye. Azurophilic granules are present in a
number ~f cell types) including all WBCs) and
their content varies depending on the cell type.
In WBCs) the azurophilic granules) which stain
reddish-purple in routinely-stained smears) are
lysosomes.
Among granulocytes) therefore) the specific
granules contain substances that are destined
to be exported by the cell while the azurophilic
granules contain lysosomal enzymes that are
intended for use within the cell.
The azurophilic granules of granulocytes
are sometimes referred to as ~rimary; granules
because during the differentiation of these cells)
they appear earlier than the specific granules
Esecunaarygranules).
In neutrophils, the azurophilic granules)
which are about O.S ~m in diameter) are larger
but fewer than the specific granules.
The nucleus of neutrophils forms 2-6 lobes
that are joined to each other by a narrow band
of nuclear material. It is deeply staining and
usually has no visible nucleolus. The number
oflobes of the nucleus increases with the age of
the cell. Young neutrophils have only two lobes
in their nucleus while old ones have up to six.
In females) at least 6 out of sao neutrophils
contain a nuclear appendage known as
drumstiok or.Barr b.o.aythat is attached to one
of the lobes of the nucleus. The" drumstick"
is a small nuclear fragment that represents
the condensed) inactive X chromosome. In
males) the drumstick may also be present but
never up to 6 out of sao neutrophils.
It is not unusual for some immature
neutrophils (tj_ana tOFmSj stab cel ls)
(Fig. VII-S) to be able to enter the bloodstream
even before their nucleus has formed lobes.
The nucleus of these cells is usually elongated
and horseshoe-shaped or S-shaped. Stab cells
c<?nstitute 2%-8% of neutrophils in normal
blood.
N eutrophils defend the body against
pathogenic bacteria and other foreign substances
that have penetrated the body's surface
barriers (i.e., skin and mucous membranes) by
phagocytosis (with the use oflysosomes) and
by releasing the substances in their specific
granules that are toxic to pathogens. Neutrophils
also mediate the inflammatory response
because some of their secretory granules contain • Fig. VII-7. Eosinophil (at arrow). Blood Smear,
mediators' of the" inflammatory" response." . Wright x1,OOO.
They also serve as APCs either indirectly
(i.e., apoptotic neutrophils containing foreign smears. These coarse, refractile, uniformly-
protein can be phagocytosed by dendritic cells)" . sized, and intensely eosinophilic granules fill the
or directly by presenting antigens to T-cells """cell,making it difficult to appreciate the nucleus
themselves. But, as APCs, they are not as. at times. The nucleus of eosinophils contains
efficient as macrophages. only 2 to 3 (usually 2) lobes. Its chromatin
material is finer than that of neutrophils.
N eutrophils are highly motile cells that
move about the extracellular spaces with the- The specific cytoplasmic granules of
aid of pseudopodia. They are attracted by eosinophils contain several distinct cationic
chemicals (chemotaxins) that are generated by proteins and a variety of hydrolytic enzymes.
bacteria and other cells as a reaction to bacterial The azurophilic cytoplasmic granules of
components, or released by certain cells in eosinophils are fewer and smaller than their
damaged tissue. specific granules.
Mature neutrophils are terminally Eosinophils have limited phagocytic
differentiated cells that have very few activity. They do not phagocytose bacteria.
cytoplasmic organelles. Consequently, they are Instead, they defend the body against
not able to regenerate their expended enzymes microorganisms by phagocytosing and breaking
and other proteins. As a rule, after the contents down antigen-antibody complexes.
of their granules are spent, they die and form
Eosinophils also defend the body against
part of pus-if pus formation occurs-where
parasites especially roundworms by releasing
they are known as pus cells.
the content of their granules in the extracellular
environment. Some of their granules contain
Eosinophil (Eosinophilic
substances that are directly toxic to certain
Leukocyte) parasites and their ova. Similarly, they destroy
Eosinophils (Fig. VII -7) are only slightly cancer cells.
larger than neutrophils, but under the LM, they However, although eosinophils function
can easily be distinguished from the latter by to protect the body, some substances in
the presence of large specific granules in their their granules, when released, amplify and
cytoplasm that stain pink to brick red in routine perpetuate the body's inflammatory response

BLOOD
 (inflammation) leading to worsening of
symptoms in such conditions as asthma and
allergy.
The eosinophils in blood dramatically
increase in number in instances of heavy
parasitic infestation and allergy.
Basophil (Basophilic Leukocyte)
Basophils (Fig.VII-8) are about as large as
neutrophils. They can easily be distinguished
from the latter in routine LM preparations
because their specific granules are coarser
and stain blue or dark purple. The specific
cytoplasmic granules of basophils vary in
size, but they are larger, albeit fewer, than
Fig. VII-S. Basophil (at arrow). Blood Smear,
Wright x1,OOO.
those present in eosinophils. They are water-
soluble and some are washed away by aqueous
stains. They contain histamine, heparin, and
leukotrienes.
The nucleus of basophils is usually
U-shaped or J -shaped, but sometimes it appears
bi-lobed. It is often difficult to appreciate in
routinely-stained blood smears because it is
hidden by the cytoplasmic granules.
Basophils are similar to mast cells of
connective tissue in that both cell types have
cytoplasmic granules that contain histamine
and heparin. The two cell types, however,
[SlTU/\f,j & CiO\Z/\L.ES' TEXTBOOK OF HISTOLOGY
-- -- --------'---- ------------
arise from different progenitor cells and differ
markedly in many aspects. Compared to mast
cells, basophils are smaller but more mobile;
their cytoplasmic granules are larger but fewer;
and, they have a shorter lifespan.Also,mast cells
can divide while basophils cannot.
In terms of function, the basophils are
the least known among the granulocytes.
But it is known that when activated, they
release histamine and the other mediators
of inflammation that are in their granules.
Like mast cells, they are also involved in the
pathogenesis or development of immediate-
type hypersensitivity reactions including
anaphylactic shock.
Monocyte
In routine LM preparations, monocytes
have a bean-shaped or U-shaped nucleus that
is usually eccentrically located. Often, two or
more nucleoli can be seen in the nucleus.
Monocytes have abundant basophilic
cytoplasm that contains numerous azurophilic
granules.
The function of monocytes is to serve as
precursor cells for macrophages'.
Monocytes originatefromthe bone marrow,
stay in blood for a while, and then migrate into
connective tissue where they differentiate into
rnacrophages.
Fig. VII-9. Monocyte (nucleated cell) and Platelets
(at arrow). Blood Smear, Wright x1,OOO.

Fig. VII-10. Large Lymphocyte (at arrow). Blood
Smear; Wright x1,OOO.
lymphocyte
Lymphocytes comprise a family of cells that
act as the principal agents of the body's immune
response.
At any given' time, about 98% of
lymphocytes in the body are not in blood but
in other tissues and different lymphoid organs.
The lymphocytes that are in blood, to reiterate,
can be classified into two types according to.
size:" mal and 1 _rge,but the vast majority of
lymphocytes (97%) are small lymphocytes.
,.
In routinely prepared blood smears, a
small lymphocyte (Fig. VII-4 and 6) is seen as
having a high nucleus to cytoplasm ratio. Its
nucleus occupies almost the entire cell. The
nucleus is kidney-shaped or round and contains
coarse, clumpy chromatin granules that are
intensely staining. The cytoplasm of the small
lymphocyte consists of a thin, basophilic "halo"
that surrounds the nucleus. It has no specific
granules but it contains a few azurophilic
granules.
Electron micrographs of small lymphocytes
have revealed that their most conspicuous
cytoplasmic organelles are the numerous free
ribosomes that are responsible for the basophilia
of the cytoplasm. The other organelles-a
small Golgi complex, a very poorly developed
endoplasmic reticulum, a pair of centrioles and
one or two mitochondria-are concentrated in
the region of the concavity of the nucleus.
In contrast to small lymphocytes, large
lymphocytes (Fig. VII-lO), in routine blood
smears, are noted to contain abundant
cytoplasm that is intensely basophilic. Their
nucleus is typically large but relatively pale. It
contains a very prominent nucleolus.
In electron micrographs, large lymphocytes
are .se en to have a well-developed Golgi
complex, rER, and numerous mitochondria and
lysosomes.
Large lymphocy.tes are often mistaken
for monocytes in routine LM preparations.
However, compared to large lymphocytes,
monocytes are generally bigger. In addition,
monocytes have more abundant, although
lighter-staining, cytoplasm. Their nucleus
is also lighter staining than that of large
lymphocytes because its chromatin material is
less condensed.
. The large lymphocytes are probably not
young cells, as once thought. The current
prevailing opinion is that they are either NK
cells, which are inherently larger than small
lymphocytes, or activated lymphocytes (Note:
Lymphocytes enlarge when activated) that are
on their way to the lymphoid tissues and organs
where they will proliferate and differentiate into
plasma cells and memor:y:B cells.
The amount of cytoplasm of a lymphocyte
is directly related to its state of activity. Thus,
small lymphocytes, as a rule, are inactive.
Classification of Lymphocytes
Lymphocytes look alike under the
microscopes, but on the basis of antigen
receptors that are present on their surfaces,
which can be detected byusing special histologic
techniques including immunocytology, they are
classified into three general types: B-cell, T-cell
andNKcell.
BLOOD ,..
 T-cells carry T-cell antigen receptors
(TCR) while B-cells carry B-cell antigen
receptors (BCR). NK cells were for a while
believed to carry no surface antigens but it
is now, known that they also express unique
surface receptors called natunl c~y:totoxicit:y
receptors ~NeRsy. Furthermore, NK cells also
carry certain receptors that are present in some
T-cells.
.. Of the lymphocytes in blood, 80% are
T-cells,15%are B-cells,and the rest (5%)are NK
cells. The two main types oflymphocytes (i.e.,
B-cell and T-cell) differ in their developmental
background, function, and lifespan. The
developmental background and lifespan of
natural killer cells (NK cells) have not been
established yet. . perforin and proteases called granzymes into
B-cells are responsible for humon}
immunity, one of the two types of immune
response the body exhibits (see chapter on the
lymphoid system).
T-cells, on the other hand, are responsible
for c-e:lI-meaiatea immunif'y GceJJular
immunity)-the other type of immune
response that the body manifests. In addition,
some T-cells also participate in effecting a
humoral response or immunity.
ESTEBi\\j & CJOf\iZN ..ES' TEXTBOOK OF HISTOLOGY
T-cells are classified into several types
based on their functions and surface markers:
1)helper T-cell or T h -cell, the most numerous
among the T-cells; 2) cytotoxic T-cell (Tc-
cell); 3) memory T-cell; and, 4) suppressor
T-cell (Ts-cell). The functions ofthese different
types of T-cells are discusse~ in the chapter on
lymphoid tissue.
NK cells are cytotoxic cells that play an
important role in the body's inflammatory and
immune responses. They destroy tumor cells
and cells infected by viruses.
NK cells do not produce antibodies. They
kill their target cells by releasing the content
of their cytoplasmic granules consisting of
the intercellular space. The proteins that they
releasekill target cellsin the vicinity by inducing
apoptosis (i.e., programmed cell death) or
osmotic cell lysis.
NK cells are effective in containing viral
infections while the B- and T-cells are still
mounting a decisive immune response.
NK cells also serve as antigen presenting
cells (APCs)-which are discussed further in
the chapter on the lymphoid system.
Hemopoiesis

he formed elements of blood have of the bone marrow, except for lymphopoiesis

T relatively short lifespans. On any given -whichoccurs not only in the bone marrow but
day, numerous blood cells die and the in all1r-m-phoidtissues and organs.
body has to continuously produce new ones to
maintain their numbers. It is estimated that in HEMOPOIETIC TISSUE
adults, 200 billion red blood cells and 10billion
Hemopoietic tissue refers to the tissue
where-hemopoiesis takes place, i.e.: where
the formed elements of blood develop from
primitive cells (ste-m cells) to mature forms.
There are two kinds of hemopoietic tissue:
lymphoid tissue and myeloid tissue.

During prenatal life, blood cell formation
is initially the function of the mesoderm of the
¥olk.:sa'c.This task is soon transferred to the
liver, and then the sp:ie-en.But by the second
month ofintrauterine life,some longbones start'
to ossify and the bone marrow becomes a site
for blood cell formation. Subsequently, as more
and more bones ossify, the blood cell-forming
activity of the liver and spleen decreases.
Postnatall , the production of formed
elements of the blood is the exclusive domain
Lymphoid Tissue
In lymphoid tissue, only lymphocytes are
produced. Lymphoid tissue is discussed in the
chapter on lymphoid system.
Myeloid Tissue
In myeloid tissue, all formed elements of
blood, including lymphocytes, are produced.
From birth onwards, myeloid tissue is
synonymous with red bone marrow. In
newborns, all the cavities in practically all
the bones in the body are filled with red bone
marrow. But as the bones increase in size,
adipose tissue invades most cavities in bones.
Red bone marrow in these cavities becomes ORIGIN OF HUMAN CELLS
yellow bone marrow and ceases to be a site
for hemopoiesis. In adults, red bone marrow is The ancestry or lineage of all the cells in the
confined to the spongy portion of flat bones, body, including blood cells, can be traced back
notably the sternum and 'lium, vertebrjrl to a single cell, the fertrltzed oxurrr or y-gD.t.~.
bodies, and the upper part of the humerus and This cell results from the union of a sper.m..cell
femur. and an gg cell.

The stroma of myeloid tissue is made up The zygote is a tutipo'tent stem cell,
of reticular fibers and reticular cells. It provides meaning it can multiply infinitely and has the
the framework for a very complicated network potential to give rise to any type ofnnman cell,
placenta cell, or cell oEtoe fetal memhra-n-es)
of interconnecting spaces Emarrow c vitie:s)
(i.e., a-mnion and dlnri.a:-n that surround and
supplied with numerous sinusoids GsinuS::QidaL
protect the fetus in utero),
capillaries). These spaces are also filled with
blood cells of the 'different lineages in varying
stages of differentiation (that comprise the Stem Cells
~arenehyma) and adipocytes. Incidentally, Soon after it has formed, the zygote
in histology, the term "stroma" refers to undergoes cleavage-it divides many times
the supporting framework which is usually over. The cells, called b-Iastomeres, that
connective tissue. The ,terin "parenchyma" arise during the first few days of embryonic
refers to the functional elements of a tissue or development form a spherical structure referred
organ. to as morula because it looks like a mulberry.
The sinusoids that strew red bone marrow Like the zygote, blastomeres are totipotent stem
are lined with thin endothelial cells that rest c.~lls.Totipotent stein cells, however, start to
on a discontinuous basal lamina. Immediately differentiate shortly after they are formed.
external to the endothelial cells are some When stem cells differentiate (i.e.,
macrophages (p-erisinusoiilal mac_rop-hages~. transform to specialized cells), they make a
These are responsible for removing foreign series of commitment decisions, retaining
particulate material and worn-out red blood differentiation potential for some lineages
cells from the blood and the marrow cavities. while losing others. As cells become more
To help them phagocytose particulate material differentiated, they become progressively
from circulating blood, the perisinusoidal more restricted in their lineage potential
macrophages have processes that extend into until eventually, they become unipotent or
the lumen of the sinusoids. committed to the formation of a single cell
type. Likewise, as stem cells differentiate, their
Newly matured blood cells in the bone
capacity to multiply progressively decreases.
marrow cavities find their way into circulating
blood by transeelfular migratio across By the fourth day of intrauterine life, when
the endothelial lining of the sinusoids. the embryo has turned into a fluid-filled sphere
Particularly, when a mature blood cell presses called blcrstocfst, true totipotent stem cells do
on the endothelial cell that lines a bone marrow not exist anymore.
sinusoid, the endothelial cell membrane The blastocyst consists of two types of
responds by forming a temporary opening, cells. One cell type forms the enclosing wall or
called migration :Rore, which closes after the tronhoblast of the blastocyst. It gives rise to the
blood cell reaches the lumen of the sinusoid. placenta and the fetal membranes. The other

ESTE3/\N & CC)~~z/\LES'TEXTBOOK OF HISTOLOGY

cell type forms a cluster of cells, called Innen
c_ell mass; on one pole of the blastocyst. The
cells of the inner cell mass, otherwise referred to
as (emhr.¥omc slellLc_eUs, give rise to the fetus.
They are no longer totipotent because they can
no longer form a placenta or fetal membranes.
They are simply pluripotent and can produce
any specialized cell type of the body. Subsequent
divisions of the cells of the inner cell mass give
rise to more pluripotent stem cells. In a matter of
days, these stem cells further differentiate into
multigotent stem cells.
Multipotent stem cells are still versatile-
they can transform into numerous, albeit
finite, types of cell and replicate very fast.
Examples of multipotent stem cells are
me_sencli mal cells which, as discussed in the
Multipotent stem cells are plentiful in
the embryo and fetus but rare ip adults. In
adults, multipotent stem cells, albeit scarce, are
probably present in most organs of the body
where they replace dead or damaged cells.
Hemopoietic Stem Cell
Hemopoietic stem cells appear in.the ;y;olk.
sac as early as after two weeks of embryonic
life. However, they remain essentially
homeless throughout much of the individual's
intrauterine life. In their search for a site that
can provide the appropriate environment for
their proliferation and differentiation, the
hemopoietic stem cells and their progenies
find shelter initially in the yolk sac, then
the developing liver, then the spleen, before
settling permanently in the bone marrow.
In the organs they colonize (i.e., yolk sac,
liver, spleen, and bone marrow), the hemopoietic
stem cells proliferate either to renew their
Hemopoietic Growth Factors and
Hormones
Hemopoiesis is controlled by a number of
hormones and hormone-like growth factors.
Many of these factors act synergistically on the
different cell lineages.
The hormone-like growth factors are
produced within the bone marrow by stromal
cells (f:i6);olilasfs and r~ticular. cells:),
m_n_Do_q':tes and m-a-crl[phag~s, enilotlielial
cells, and some.jy:mp_h:oc~es. Examples of these
growth factors are stem cell factor. eSC ) which
stimulates the proliferation and subsequent
differentiation of the multipotent stem cells;
g-n-nuloq:te colony:-stimula_ting facto!!
€ -CS ) which stimulates the production
and subsequent differentiation of the stem
cells of the neutrophilic lineage; granulo€yte-
macr-OPhage culrrny:-sttrtllrIati-n-gfa-ctor~GM-
CSE} that stimulates the production, growth,
and differentiation of the cells of the basophilic
and eosinophilic lineages, but which also has a
similar effect on the cells of toe other lineages
including the erythroid, neutrophilic, and
monocytic lineages; and mon_o_c:y;te-cn0 y:-
(stimulating fador M-CSE) which stimulates
monopoiesis.
The hormones
...
f,-rytlImp-oieti'nis the principal regulator of
RBC production. It is a glycoprotein produced
by the kidneys and, to a small extent, the liver.
It promotes the differentiation of the cells of the
erythroid lineage. It also influences the more
differentiated cells in the lineage to increase
their iron uptake and hemoglobin production.
;:[.nromoopoielin, on the other hand, is the
primary regulator of megakaryopoiesis and
HEMOPOIESIS ...
platelet production. It is produced primarily
by the liver and, to a limited extent, by the
kidneys. Thrombopoietin stimulates the
production and differentiation of the cells in
the megakaryocytic lineage. Recent evidence
indicates that thrombopoietin also profoundly
influences the development of the other
hemopoietic stem cells.
Progenitor Cells
T stem cell are committed to become B-cell
and :I-cell, respectively. The DC stem cell is
committed to become a I:}':mphoid related
den"dritic cell while the myeloid stem cell is
committed to ,give rise to the other formed
elements o£]jloud. . it becomes a Colon:y:-Eorming
There are two kinds of progenitor
cells: early progenitor cells which are still
multipotent and/or have extensive proliferative
capacity, and late progenitnr c-ellswhich can
transform only to cells of a specific lineage
and have limited proliferative capacity. The
progenitor cells that have been enumerated in
the preceding paragraph are early progenitor
cells.
Myeloid Stem Cell
The mxeloid stem cellsretain their capacity
to divide to renewtheir numbers throughout the
individual's lifetime. However, this capability
diminishes as the individual ages. In adults,
myeloid stem cells comprise only about 0.2%
of the total population of nucleated cells in red
bone marrow.
ESTEBAN & Cm~Z!\LES< TEXTBOOK OF HISTOLOGY
itlnit-Granulocy:fe Macrophage (CFU-GM),
Colony-Forming Unit-Eosinophil (CFU-
Eo), Colon}':-Forming Unit-Basopliil (CFU-
Bas), and Colony-Eorming Unit-Mast Cell
(CFU-Mast). eF::t1-G:EMM,BFB-E, BEU-MW.,
and eEt1-GM are still early progenitor cells
while e_FtI-Eo, CtF.tI-Bas,and €EU~ ast are
already late progenitor cells..
The Burst-Forming Unit-Erythroid
(BFU-E) is committed to produce RBCs only,
but it still has extensive proliferative capacity.
When its progenies differentiate, each of them
becomes a e_Jd:ony:-Eorming nf -Er:y:tfiroHl
GeED-E), a late progenitor cell that has limited
proliferative capacity.
A Burst-Forming Unit-Megakaryocyte
(BFU-MK) still has extensive proliferative
capacity, but is committed to becoming
a megakaryocyte. When it differentiates,
B 't-
_egakar:f-ncyte (CFU-MK) which, like all
late progenitor cells, has limited proliferative
capacity.
A Colony-Forming Unit-Granulocyte
Macrophage (CFU-GM) is still a multipotent
cell that can give rise to two late progenitor
cells of the WBC lineages: Eolonr-Forming
Bnit-6ranuloc~te (CFU-G) that is destined
to produce neutro hils, or cColony:-Eorming
Hni -MonocyteiBen-dritic €ell (CFU-M/
DC) that is destined to produce either
monocy:tes or m:r-eJojd-relate(i(iena rific cells
(discussed in chapter on lymphoid system).
Incidentally, as mentioned in the chapter on
cartilage and bone, the CFU-GM is probably
also the stem cell for the '0 eoclasts
The Colony-Forming Unit-Eosinophil
(CFU-Eo), Colony-Forming Unit-Basophil
(CFU-Bas), and Colony-Forming Unit-Mast
Cell (CFU-Mast) are committed to produce
eosinoBhils, 6asophUs, and mast cells,
respectively.
The different stem cells and progenitor
cells of the formed elements of blood can
be distinguished from each other with the
Fig. VIII-1. Erythropoiesis. The bone
marrow smear demonstrates the
diffferent stages of erythropoiesis.
The precursor cells-proeryhtroblasts
(pro}-successively become basophilic
erythroblasts (be), polychromatophilic
erythroblasts (pe), normoblasts (no),
reticulocytes (see Fig. VIII-4), and mature
RBC (rbc). The smear also exhibits some
platelets (pi). Wright x 1,000.

use of special histologic techniques. These from which they differentiate are as follows (see
include marrow culture techniques and also Table VIII-I):
immunocytology. However) the cells are 1. e:r~throid lineage - proerytlnoblast,
morphologically indistinguishable under the which differentiates from the CFU-E
microscope) even at very high magnification.
2.
They are all small) round cells with a centrally-
located nucleus that contains fine chromatin
material and two or more nucleoli) and scanty .
and basophilic cytoplasm. and CFU-Bas) respectively
3.
Precursor Cells

When a late progenitor cell differentiates) it

gives rise to erecursor cell. This is the earliest 4.
cell form of alineage that can be morphologically
distinguished from the cells of the other lineages
5.
under the light microscope. The precursor cells
of the different lineages and the progenitor cells

Table VIII-1. Progenitor and Precursor Cells of

the Different Blood Cell Lineages
ERYT R
The proerythroblast goes through
several stages of differentiation before it
becomes a mature RBC. These stages are
distinguishable in routinely-stained bone
marrow smears. They consist of the following:
basophilic erythroblast) polychromatophilic
erythroblast) normoblast, and reticulocyte.
T stem cell It takes the proerythroblast about a week to
B-cell B stem cell differentiate into a mature RBC. '

HEMOPOIESIS ~
 In the erythroid lineage, only the Polychromatophilic Erythroblast
p-ro~eq~throblasts and oasophilic eI}7:throblasts (Polychromatophilic Normoblast;
are capable of mitosis. These two cell types
Intermediate Normoblast)
undergo a total of three to five cell divisions.
In" erythropoiesis, as a cell becomes A polychromatophilic erythroblast is,12to
more mature, it decreases in size; its nucleus
15 ~m in diameter. In routinely-stained smears,
its nucleus which occupies only about 50% of the
progressively shrinks till it becomes pyknotic,
cell is noted to contain coarse "checkerboard"
after which, itis extruded; its ribosomes decrease
chromatin, but no nucleolus. The cytoplasm
in number, leading to diminished basophilia
of a polychromatophilic erythroblast-which
of the cytoplasm; and its hemoglobin content
already -cont ains a significant amount of
increases, leading to increased cytoplasmic
hemoglobin-is grayish pink.
acidophilia.
Electron micrographs ofpolychromatophilic
erythroblasts show few organelles. I
Proerythroblast (Pronormoblast)
A pm_cr:y:thro:olastis a large cell that is 22 Normoblast (Orthochromatic
to 28 ~m in diameter. In routine bone marrow Erythroblast; Late Normoblast)
smears, its spherical and centrally-located
nucleus occupies about, 80% of the cell and A normoblast has a diameter of 9 to 11 ~m.
possesses fine chromatin granules and one It has an eccentric and pyknotic nucleus (i.e.,
the' chromatin material is highly condensed)
to two prominent nucleoli. Its cytoplasm is
that occupies less than 25% of the cell. A
scanty and basophilic, but around the nucleus,
normoblast already contains a considerable
it forms a pale area (}2erinude-arhalo~ that has
amount of hemoglobin. Thus, its cytoplasm,
been shown by electron microscopy to contain
in routinely-stained smears, is already pinkish.
mitochondria, Golgi complex, centrioles, and
Electron micrographs reveal that a normoblast
abundant ribosomes. A proerythroblast does
has paucity of organelles that include a few
not contain hemoglobin yet. ribosomes and degenerating mitochondria and
Golgi complex.
Basophilic Erythroblast (Basophilic
Normoblast; Early Normoblast) Reticulocyte (Polychromatophilic
Erythrocyte)
A 15asepliilic er::ytnm_biast is 16 to 18 ~m
in diameter. In routinely-stained smear-s, its A reticulocyte is a normoblast that has
spherical nucleus takes up about 75% of the extruded its nucleus. Its cytoplasm, however,
cell and is seen to contain chromatin that is as electron micrographs show, still possesses
coarse and arranged in a clock-face pattern, centrioles, a'few mitochondria, Golgi complex
and an occasional nucleolus. A basophilic . remnants, and some ribosomes.
erythroblast already synthesizes hemoglobin Reticulocytes still produce hemoglobin.
but its cytoplasm is still highly basophilic, The hemoglobin that a reticulocyte still needs
often more basophilic than the nucleus. to synthesize could be as much as 20% of what
Electron micrographs of basophilic it should have as a mature RBC.
erythroblasts reveal a well-developed Golgi It is not unusual for a few reticulocytes to
complex, many ribosomes and mitochondria, prematurely enter blood where they complete
and the presence of microtubules and their maturation in about 24 to 28 hours. In
microfilaments. routinely-stained blood smears, reticulocytes

ESTEBf\N & CiONZALES' TEXTBOOK OF HISTOLOGY

-
can be distinguished from mature RBCs by
their size-they are larger (9 l-tm,diameter)-
and by the bluish tinge in their cytoplasm due to
the remaining cytoplasmic organelles.
GRANULOPOIESIS
The neutrophilic, eosinophilic, and
basophilic myeloblasts go through several
stages of differentiation before they become
mature granulocytes. These stages consist of
the following: promyelocyte, myelocyte, and
metamyelocyte.
In routine bone marrow smears, the
neutrophilic, eosinophilic, and basophilic
myeloblasts look alike despite the fact that
they arose from different progenitor cells.
Nevertheless, they can be distinguished from
the precursor cells of the other cell lineages.
The same is true with the neutrophilic,
eosinophilic, and basophilic promyelocytes.
They cannot also be distinguished from each
other in routine bone marrow preparations.
Myeloblast
Myeloblasts are fairly large round cells that
have a diameter of 15 to 20 l-tm.In routine bone
marrow LM preparations, their nucleus is seen to
Fig. VIII-2. Granulopoiesis. The bone
smear illustrates many of the stages
of granulopoiesis: promyelocyte
(pro), neutrophilic myelocyte (my),
neutrophilic metamyelocyte (met),
band form (bf), and neutrophil (ne).
Wright x1,OOO.
be oval or round. The nucleus also contains one
to three nucleoli and fine chromatin granules
that are evenly dispersed, although sometimes, a
few small clumps may be seen. Their cytoplasm
is scanty, moderately basophilic, and contains
no granules.
Electron micrographs of myeloblasts reveal
an abundance of mitochondria and ribosomes
and a well-developed rER.
Myeloblasts can still divide, but their
proliferative capacity is limited. Invariably, their
progenies differentiate into promyelocytes.
Promyelocyte
In routinely-prepared bone marrow smears,
a promyelocyte is larger than a myeloblast. Its
diameter can reach 25 l-tm.Its round or oval
nucleus contains clumped chromatin material
that makes it difficult to appreciate the nucleoli.
Its cytoplasm is more basophilic than that of the
myeloblast. In addition, it contains azurophilic
granules (primary granules) which are not yet
present in the myeloblast.
Electron micrographs of the promyelocyte
confirm the presence of membrane-bound
cytoplasmic granules and a well-developed rER.
HEMOPOIESIS _
 Like a myeloblast, a promyelocyte is still
capable of mitosis.
Neutrophilic Myelocyte,
Metamyelocyte, and Stab Cell
When promyelocytes acquire specific
granules (secondary granules) in their
cytoplasm, they become myelocytes. From
this point on, the cells of the neutrophilic,
eosinophilic, and basophilic lineages can be
distinguished from each other in routinely-
prepared bone marrow smears.
A neutrophilic myelocyte is slightly smaller
than a promyelocyte. Its nucleus is usually ovoid
and contains coarse chromatin material that
forms clumps but has no visible nucleolus. Its
cytoplasm is more abundant but less basophilic
than that of a promyelocyte. As in mature
neutrophils, its specific cytoplasmic granules
have little affinity for dyes and are too small
to be appreciated under the light microscope.
In routine bone marrow smears, however, they
impart a lilac hue to the cytoplasm. They first
appear near the nucleus. As they increase in
number, they begin to be seen in the periphery
of the cell.
A neutrophilic myelocyte has also
azurophilic granules which, in electron
micrographs, are denser than the specific
granules.
A neutrophilic myelocyte undergoes up
to three cell divisions before its progenies
differentiate into neutrophilic metamyelocytes.
A neutrophilic metamyelocyte is already
about the size of a mature neutrophil and no
longer capable of mitosis. Its nucleus is deeply
indented on one side and contains chromatin
granules that form coarse, dark clumps. Its
cytoplasm is slightly basophilic. The specific
granules-which could already be equal to the
normal mature complement-far outnumber
the azurophilic granules.
A neutrophilic metamyelocyte
differentiates into a stab cell or band form. In
routine bone marrow smears, the cytoplasm of
ESTEBAN8< CiONZALES'TEXTBOOK OF HISTOLOGY
a stab cell is pale violet and already contains the
mature complement of specific granules.
A stab cell can be distinguished from a
neutrophilic metamyelocyte by the appearance
of its nucleus, often U- or S-shaped or elongated
and slightly curved. As mentioned in the
previous chapter, some stab cells are normally
present in blood. When the nucleus of the stab
cell has formed lobes, it has transformed into a
mature neutrophil or segmenter.
It takes a neutrophilic myeloblast about 11
days to transform into a segmenter. During this
period, the differentiating cell undergoes five
mitotic divisions.
Eosinophilic Myelocyte and
Metamyelocyte
As in the neutrophilic lineage, the
first cell in the eosinophilic lineage that is
morphologically distinguishable from its
granulocytic counterparts in routine bone
~~rrow smears is the myelocyte.
An eosinophilic myelocyte has a lot in
common with a neutrophilic myelocyte in
routine bone marrow smears. Its chromatin
material forms coarse clumps and its cytoplasm
is slightly basophilic and contains azurophilic
granules. The feature of an eosinophilic
myelocyte that distinguishes it from a myelocyte
of the neutrophilic and basophilic lineages is its
specific cytoplasmic granules. These granules
are acidophilic and much larger than those in a
neutrophilic myelocyte.
An eosinophilic myelocyte differentiates
into an eosinophilic metamyelocyte-which
looks like its neutrophilic counterpart-except,
again, for its distinctive specific cytoplasmic
granules.
Unlike in the neutrophilic lineage, there is
no band form in the eosinophilic lineage. When
the nucleus of a metamyelocyte has formed
lobes (usually two, but occasionally three), the
cell has differentiated into a mature eosinophil.
-- ---------------
Basophilic Myelocyte and
Metamyelocyte
In routine bone marrow smears, the
basophilic myelocyte and metamyelocyte
look like their neutrophilic and eosinophilic
counterparts except for their specific
cytoplasmic granules and their nucleus that
stains less intensely and has finer chromatin
granules. The specific cytoplasmic granules
of the basophilic myelocyte and basophilic
metamyelocyte have a high affinity for basic
dyes. They are blue or dark purple in routinely-
stained prepa~ations. Furthermore, they are
larger, fewer, and of variable sizes compared
with those in their eosinophilic counterparts.
Like in the eosinophilic lineage, there is
no band form in the basophilic lineage. The
metamyelocyte differentiates directly into a
mature basophil.
THROMBO 01
Platelets are cytoplasmic fragments:'
of a giant cell in the bone marrow called
megakar.:yocrte. Thus, thrombopoiesis
encompasses 1) the development of a
megakaryocyte (megakarropoiesiSJ); and
Fig. VIII-3. Bone Marrow Smear.
The section shows several types
of bone marrow cells including a
megakaryocyte (me), plasma cell
(pc), eosinophilic myelocyte (emy),
eosinophilic metamyelocyte (eme),
eosinophil (eo), and reticulocyte (re).
Wright x1,OOO.
2) the process of forming platelets from a
megakaryocyte.
Megakaryopoiesis
A egakar¥oolast, the precursor cell of the
megakaryocytic lineage, is about 15 to SO[lm
in diameter. In routine bone marrow smears, it
has a large, slightly indented, ovoid nucleus that
contains loose chromatin material and multiple
but inconspicuous nucleoli. Its homogeneous
cytoplasm is intensely basophilic because of the
presence of numerous ribosomes. In electron
micrographs, aside from ribosomes, the
megakaryoblast is seen to also contain many
mitochondria, a well-developed Golgi complex,
andanrER.
A megakaryoblast undergoes a series (up
to seven) of incomplete type of mitosis called
endomitosis. It results in the formation of a
huge cell that contains a single but multilobed
nucleus. Each time a megakaryoblast undergoes
endomitosis, its cytoplasmic elements duplicate.
However, there is no cytoplasmic division, and
its daughter nuclei remain fused.
A megakaryocyte is a giant cell (SO to
150 um). In routine bone marrow smears, its
multilobed nucleus is seen to contain coarse
HEMOPOIESIS ..
chromatin and indistinct nucleoli. Its cytoplasm
is abundant and basophilic but the basophilia
is less than that of a megakaryoblast. It also
exhibits numerous fine azurophilic cytoplasmic
granules.
In electron micrographs, the cytoplasm
a megakaryocyte is noted to have many free
ribosomes, a poorly-developed ER, several Golgi
complexes, many membrane-bound granules,
and numerous smooth-surfaced membranes
that tend to flatten.
Some authors describe a cell, which they
call promegakaryocyte, that represents an
intermediate stage between the megakaryoblast
and the adult megakaryocyte. The
promegakaryocyte is 30 to 50 ~m in size. In
routine bone marrow smears, its nucleus shows
several lobes while its cytoplasm contains
several pairs of centriol~s and fine azurophilic
cytoplasmic granules.
It takes about 10 days for a megakaryoblast
to transform into a mature megakaryocyte.
Formation of Platelets from
Megakaryocytes
Platelets are formed from megakaryocytes
in the bone marrow when the cell forms long
cytoplasm processes called pseudopodia
(platelet ribbons; proplatelets). These enter
the lumen of the sinusoids where they fragment
into platelets that then join circulating blood.
New evidence indicates anothe~ way
by which platelets are formed. Evidently,
megakaryocytes sometimes enter the bone
marrow sinusoids intact then migrate to the
pulmonary blood vessels where their cytoplasm .
fragments into platelets.
A megakaryocyte produces 4,000 to 8,000
platelets, after which, it presumably degenerates.
MONOPOIESIS
A monoblast, the precursor cell of the
monopoietic lineage, differentiates into a
ESTEBAhi & GCf--J//\LLS' TEXTBOOK OF HISTOLOGY
promonocyte before it transforms into a
mature monocyte.
Monoblast
In routine bone marrow smears, the
of
monoblast is a large round cell that is about
15 to 20 ~m in diameter. Its nucleus is oval or
round and has fine chromatin granules that are
evenly dispersed, although a few small clumps
of chromatin may be seen. Its scanty cytoplasm
is moderately basophilic.
The monoblast resembles the myeloblast
(the precursor cell of the granulocytic lineages)
in routine LM preparations. The two cells
become distinguishable from each other only
when the nucleus of a monoblast begins to
indent and its cytoplasm starts to manifest
azurophilic granules~ things that do not occur
in the myeloblast.
Electron micrographs of the monoblast
reveal free ribosomes, polyribosomes, a rER,
and a few mitochondria in its cytoplasm.
Further division and differentiation of the
monoblast give rise to promonocytes.
Promonocyte
A promonocyte is larger than a monoblast.
In routine bone marrow smears, it exhibits
a nucleus that is large, indented, and paler-
staining than that of a monoblast because its
chromatin granules are finer. Also, owing to the
fine chromatin material of the nucleus, nucleoli
are visible. Its cytoplasm is paler than that of a
monoblast but still basophilic. A promonocyte
contains azurophilic cytoplasmic granules.
A promonocyte divides twice before its
progenies differentiate into mature monocytes.
Monocytes enter blood soon after they are
formed. They stay in blood for a while before
migrating into connective tissue where they
transform into macrophages.
It takes monoblasts about 55 hours to
transform into monocytes.
LYMPHOPOIESIS The B-stem cells, on the other hand, start
to differentiate in the bone marrow when they
Lymphopoiesis occurs in the various receive the appropriate signals in the form of
lymphoid tissues and lymphoid organs in the growth factors. Immunocytology has shown
bO,dy.These tissues and organs are classified that they pass through two intermediate stages
into two groups: central lymphoid organs (Pro-B cell and Pre-B cell) before they become
which refer to the thymus and bone marrow; B-Iymphoblasts, the ~recursor cells of the
and peripheral lymphoid tissues and organs lineage. B-Iymphoblasts proliferate extensively.
which refer to the lymph nodes, spleen, and Subsequently, their progenies differentiate into
the mucosa-associated lymphoid tissues B-prolymphocytes before becoming mature
(MALT). MALT include the tonsils and the B-cells.
other nonencapsulated lymphoid tissues in
the gastrointestinal (i.e., GALT),respiratory Mature T- and B-cells leave the central
(i.e., BALT),and genitourinary tracts. lymphoid organs by entering blood or lymphatic
vessels. They then migrate to, and take up
residence in, the various peripheral lymphoid
Lymphopoiesis in the Central
tissues and organs.
Lymphoid Organs
The mature T- and B-cells that leave the
The thymus and the bone marrow are thymus and bone marrow, respectively, are
called central lymphoid -organs because these immunocompetent but still naive (i.e.,they have
are where the progenitor cells of lymphocytes, not encountered an antigen yet.)
i.e., T and B stem cells, develop into mature and
In routine light microscopic smears, the
immunocompetent T- and B-cells, respectively.
B-Iymphoblast and T-Iymphoblast look alike.
As soon as they have differentiated from . The same is true for the B-prolymphocyte and
hemopoietic stem cells in the bone marrow, T-prolymphocyte.
the T-stem cells migrate to the thymus while
the B-stem cells stay in myeloid t issue (i.e., Lymphoblast
bone marrow). Incidentally, hemopoietic stem
cells-from which the B-stem cells and T-stem A lymphoblast is a large cell that is 2S
cells are derived -abound in the bone marrow ~m or more in diameter. In routine histologic
of embryos and fetuses. However, their numbers preparations, it is seen as having relatively
diminish rapidly with age such that in adults, abundant basophilic cytoplasm (nucleus to
relatively few still exist. Nevertheless, the bone cytoplasm ratio is about 4:1) that does not
marrow remains a source of T-stem cells and contain azurophilic granules. Its nucleus is
B-stem cells throughout life. large and spherical. It contains fine and highly
dispersed chromatin material and one or two
The exact origin of the NK cells is not
easily visible nucleoli.
known yet. They probably originate from the
bone marrow but definitely not from B- or T- Normally, lymphoblasts do not circulate
stem cells. They likewise do not get processed in blood; they proliferate and differentiate into
in the thymus. prolymphocytes in the central lymphoid organs.
In the thymus, the T-stem cells
differentiate into 0::- =r-mplio:bb-ns (the Prolymphocyte
precursor cells of the lineage) and proliferate. A prolymphocyte is slightly smaller than
Subsequently, T-Iymphoblasts transform into a lymphoblast. In routine smears, its nucleus
T-prolymphocytes before becoming mature shows condensed chromatin material that
T-cells. makes nucleoli hard to appreciate. Its cytoplasm

HEMOPOIESIS _
is basophilic and already contains azurophilic
granules.
Further differentiation of a prolymphocyte
gives ~ise to a mature lymphocyte.
Lymphopoiesis in the Peripheral
Lymphoid Organs
When they leave the central lymphoid
organs and settle in the peripheral lymphoid
tissues and organs, the B- and T-cells are small
lymphocytes that retain their mitotic capability.
This capability is aroused if they get activated,
which occurs when they react to an antigen.
An antigen is any substance perceived by the
cells of the lymphoid system as foreign to the
body. The presence of an antigen in an area
of the body activates the lymphocytes that
are in the lymphoid tissues-in and around the
area. (Further discussion on the activation of
lymphocytes is presented in the chapter on
lymphoid system.)
When activated, lymphocytes become
.from one stage to the next is a very gradual
larger.Thereafter, they divide extensivelybefore
differentiating into the different functional
types oflymphocytes.
Some activated lymphocytes leave the area morphological gradations between stages is
of antigen exposure and enter the bloodstream infinite.)
EsrEBAN & GONz/\LES' TEXTBOOK OF HISTOLOGY
(a possible reason why there are large
lymphocytes in blood). Then they seed the
other peripheral lymphoid tissues and organs
in the body where they likewise proliferate
extensively before differentiating into various
functional types.
Re-circulating Pool of Lymphocytes
Lymphocytes, regardless of where they are
produced (i.e., central or peripheral lymphoid
tissues), do not settle permanently in any
peripheral lymphoid tissue or organ. They
shuttle back and forth between the different
lymphoid tissues and organs by joining blood
or lymph to comprise a "re-circulating gool."
The T-cellsre-circulate more often than the
B-cells.
(Note: Figure VIII-4 on the next page
shows the various stages that the stem cells of
the formed elements of blood undergo before
becoming mature cells. One must, however,
bear in mind that the transformation of a cell
process. Often, changes in the cytoplasm
do not go hand in hand with changes in the
nucleus. Accordingly, the number of possible
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HEMOPOIESIS
 ,f
~'
he skin or in egu ~e~nt is the tough
T protectiv ,external covering of the bod
that is continuous with the mucos~J
(mucous m!mbrin~ej that lines the mouth,
nose, anal canal, genital openings, and eyes.
It is the largest organ of the body in terms of
weight, accounting for about 15% of a person's
body weight. Its surface area in adults exceeds
1.5 rrr'. Its thickness ranges from 0.5 to S.Om
depending on its location. It is thinnest overthe
eye ia.sand thickest over the sol(!slfof tile'feet.
The skin'sprotective functio is not limited
to preventing harmful substances from entering
the body; it alsoincludes preventing loss ofbody
fluids. Aside from protecting the other tissues
of t e body, the skin, by forming sweat, helps
regulate body temperature, and excrete w:aste
materia s and waste products. It also serves as
a huge sense organ that is receptive to touch,
pain, pressure, and temperature. Furthermore,
some of its cells assist in the immune response
(immune defense) of the body. The skin is also a
siteforthe production 0 itarn...!i~1Which occurs
when 1-dehyd 01::Jii'_0 €st'e,@rf'hatis present in its
epithelial coverreacts with ultraviolet light from
the sun.
Two properties of the skin are occasionally
taken advantage of by medical science. Firstly,
the skin, being a semipermeable membrane,
can absorb certain substances from its surface,
_I,
Skin and Its
Appendages
hence, some pharmaceutical preparations are
specifically designed for absorption through
the skin. Secondly, in the finger tip , the skin
exhibits shallow grooves and ridge whose
J
patterns are unique for each perso . These
grooves and ridges, which are collectively
referred to as fingeF:er·nl\.s',
are used as basis for
individual identification.
Associated with the skin are several
structures, collectively referred to as
appendages'of"th~esltin. The appendages of
the skin consist of nail, hair, and cutaneous
glands-sweat and sebaceous glands.
The skin and its appendages are often
collectively referred to as the integumentiiiyJ
LAYERS OF THE SKIN
Histologically, the skin consists of two
distinct layers: epidermis and dermis. The
epidermis, the more superficial layer, is an
epithelial coat derived from ectoderm while
the dermis (corium) is made up of connective
tissue derived from mesoderm.
The junction of the epidermis and dermis
is well-delineated, but not smooth. Tissue
elements of the dermis form irregular conical
projections called dermal papillae into the
epidermis. Conversely, irregular conical
invaginations of the epidermis called epidermal
epidermI.
ridges (rete pegs; epidermal pegs) invade the
dermis.
The dermis is anchored to the underlying
tissues by loose connective tissue that is referred
to as hypodermis or subcutaneous tissue that
often contains numerous adipose cells.

Epidermis
The epidermis is a keratinized stratified
squamous epithelium. Its most superficiallayer
is continuously shed such that the epidermis is
completely renewed every 20 to 30 days. This
renewal process is the responsibility of the
keratinocytes, the principal cells of the layer.

Keratinocytes
Keratinocytes comprise 85%to 95% of the
Fig.IX-1. Layers of the Skin.The photomicrograph
cell population of the 'epidermis. They are cells
demonstrates the two histologic layers of the
that are specialized to produce keratin-an skin-the epidermis which is an epithelium and
intermediate filament that is also present but the dermis made up of connective tissue-and
in much lesser amounts in other epithelial cells. the hypodermis which binds the skin to the
underlying tissue. Thin Skin, H&E x 100.
The keratinocytes in the deep layers of the' .
epidermis divide continuously either to renew and sebaceous glands while thick skin contains
their numbers or to differentiate and keratinize numerous sweat glands, but lacks hair follicles
(i.e., produce and accu~ulate keratin in their and sebaceous glands.
cytoplasm). As differentiated keratinocytes
grow older, they are slowly pushed towards Thick Skin
the surface of the epithelium by newly-formed
Thick skin covers the palms and. soles.
and younger cells. At the same time, they
Its epidermis consists of five layers. From the
accumulate increasing amounts of kera in and
deepest to the most superficial, these layers
become bigger.When they are already near the
are the 1) stratum germinativum (stratum
surface, they dry up, become smaller then die,
basale), 2) stratum spino sum (prickle cell
and ultimately, get shed off.
layer); 3) stratum granulosum; 4) stratum
lucidum (clear layer); and 5) stratum corneum
Types of Skin (hornycell layer}. The stratum corneum and
Skin is categorized into two types: thick stratum lucidum are sometimes collectively
and thin. referred to asthe cornified layer while the three
The main basis for classifying skin into other layers comprise the stratum Malpighii.
thick or thin is the thickness of its epidermis,
Stratum Germinativum (Stratum Basale)
but thick and thin skins differ in a few other
aspects. The dermal papillae in thin skin are The stratum germinativum consists mainly
shorter than those in thick skin. Furthermore, of a single layer of tall cuboidal keratinocytes
thin skin contains sweat glands, hair follicles, that rest on a basement membrane.

SKIN AND ITS APPENDAGES "*'

 The keratinocytes in the stratum
germinativum have a large nucleus and
basophilic cytoplasm that contains numerous
ribosomes, a Golgi complex, endoplasmic
reticulum, a limited number of mitochondria,
and a few keratin filaments. They are bound on
their lateral and upper surfaces to neighboring
cells by desmosomes and to the basal lamina by
hemidesmosomes.
In electron micrographs, many keratin
filaments are noted to converge and insert
into each half-disc of the desmosomes in
keratinocytes.
The keratinocytes in the stratum
germinativum divide continuously: hence, in
routine histologic preparations, many mitotic
figures are seen among the cells.
Stratum Spinosum (Prickle Cell Layer)
The stratum spinosum consists mainly
of polyhedral keratinocytes that are arranged
into several layers. These cells are shorter and
less intensely basophilic in routine histologic
preparations than those in the stratum
germinativum. Their cytoplasm contains
the same organelles as those in the stratum
germinativum, but their keratin filaments,
which are more numerous, form bundles.
The keratinocytes in the stratum spinosum
are still tightly bound to their neighbors by
desmosomes. Incidentally, desmosomes are
Fig. IX-2. Thick Skin. H&E x1 00.
l SrT_Bf\\!& GO\Iz/\L.ES' TEXTBOOK OF HISTOLOGY
the only cell-to-cell junctions that exist among
the cells of the epidermis. Under the light
microscope, because of shrinkage caused by
routine preparations, the desmosomes appear
like spines or hairs that protrude out of the
cells. These spines, the reason why the layer is
called stratum spinosum, are often referred to
as "intercellular bridges." They are are merely
preparation artifacts.
The keratinocytes in the deeper layers
of the stratum spino sum still possess limited
capacity to mitose.
Stratum Granulosum
The stratum granulosum consists of 3 to 5
layers of keratinocytes that are more flattened
than those in the stratum spinosum. The
keratinocytes have a pyknotic nucleus and
degenerated cytoplasmic organelles.
The most distinctive feature of the
keratinocytes in this layer is the presence of
numerous dark, basophilic keratohyaline
granules in their cytoplasm. The keratohyaline
granules are not membrane-bound, and, despite
their name, do not contain any keratin. The
keratin filaments in the cytoplasm either pass
through or get attached to the periphery ofthese
granules. The keratohyaline granules contain
a histidine-rich protein called profillagrin,
which, when the cells reach the stratum
corneum, becomes fillagrin that provides the
interfilamentous matrix for the aggregation of
keratin filaments.
Another feature of the cells in this layer
is the presence of membrane-bound, lipid-
containing secretory granules (lamellar
granules) in the cytoplasm. These granules
.are also present, but to a limited degree, in the
keratinocytes ofthe stratum spinosum. They are
called lamellar granules because they consist of
closely-packedparallellamellae.The contents of
the lamellar granules are secreted and deposited
into the intercellular spaces. Together with the
desmosomes, they form an important barrier
against the entry or exit of substances to and
from the body.
 Thin Skin
Thin skin covers the whole body except
the palms and soles. Its epidermis differs from
that of thick skin in terms of the following: its
stratum spinosum is much thinner; 'its stratum
granulosum is poorly developed, and sometimes,
even absent; it does not have a stratum lucidum;
and, its stratum corneum is much thinner. Thus,
often, the epidermis of thin skin has only three
layers: stratum basale, stratum lucidum, and
stratum corneum.

Other Cell Types in the Epidermis

Aside from keratinocytes, three other
types of cells are present in the epidermis-
Fig. IX-3. Epidermis of Thick Skin. The epidermis melanocytes, Langerhans cells, and Merkel
of thick skin has five layers. From the deepest to cells.
the most superficial, these are: stratum basale
(b), stratum spinosum (s),.stratum granulosum (g),
stratum lucidum (I), and stratum corneum (c). H&E
Melanocytes
x400. Melanocytes are cells that are specialized to
produce melanin pigment. They are present in
The keratinocytes in the stratum
certain parts of the body, notably the skin and
granulosum can no longer divide. In fact, they
the uvea in the eye. In the skin, they comprise 7%
are already dead cells.
to 10% of the cell population of the epidermis,
which makes them second to the keratinocytes
Stratum Lucidum (Clear Layer)
in terms of number. Some melanocytes are also
The stratum lucidum consists mainly of 4 present in the superficial region of the dermis.
to 6 layers of flat, dead, anucleate keratinocytes
The melanocytes in the epidermis are
that form a light-staining, relatively translucent,
dispersed among the cells of the stratum
discontinuous layer superficial to the stratum
granulosum. The keratinocytes in this layer are
already devoid of organelles, but their keratin
filaments form thick bundles that are parallel to
the skin surface. They are likewise still bound to
their neighbors by desmosomes.

Stratum Corneum (Horny Cell Layer)

The stratum corneum consists of IS to
20 rows of flattened keratinocytes whose
cytoplasm consists entirely of keratin filaments
that are embedded in an amorphous matrix.
The deeper cells are still attached to each other,
Fig. IX-4. Epidermis of Thin Skin. In contrast to
albeit rather loosely, by desmosomes, but those
thick skin which has five layers, the epidermis of
in the superficial layers, which comprise the thin skin often has only three-stratum basale (b),
stratum dysjunctum, are free of each other and stratum spinosum (s), and stratum corneum (c).
get continuously shed off or desquamated. H&E x100.

SKIN AND ITS APPENDAGES ~


germinativum and stratum spino sum. They
are not bound to the neighboring cells by
desmosomes, but those in the stratum basale are
bound to the basal lamina by hemidesmosomes.
In H&E preparations, melanocytes appear
as "clear" cells that are difficult to tell apart from
the other clear cells (i.e., Langerhans cells and
Merkel cells) in the skin. However, they can be
distinguished with the aid of special methods
such as the DOPA technique. When incubated
in a solution of 3, 4-dihydroxyphenylalanine
(DOPA), the highly-branched processes of the
melanocytes are selectively blackened and the
cells become recognizable.
Melanocytes are small cells with long
but thin processes that are in contact with
neighboring keratinocytes. Their cytoplasm
contains a fair share of organelles and many
electron-dense, membrane-bound granules
called melanosomes. Melanin is produced
within the melanosomes that is why many
authors consider melanosomes as cytoplasmic
organelles that are unique to melanocytes.
In the melanosomes, melanin is formed
from the amino acid tyrosine through a series
of enzyme-catalyzed chemical reactions.
The melanin that a
melanocyte produces is
transferred (actually, it is the fully-formed
melanosome that is transferred) in a continuing
manner, via the melanocyte's processes, to the
keratinocytes with which it is in contact. In
the keratinocytes, the melanosomes become
permanent inclusions and are positioned in
the supranuclear portion of the cells in order
to protect the cells from solar radiation.
The transfer process of melanosomes from
melanocyte's to keratinocytes is -still a poorly
unde~~toJ)d process, but it is so efficient that,
in general,'keratinocytes contain more melanin
than melanocytes.
Each melanocyte supplies up to 30
keratinocytes with melanin. A melanocyte and
the keratinocytes that it supplies with melanin
are collectively referred to as an epidermal
melanin unit.
ESTEBAN & CCNZALES' TEXTBOOK OF HISTOLOGY
--1
I
Incidentally, skin color is an admixture
of three pigments: carotene, a substance that
is present in the intercellular substance of
the epidermis that gives the skin a yellowish
hue; hemoglobin, the red, oxygen-carrying
pigment present in the RBCs that circulate
in the capillaries of the dermis; and melanin
that gives skin its brown to black shade. The
difference in skin color among the different
races is not due to the varying number of
melanocytes in the skin, but rather to the
amount of melanin that the melanocytes
produc-e (ie., the degree of activity of the
melanocytes) .
Melanocytes are cells that arise from the
neural crest. They colonize the epidermis
during the early embryonic period. They retain
their capacity to divide throughout postnatal
life but become less mitotically active as the
person ages.
Langerhans Cells
Langerhans cells are antigen-presenting
cells (APCs)-see the chapter on the
lymphoid system-that are present not only in
the skin but also in other stratified squamous
epithelia such as those in the oral cavity,
esophagus, and vagina, In the skin, they are
mostly in the epidermis where they comprise
3% to 8% of the cells, but a limited number are
scattered in the dermis.
In the epidermis, Langerhans cells are
most numerous in the stratum spino sum.
In H&E preparations, they look similar to
melanocytes with their dark nucleus and pale-
staining cytoplasm. But with the use of special
techniques such as staining with gold chloride,
the dendritic nature of their processes become
apparent and they become distinguishable.
In electron micrographs, Langerhans cells
in the epidermis are easy to identify because
they lack desmosomes and keratin filaments.
Also, they contain rod-shaped, membrane-
bound granules called Birbeck granules
or vermiform granules whose function is
undetermined yet.
 In the embryo, Langerhans cells are
derived from the bone marrow. They colonize
the skin as early as the 5th to the 6th week of
embryonic life. Postnatally, they retain some
mitotic activity, but they renew their numbers
mainly by migration of precursor cells from
the bone marrow via blood to the skin where the
precursor cells then differentiate. In the blood,
the precursor cells probably look like and are
misidentified as monocytes.
Merkel Cells
Merkel cells are the least in number among
the cell types in the epidermis. They usually
occur singly, and occasionally in clusters in the
basal region of the epidermis in all parts of the
body, but they are most numerous in the palms
and soles.
In routine histologic preparations, they
also appear as "clear cells," like melanocytes
and Langerhans cells, and are distinguishable
under the light microscope only if specific
immunehistochemical techniques are applied.
Merkel cells are disc-shaped cells with short·
cytoplasmic processes. They are bound to
neighboring keratinocytes by desmosomes. In
electron micrographs, their cytoplasm is shown
to be devoid of keratin filaments but it has
small, dense granules that-some investigators
believe-contain a hormone similar to the
cat~ch~lamines secreted by the adrenal medulla.
A Merkel cell together" ~ith ,'an axon
termination of.a sensory neuron .forrns a
Merkel disc-a' sensory mechanical receptor
that responds to pressure and touch. As a rule,
a si4}-gleafferent nerve fiber (axon), which
generally has num~rous terminations, makes
cO,ntactwith maIl,X"Merkel cells.
.The origin cif Me~kel cells has not been
established yet, but the_prevailing view is that
they are modified Reratinocytes.
Dermis (Corium)
The dermis is much thicker than the
epidermis. Its thickness ranges from 0.6 mm
(in the eyelid) to 4.0 mm (in the back). It is
composed of connective tissue that forms two
histologic layers: papillary and reticular.
Papillary Layer
The papillary layer is the more superficial
of the two layers of the dermis. It is made up of
loose connective tissue and is well-demarcated
from the epidermis by the latter's basement
membrane.
The bulk of the connective tissue that
comprises the papillary layer form conical
projections into the epidermis called dermal
papillae. Dermal papillae are more numerous
in areas that are subjected to great pressure
such as the soles of the feet. Their core contains
a capillary bed that supplies the overlying
epidermis with blood and nerve endings
including encapsulated ones like Meissner's
corpuscles.
Reticular Layer
The reticular layer of the dermis is thicker
than the papillary layer. It is responsible for the
toughness and strength of skin. It consists of
dense irregular connective tissue that is richly
supplied with blood vessels and whose collagen
fibers form coarse bundles that are mostly
oriented parallel to the skin. Dispersed within
the reticular layer is a network of elastic fibers
that are thicker than those in the papillary layer.
Sweat glands, sebaceous glands, and hair
follicles, when present, are embedded in the
connective tissue of the reticular layer. Smooth
muscle cells are also present in this layer of
the dermis. Those that are associated with
hair follicles form bundles called arrector pili
muscles. In areas such as the scrotum in males
and areola of the breast in females, the smooth
muscle fibers form interlacing bundles in the
deep part of the reticular layer.
In the head and neck, connective tissue
bands in the reticular layer serve as insertions
of skeletal muscles that comprise the muscles of
facial expression.
SKIN AND ITS APPENDAGES ~
 The undersurface of the reticular layer of
the dermis merges with the hypodermis.
Hypodermis (Subcutaneous
Tissue)
Hypodermis refersto the loose connective
tissue that binds the dermis of the skin to the
underlying structures. In many areas of the
body (e.g., dorsum of the hand), it also allows
the skin to slide over the underlying structures.
It is not part of the skin.
The hypodermis contains numerous
adipose cells except over the eyelids, penis,
scrotum, nipple, and areola, where the
subcutaneous tissue is notably devoid of these
cells. In individuals who are over-nourished,
the adipose cells in the hypodermis over
certain areas of the body -(e.g" belly) are
particularly numerous, and they form layers
of adipose tissue called panniculus adiposus.
Sebaceous and sweat glands which are, as a
rule, confined to the dermis, sometimes extend
to the hypodermis. Hair follicles also often
invade the hypodermis.
APPENDAGES OF THE SKIN
The appendages of the skin are structures
that are derived from the epidermis. They
have specific functions and are mostly confined
to the dermis. They consist of hair, nail, and
cutaneous glands, namely the sebaceous
glands and sweat glands.
Hair
Hair is a filamentous, keratinized structure
that covers practically the whole body except
the lips, palms, soles, distal dorsal parts of
the fingers and toes, prepuce and glans of the
penis, labia minora and inner surface of the
labia majora, clitoris, and nipple. Hair varies
in thickness, length, and growth pattern in the
different regions of the body. Most parts of the
body are covered by fine, short, colorless, and
often, hardly noticeable hair. Hair, however, is
particularly lush on the scalp and the bearded
areas of the face and neck. In humans, unlike
in lower forms of animals, hair has hardly any
protective function, but it is still important to
the sense of touch.

Nerve Endings in Skin

One of the main functions of skin is to
receive and transmit sensory stimuli to the
central nervous system. Hence, the skin has a
variety of sensory nerve endings. The sensory
nerve endings consists of three groups:
simple nerve endings (free nerve endings),
expanded-tip nerve endings (i.e., Merkel
discs): and, encapsulated nerve endings
including Ruffini's corpuscles, end bulbs
of Krause, Vater-Pacinian corpuscles, and
Meissner's corpuscles. These nerve endings
have been described previously. L' #. ~
Aside from sensory nerve endings, the .: '\-~,<..~ ~";>:;::'. \11'.....
skin is also well-supplied with efferent nerve -, ~,. Jr" _ .\ 'It

....~ ... \ ,t.f. ....".... -\

fibers that regulate the activity of its glands +, ...~,\: ~~....:
) "

and smooth muscle cells, including those in the

blood vessels. Fig. IX-S. Hair Follicle. Scalp Skin, H&E x 100.

EST[BN--J & C30NZ!\LES'TEXTBOOK OF HISTOLOGY


The part of a hair that projects from the
surface of the skin is called shaft while the part
that is embedded in the skin is called root. The
hair roots are implanted obliquely, with respect
to the skin surface.
Hair Follicle
Each hair root is enclosed by two epithelial
sheaths (external and internal root sheaths). A
hair root and its sheaths comprise a hair follicle.
Associated with each hair follicle are one or
more sebaceous glands whose ducts <?peninto
the upper third of the follicle. Also associated
with a hair follicle is the arrector pill muscle, a
thin smooth muscle whose one end is attached
to the connective tissue surrounding the hair
follicle at about its mid-length. This muscle
courses close to the sebaceous gland that is
associated with the hair follicle before it attaches
its other end to the dermis. Contraction of the
arrector pili muscle straightens the hair and
forces out the secretions from the acini of the
sebaceous gland (see page l34). Simultaneous
contraction of numerous arrector pili muscles
results in "goosepimple" or "gooseflesh."
Fig. IX-6. Scalp Skin. The
section from the scalp shows
some of the appendages of
the skin. H&E x40.
In actively growing hair, the hair follicle
has an expanded, bulbous proximal portion
called hair bulb. The base of the hair bulb
has a deep concavity that is occupied by a hair
papilla which consists of connective tissue
where capillaries that supply the hair follicle
with nutrients and oxygen are embedded. The
hair papilla has an inductive effect on the cells
that produce hair and is the lifeline of the hair
follicle. The epithelial cells in hair bulb that
are associated with the hair papilla form the
hair matrix (hair root, to some authors) that
envelops the papilla.
Hair Formation and Growth
The formation of hair is similar to the cell
renewal system of the epidermis that is anchored
on the mitotic activity of the cells in the stratum
germinativum. In hair, the counterparts of
the cells in the stratum germinativum of the
epidermis are the cells of the hair matrix: they
proliferate and get pushed upward as they
differentiate, degenerate, and die. Like the cells
of the stratum germinativum, the hair-forming
cells also produce keratin, but the keratin they
produce is hard keratin. Hence, when the cells
SKIN AND ITS APPENDAGES 'Ii
die, they are not shed off; rather, they stick to
each other and string out to form hair.
Scattered among the cells of the hair
matrix are melanocytes, the cells responsible
for the color of hair. The melanocytes, like
their counterparts in the epidermis, produce
melanin, which they transfer to the other cells
of the hair matrix. ' differentiate, they transform into flattened cells
Unlike in the epidermis where the
differentiation of cells from the stratum
germinativum gives rise to just one cell type
that ultimately ends up in the cornified layer,
the cells in the hair matrix differentiate into
several cell types that occupy well-delineated
areas. Hair is the product of the three cell types
that occupy the central region of the hair matrix.
These three cell types vary in their morphology
and each type produces a different kind of
keratin. Consequently, mature hair has three
distinct concentric histologic layers: medulla,
cortex and cuticle.
The medulla, which is absent in thin
hair, occupies the core of hair. It is formed by
the cells of the hair matrix that immediately
surround the apex of the dermal papilla. When
these cells differentiate, they transform into
large, vacuolated cells that contain little keratin
filaments. Later, they degenerate and die, thus,
the medulla of mature hair consists of dead and
disintegrating vacuolated cells.
The cortex surrounds the medulla and is
the thickest of the concentric layers of hair. It
is formed by the cells in the hair matrix that
externally surround the medulla-producing
cells. When these cells differentiate, they
transform into fusiform cells that synthesize
a large amount of keratin filaments. They also
produce numerous trichohyalin granules that
contain an insoluble protein, trichohyalin.
Trichohyalin, like fillagrin that is produced
by the cells of the stratum granulosum in
the epidermis, is an intermediate filament-
associated protein that serves to aggregate
keratin filament bundles. The cells of the cortex
also receive a greater number of melanosomes
from the melanocytes than the other cellsin the
ESTE8!\~~& CiONZALES'TEXTBOOK OF HISTOLOGY
hair matrix. In mature hair, the cortex consists
of packed dead and keratinized cells that are
loaded with melanosomes.
The cuticle is the thinnest of the three
concentric layers of hair. It is formed by the
cells in the hair matrix that externally surround
the cortex-producing cells..When these cells
that synthesize a lot of keratin. Thus, in mature
hair, the cuticle consists ofseverallayersof dead,
flattened but heavily keratinized cells.
Hair does not grow continuously. A hair
follicle has an active growth period (anagen)
that alternates with a rest period (telogen).The
length of the growth and rest periods ofthe hair
folliclesvary from region to region in the body.
In the scalp, for example, the growth period
lasts for 2 to 4 years and the rest period last for
about 3 months. Meanwhile in the eyebrow,
the growth period is only from 1 to 2 months
while the rest period is 3 to 4 months. During
the rest period of the hair follicle, the hair is
shed, the hair papilla becomes rudimentary, the
follicle becomes smaller and shorter, and the
melanocytes become inactive.
Sheaths of Hair
The internal root sheath and external
root sheath form the outer layers of the hair
follicle.
The internal root sheath surrounds the
initial segment of the hair. It is formed by the
multiplication and differentiation of the most
peripheral cells in the hair matrix. In its deep
segment, the internal root sheath consists
of three concentric layers, but as the cells of
the internal root sheath move. upward, they
degenerate and die such that above the level of
the sebaceous glands, the inner root sheath is
no longer present.
The external root sheath envelops the
internal root sheath. It consists essentially
of the epidermis of the skin that is pulled
downwards by the invaginating hair follicle.
Near the skin surface, the external root sheath
exhibits all the layers of the epidermis of thin
skin. But in the deepest portion of the hair
follicle, it is reduced to a single layer that
corresponds to the stratum germinativum of
the epidermis.
Nerves Associated with Hair
Hair is a tactile organ. It is richly supplied
with free and encapsulated nerve endings
including lanceolate endings-nerve endings
that are in contact with hair bulbs and which
resemble Meissner's corpuscles structurally.
Nail
N ails (nail plates) consist of heavily
keratinized epithelial cells that form protective
covers on the dorsal surfaces of the terminal
ends of the digits. They-grow continuously but
those in the fingers grow faster than those in the
toes. In the same limb, the nail of the middle
digit grows faster than the others.
The nail plate is essentially synonymous
with the stratum corneum of the epidermis.
It rests on a nail bed, which consists of the
stratum spino sum and stratum germinativum
of the underlying epidermis. The cells of the
nail bed do not contribute to nail growth.
Fig. IX-7. Nail. H&E x40.
Growth occurs at the nail matrix, the most
proximal portion of the nail.
The proximal part of the nail plate, the
nail root, is partly hidden in the nail groove.
It extends from the lunula, the white, crescent-
shaped area at the proximal portion to the
exposed nail plate to the nail matrix. In the
nail groove, the nail plate is covered on both
its dorsal and ventral surfaces by layers of the
epidermis. That which covers the dorsal surface
of the nail plate is referred to as the nail fold. At
the lunula, the distal end of the nail fold is highly
keratinized and referred to as eponychium
(natl-cuttcle). The eponychium consists of
the stratum corneum of the overlying skin.
Under the free end of the distal part of a nail is
a thickened accumulation of stratum corneum
called hyponychium.
N ail growth is the function of the cells of
the nail matrix, which corresponds to the cells of
the stratum germinativum of epidermis. These
cells differentiate into a single type of highly
keratinized cells. As they mature, degenerate
and die, instead of getting pushed upward as in
hair, they get pushed forward by younger cells.
In the process, they slide over the nail bed and
form the nail plate.
SKIN AND ITS APPENDAGES ~
Fig. IX-B. Sebaceous Gland and
Other Skin Appendages. The
section shows that the sebaceous
gland is a simple, branched
alveolar gland. Scalp, H&E x400.
Sebaceous Glands
The sebaceous glands are essentially
appendages of hairj hence, they are found in
all areas where hair is present. They are most
numerous on the face, forehead, and scalp
where, during puberty, they may become very
active and give rise to acne and pimples. One
or more sebaceous glands are associated with a
hair and their ducts open into the upper third of
the hair follicle.
Sebaceous glands are generally absent in
hairless skin such as the palms and soles and
the dorsal surfaces of the distal portions of the
digits. However, there are skin areas where hair
is absent but which possess sebaceous gland,
notably the areola and nipple of the mammary
glands, labia minora, clitoris, prepuce, glans
penis, lips, and corners of the mouth.
Sebaceous glands are simple, branched
alveolar glands that are embedded in the
dermis and occasionallyalso in the hypodermis.
They are holocrine glands and it means that
their secretory products consist not only of
the secretion of cells but also of the cells that
produce these secretions.
In routine histologic preparations, the
acinus of a sebaceous gland is seen as consisting
of a single layer of flattened or cuboidal
peripherally-located cells (basal cells) that
ESTEBAN& Cim~Zt\USTEXTBOOK OF HISTOLOGY
rest on a basal lamina and surround a central
mass of larger, paler-staining, fat droplet-
containing cells which are at various stages of
differentiation. The basal cells have a spherical
nucleus, the usual complement of organelles,
and some fat droplets.
The typical sebaceous gland has a short
duct lined by stratified squamous epithelium
that is continuous with the external root
sheath of the hair follicle.
The secretory product of sebaceous
gland is called sebum. It is produced in the
alveoli (acini) of the gland as follows. The
basal cells divide continuously either to renew
their numbers or to differentiate. When the
progenies of the basal cells differentiate,
they become rounded cells that contain
numerous fat droplets in their cytoplasm. As
the differentiating cells mature, they become
larger, paler-staining and accumulate more fat
droplets. At the same time, they get pushed
into the central portion of the acinus by
younger cells. Ultimately, the cells degenerate
and their nuclei become pyknotic. When the
cells die, their cell membranes disintegrate
~I
releasing their fat droplets. The fat droplets
and the remnants of the cells that produce
them constitute sebum. Sebum lubricates hair
and possibly helps maintain the soft texture
of skin, but it hardly plays a role in preventing
fluid loss.
Sweat Glands " numerous on the palms and soles. The only
skin where they are absent are those in the
The skin has two types of sweat glands: margins ofthe lips, concha of the external ear,
eccrine and apocrine. These two types differ nipple, labia minora, and glans and prepuce
structurally and functionally. Eccrine sweat of the penis.
glands are much more numerous than apocrine
The sweat gland is a simple coiled tubular
sweatglands,which iswhythe term sweatgland,
gland. Its duct is lined by stratified cuboidal
unless preceded by a modifier, refers to eccrine
epithelium. The opening of its duct on the
sweat gland.
skin surface is called sweat pore. Its secretory
There are also modified sweat glands that portion, which has a slightly bigger diameter
are present in certain parts of the body, to name than its duct, is embedded deep in the dermis and
a few: the glands of Moll in the eyelids whose the hypodermis. It consists of a pseudostratified
secretions are released into the follicles of the low columnar epithelium that in routine
eyelashasesand which along with the glands of histologic preparations is seen to contain three
Zeis are the usual sitesforstyes, the ceruminous types of cells: light, dark, and myoepithelial
glands of the external auditory canal, which
cells.The light and dark cells are secretory cells.
produce cerumen; and the mammary glands,
The light cells, which rest on a basal lamina,
which secrete milk.
are broad on their luminal portion but tapering
towards the basal lamina. They secrete water
Eccrine Sweat Glands and electrolytes and are responsible for most
Eccrine sweat glands are widely distributed of the watery component of sweat. The dark
in the skin all over the body but are most cells, on the other hand, are broad at their

SKIN AND ITS APPENDAGES 'Iii

bases and narrow at their luminal portions.
They are smaller and fewer than the clear ceils.
They contain dense, glycoprotein-containing
secretory granules in their cytoplasm th'at are
resp0I?-sible for the dark appe.arance of the cells.
As a rule, the bases of the dark cells _donot rest
on the basal lamina-an argument In favor of the
'glandular epitheliumbeing stratified instead
of being pseudostratified. The light and .dark
cells are distinguishable it' special stain{~uch
as PAS are used. The myoepithelial cells, which
are morphologically similar to those present' in
other exocrine glands, are located between the
secretory cells and the basal lamina.
Sweat glands are merocrine glands whose
primary fu~cnon is to help regulate body
remperatur~. They do this by producing a cleat
fluid called sweat that evaporates (;m, and cools,
the skin surface. Sweat also.serves as-a vehicle
for excreting some waste materials and products,
from the body.
Apocrine Sweat Glands
Apocrine sweat glands are found only in the
axilla, around the anus, areola of the breast,
and labia majora. They are also coiledtubiilar
ESTE:B!\f\) & G()f~ZALES'TEXTBOOKOF HISTOLOGY
glands but ~re much larger (secretory portion
= 3 to 5 mm in diameter) than the eccrine
sweat glands (0.4 mm in diameter). They are
embedded deep in the dermis and subcutaneous
tissue. Their ducts, as a rule, open into hair
follicles although a few open directly into the
skin surface.
like in eccrine sweat glands, have associated
myoepithelial cells.
Unlike eccrine sweat' glands, however,
which are active from childhood, apocrine sweat
glands,only become functional at puberty. They
are called apocrine sweat glands because it was
once thought that the secretory cells lose their
apical cytoplasm into their secretion. More
recent findings, however, have established that
they are actually also merocrine"if aniIs.
The apocrine sweat glands playa mino
role in temperature regulation because of their
limited number. Their sec' 'eH811,which is
more viscous than that of eccrine sweat gland,
is sterile and odorless per se, but bacterial
action on the secretion produces an unpleasant'
odor.
Circulatory System'

he circulatory system is the organ initial tributaries' of veins. Arteries bring blood

T system that brings nutrients, oxygen, from the heart to the .capillaries while veins
hormones, and other needed substances bring blood from the capillaries back to the
to the cells of the body from various points of heart. It is across the thin walls of the capillaries
origin. It also movesthe carbon dioxide, and and very small veins that exchange of gases and
waste and secretory products generated by substances between blood, on the one hand, and
the cells to their disposal areas and/or target the tissues, on the other hand, takes place.
•
organs. Additionally, it aids in fighting off The heart and the blood vessels comprise
pathogenic microorganisms by providing and/. , two continuous systems of tubes. One-syste-m,
or transporting the cells and substances needed the pulmonary circulation (pulmonic
for this purpose. circulation), brings blood from the heart to
Two closely related systems make up the lungs and then back to the heart. The other
the circulatory system: the car{liovascular system, the systemic circulation, brings blood
system and the l~mp-hvascular sy-stem.Both from the heart to all the other tissues and organs
systems are composed of hollow channels of the body and then back to the heart.
through which fluid, where substances and
cells are suspended, circulates. In the lymph Heart
vascular system, the circulating fluid is a
milky substance called IrmEli, whereas in the The eart is ahollowmuscular organ,about
cardiovascular system, the circulating fluid is the size of a clenched fist, which is located in
lood. the central mediastinum of the thoracic cavity.
By contracting, it acts as a pump that propels
blood to the arteries of both the systemic and
CARDIOVASCULAR SYSTEM pulmonary circulations.
The cardiovascular system consists of the
lieart and the 1:)Ioodvascular sIstem. Blood
vascular system refers to the Blooil vessels
that form a closed circuit to and from the
heart. There are three types of blood vessels:
capillary, artery, and vein. Capillaries connect
the terminal branches of the arteries to the

Fig. X-1. Organization of the Cardiovascular
System
The chambers on the right side of the
heart are separated from those on the left
by the interatrial sep.tum superiorly, and
by the interventricular septuQI inferiorly.
The chambers on the same side of the heart
communicate with each other through
orifices that are guarded by one-way valves.
The right atrium communicates ';ith the
right ventricle via the rigHt atrioventriculaJj
orifice (tricuspia orifice), which is guarded
by the right a rioventricular va ve Gtricuspia
Ive).The left atrium communicates with the
left ventricle through the left atrioventriculal1
orifice (6icusp.i(l orificej mitral ortfice),
which is guarded by the left atrioventricular
valve (bicuspia valve; mitral valve). The
valves ensure unidirectional flow of blood
through the chambers of the heart.
The chambers of the heart are in
communication with the large blood vessels
of the systemic and pulmonary circulations.
The superior and inferior vena cavae
are continuous with the right atrium: the
pulmonary (pulmonic) trunk with the right
ventricle, the pulmonary veins, two left and
two rights, with the left atrium: and the aorta
with the left ventricle.
ESTEBAN& CCNz/\LES' TEXTBOOK OF HISTOLOGY
In the. ventricles, the orifices of the
pulmonary trunk (pulmonic or if'ic e ,
pulmonary orifice) and the aorta (aorfre
orifice) which are collectively referred to as
~~'Ihilunar ortfices, like the AV orifices, are
guarded by one-way valves, the pulmonic and
aortic valves, respectively. These valves are
collectivelyreferred to as semilunar valves.
Flow of Blood through the Heart
Th~ superior vena cava, inferior vena cava,
and coronary sinus (the vein into which the
cardiac veins drain) bring unoxygenated blood
from the different parts of the body to the right
atrium. Blood then goes into the right ventricle
through the tricuspid orifice. Contraction
of the right ventricle propels blood inside
its cavity into the pulmonary trunk, which
carries the blood to the lungs for oxygenation.
From the lungs, blood is brought back to the
heart, specifically to the left atrium, by the
pulmonary veins. Blood then goes to the left
ventricle by passing through the mitral orifice.
When the left ventricle contracts, the blood
in its cavity is forced into the aorta, whose
branches then distribute the blood to all other
parts of the body.
Pericardium
The heart is envelopedby connective tissue
called pericardium] The pericardium actually
consists of two pouches, namely: fibrous
pericardium and serous pericardium, which
are intimately bound to each other.
The fibrous pericardium, the more
external of the two pouches, lines the
central mediastinum. It is made up of dense
connective tissue. The serous pericardiumj
on the other hand, has two layers because
during development, the heart and the great
vessels,which lie on the internal surface of the
fibrous pericardium, invaginate it from behind.
The more external of the layers of the serous
pericardium adheres to the fibrous pericardium
and is called parietal pericardium while the

Fig. X-2. The Developing Heart and the Layers
of the Pericardium
more internal la f..;r adheres to the heart and
is called visceral pericardium. Between the
parietal and visceral pericardia is a space, the
pericardial cavity, which contains a small
amount offluid (pertcardial flnid'). Pericardial
fluid, which amounts to IS-SO mL in adults,"
reduces friction within the pericardium by
lubricating the luminal surfaces of the visceral
and parietal pericardia, thus allowing the
membranes to easilyglide over each other when
the heart contracts.
The parietal pericardium is made up of
loose connective tissue that is covered on its
free surface by mesothelium. The visceral
pericardium, on the other hand, which is also
covered on its free surface by mesothelium,
is synonymous with the epicardium and
is discussed in the succeeding section. The
mesothelial cells are serous secreting cells.
They are responsible for elaborating pericardial
fluid.
Histologic Layersof the Heart
The wall of the heart has three histologic
layers. From within outwards, these layers
consist ofthe endocardium, myocardium, and
epicardium.
Endocardium
The endocardium isthe thinnest histologic
layer of the heart. It lines all the internal
surfaces of the heart and is continuous with the
innermost layer of the great vessels that enter
and exit the heart.
The endocardium has four histologic
layers. The layer that adjoins the cardiac
lumen is a simple squamous epithelium called
e~do.thelium. Deep in the endothelium
is a very thin layer (sub'ertdotheliallayerj
subendothelium) of loose connective tissue.
External to the subendothelium is a layer of
denser connective tissue, which comprises the
thickest portion of the endocardium. This layer
contains abundant collagen fibers, fibroblasts,
a great number of elastic elements, as well as
smooth muscle cells which are numerous
over the interventricular septum. External to
this layer is another loose connective tissue
layer called sub~endocardium, a misnomer
because the layer is part of the endocardium.
The subendocardium contains many blood
vessels and nerves. It also contains many
of the Purkinje fibers that comprise the
impulse-conducting system of the heart
{see page 141).
..
Fig; X-3. Endocardium and Myocardium. Note
the Purkinje fibers in the myocardium. Heart,
H&E x100.
CIRCULATORY SYSTEM ~
 Fig. X-4. Myocardium. The striated nature of
the cardiac muscle fibers are very evident in this
higher magnification of the same specimen in
Fig. X-3. Heart, H&E x400. .

Fig. X-So Epicardium.

Notice the adipose tissue
that occupies much of the
epicardium. Heart, H&E x100.

Myocardium and brain natriuretic peptide (BN~~-which

participate in cardio-renal homeostasis and
The myocardium, which liesexternal to the
whose target organs are he kidneys, adrenals,
endocardium, is the thickest layer of the heart,
pituitary gland, and brain.
but its thickness varies in the different parts
of the organ. It is thickest in the left ventricle
and is thinnest in the atria. It consists mainly of Epicardium
cardiac muscle fibers (cells) which have been The epicardium, which as previously
described in the chapter on muscle tissue. The . mentioned is synonymous with the !Visceral
myocardial muscle fibers are arranged in sheets Berica~tr.rum, forms the outermost histologic
that wind around the atria and the ventricles in layer of the heart. It consists ofloose connective
a complex, spiraling course. They originate and tissue that is lined on its external surface by
insert in the cardiac skeleton. esoJJlelium.Embedded in the epicardium are
Some cardiac muscle cells in the atria and networks of elastic fibers, blood vessels, and
the interventricular septum have endocrine nerves. In the areas of the heart where bigger
functions. They secrete at least two polypeptide branches ofthe coronary arteries are lodged, the
hormones-atrial natriuretic peptide (ANPy epicardium contains a lot of adipose cells.

ESTES;'",\) & GONZAL.ES' TEXTBOOK OF HISTOLOGY

 Skeleton of the Heart
Cardiac skeleton is the term for the dense
connective tissue that forms the central support
of the heart into which the cardiac muscles and
val~es are attached. It has three components:
a) septum membranaceum, which refers to
the part of the cardiac skeleton that is in the
interventricular septum, b) annuli fibrosi,
which surrounds the AVand semilunar orifices;
and, C) trigona fibrosa, which are between the
semilunar and AV orifices.
Cardiac Valves
The xr and sem:Hunar valves are similar
in structure microscopically. They consist of
reduplicated endocardia and a core of dense
connective tissue that is continuous with the
annuli fibrosis.
Normally, the cardiac valves are avascular.
Fig. X-6. Cardiac Valve. In sections, a cardiac
valve consists simply of a core of dense
connective tissue that is covered on both surfaces
by endocardium. Heert; H&E x200.
Impulse-conducting System of
the Heart
The contraction of the cardiac muscle
fibers is triggered not by neural impulse but
by electrical impulse that is generated and
propagated by a population of modified cardiac
- - -- ---
SA
right
bundle
brafteft
Fig. X-7. Components of the Impulse-
conducting System of the Heart
muscle fibers called 'Purkinje fibers (Purkinje
cells; Purkinje cardiomyocytes) that are non-
contractile.
In H&E preparations, Purkinje fibers are
seen to be larger, paler, and containing more
glycogen in their cytoplasm than the typical
cardiac muscle cells.
The Purkinje fibers interconnect with
each other through a variety of cell-to-cell
attachments including desmosomes, gap
junctions, and fascia adherens to comprise the
impulse-conducting system of the heart.
The impulse-conducting system of the
heart has several components: sinoatrial
node (SA node), three internodal tracts,
atrioventricular node (AV node), AV bundle
of His, two bundle branches and Purkinje
fibers.
The ~A node consists of a dense network
of interwoven Purkinje fibers that are slightly
smaller than those found elsewhere in the heart.
It is about 10 mm in length and 3 mm in width
and is located subepicardially (i.e., underneath
the epicardium) at the boundary of the ri~h~
m-ll..;itn and the sJ!per,iorvena cava. The SA
node is called the ~~dljas=pacemaker because
its Purkinje fibers generate the electrical
impulse (cardiac impulse) that initiates cardiac
contraction.
CIRCULATORY SYSTEM _
 The impulse from the SA node is Anastomoses among the branches of the
propagated to the atrial musculature and coronary arteries exist but they are insufficient
reaches the AV node via three small bundles of in providing alternate routes for blood
Purkinje fibers: the anterior internodal tract circulation in case of obstruction to the major
(of Bachman) j the middle internodal trac (of vessels. Consequently} the coronary arteries are}
Wenckebadi)j and) the posterior internodal .functionally} en(i arte!~~~.
tract (of Thore'i0. Most of the cardiac veins empty into the
The NV nbde is another collection of corona);[ sinus, which in turn opens into the
Purkinje fibers. It is about 6 mm long and 2 to right atrium} but a few drain directly into the
3 mm wide and is located in the myocardium right atrium.
of the posterior lower part of the iqt~ratrial Lymph channels are closely associated with
s~ptllm. the musculature of the heart. They are abundant
From the AV node) the cardiac impulse is in the myocardium} subendocardium, and
propagated into the Ai'J?undle (of His)} which
is the direct continuation of the AV node. The
subepicardium.
The cardiac musculature does not need
--- .
AV bundle (of His) is located in the dense neural stimulation to contract. Nevertheless}
connective tissue of the dgclDum fibrosum.
I
the heart receives efferent nerve fibers from the
It is formed by Purkinje fibers that course iVagusnerve (CN~X)and sy:mpatlletiCaiVisloD 1-
downward towards the ventricles. At the area of the autonomic ner~0US s;}3 ..,tern.The vagal
of the sellti~'ji't"
membranaceurii, the AVbundle fibers inhibit} whereas the sympathetic fibers
bifurcates to form the right and left bundle stimulate heart action.
branches.
The axon terminals of the efferent nerve
The tig:'f~b.lindle b r a n ch runs fibers are not in direct contact with the muscle
downward along the periphery of the septum 'fibers they innervate. They are located a short
membranaceum in the su.}j~naocardium of the distance from the muscle cells. They release
right ventricle. It proceeds to the interventricula their neurotransmitters into the intercellular
septum where it splits into many fine branches space} which then reach the muscle fibers by
that are simply called E.ll1t;lrrhje fibers} which diffusion.
spread through the musculature of the entire
right ventricle.
BLOOD VASCULAR SYSTEM
The l~:ft "b·u.udIe branc is also in the
subendocardium but of the left ventricle. Like Blood vascular system} as already
the right bundle branch} it ramifies and its mentioned} is the collective term for all the
numerous branches (Purkinje fibers) supply the blood vessels in the body} of which there are
myocardium of the left ventricle. , three types: arteries, veins, and capillaries.
As in the heart} the luminal surface of all the
Blood and Lymphatic Vessels, blood vessels is lined by endothelium.
and Nerves of the Heart In capillaries} the endothelium is the only
component of the vessel wall but in arteries and
veins}the vessel wall has other components.

Endothelium
The ,endo__fieHunf serves as a lining
material not merely to facilitate the flow of
blood through 'the blood vessels but also to
I
I
. EST[B/\f~& GO\jZN..[S' TEXTBOOKOF HISTOLOGY
regulate the diffusion of substances and cells to in tendons, nerves, smooth muscles, and serous
and from blood. membranes.
Endothelial cells also secrete some Not all capillaries in any given organ or
substances that are important in the regulation tissue are used Simultaneously. Normally, many
of the cardiovascular system including bloo4 of them are closed and do not contain blood.
do..!t~g factors such as v~!eb(and factor, They open only when the need arises.
~d~Th.elins, p~~cyclip.s, ~iffi£ ...
q~ide, and Capillaries are disposed in different
other substances that mediate the inflammatory planes in most tissues and because many
response. They likewise exhibit phagocytosis, pursue irregular courses, they are seldom
albeit to a limited extent. seen in longitudinal section in thin histologic
Endothelial cells have mitotic capability, specimens.
which they display when a blood vessel is
damaged or needs to increase in caliber. Pericytes (cells of Rouget; mural
cells)
Capillaries
Capillaries are the simplest of the blood
vessels. They have a very thin wall that consists
simply of a Single layer of endothelial cells that
rest on a basallamin:a. Generally, capillaries lie
on a bed of connective tissue.
The Iuminal diameter of capillaries, except Pericytes envelop-but are not part of-
for sinu~oma""capilraries which have bigger the wall of capillaries. They rest on a thin
caliber, is often only 7 to 9 llm. Thus, red blood basal lamina that, in places, fuses with the
cells have to pass through most capillaries in basal lamina of the capillary endothelial cells.
Single file and with difficulty. This "heavy Pericytes probably influence the luminal size
traffic" condition allows time for exchange of of capillaries because they contain-as shown
gases to take place between the red blood cells by immunohistochemistry-actin, myosin,
and the surrounding tissue. tropomyosin and desmin, which make them
In routine histologic preparations, the contractile. Some pericytes are phagocytic while
wall of a capillary, when seen in cross section, some are sources of new endothelial cells.
usually consists of a single endothelial cell
that surrounds the capillary lumen, although Types of Capillaries
occasionally, 2 or 3 cells may be seen. Ga illar}j Electron microscopic studies have
e-n ofli~~lialcells have an ovoid or elongated demonstrated three types of capillaries:
nucleus that bulges into the lumen of the vessel continuous, fenestrated, and sinusoidal.
while their attenuated cytoplasm is clear to
COijJindoqs (type I) capillaries are found in
finely granular.
musclevs,Jungs;CCiifral nervous system, and
All organs are supplied with numerous sJdn. This type of capillary is characterized by
capillaries that form networks that vary in an uninterrupted endothelium where adjoining
number, size, and shape depending on their endothelial cells adhere to each other mainly
location. Capillary networks are abundant by j:~~rdigitatingj other types of cell-to-cell
and dense in the lungs, liver, kidneys, mucous attachments (e.g., desmosomes and zonula
membranes, glands, skeletal muscles, and in occludens) are rare. The cytoplasm of the
the gray matter of the brain. They are sparse endothelial cells contains fine filaments and

CIRCULATORY SYSTEM ..
Fig. X-S. Capillaries in Cross Section. Note that
the wall of the capillaries (at arrows) as seen in
cross section typically consists only of a single
endothelial cell. Cerebrum, H&E x400.
Fig. X-9. Capillaries in Cross and Longitudinal
Sections. This section of the myocardium shows
typical capillaries in cross (cc) and longitudinal
section (lc). Also labeled in the section are some
intercalated discs (ic). Heart, H&E x400.

vesicles that vary in number depending on the The endothelium of sinusoids is formed by a
location of the capillary. mixture of phagocytic and non-phagocytic cells
~estrated (type II) capillaries are found in that rest on a discontinuous basal lamina that is
separated from the parenchyma of the organ by
the mucous membranes of the g~~'toJntestinaF
a very fine network of reticular fibers:
tract, many en.iI~£tineglands) pancreas and
renal glomerulus. In these capillaries) the
cytoplasm of the endothelial cells is very thin Histologic Layers of Arteries and
and is pierced at intervals by "p(Jl1~S"that range Veins
in size from 60 to 80 nm. The pores are bridged
The walls of arteries and veins have three
by diaphragm ..,---exceptin the glomerulus where
histologic layers or coats) namely tunica intima)
the pores have no diaphragms-that consists of
tunica media) and tunica adventitia which are
a unit membrane that is much thinner than the
analogous to the endocardium) myocardium,
cell membrane.
and epicardium) respectively.
S~fiusoidalcapillaries (sinusoidsj are found
in the parenchyma of some organs including Tunica Intima (Tunica Interna)
the liver) sple'en) \rone marrow and certain
endocrine glands (e.g., adrenal cortex). The tunica 'intima consists of an
Unlike type I and type II capillaries) sinusoids endothelmm and a subendothelial layer
have irregular and large cross-sectional outlines. (subendothelium) that is made up of loose
Like fenestrated capillaries) they have true connective tissue that may have occasional
discontinuities in their endothelium that allow smooth muscle cells.
free passage of blood that is why some authors In most arteries) a third histologic layer is
consider them as special types of fenestrated present in the tunica intima. This layer) which
capillaries. In areas) these continuities are consists of the non-fibrillar form of elastin)
covered by basal lamina. In cross section) the lies external to the subendothelial layer and is
circumference of the sinusoidal wall is seen to known as the itY$r~alelastic lamina (internal
be formed not by a single but by several cells. elastic membrane).

ESTE3!\N & Cc)~E.<\LES'TEXTBOOK OF HISTOLOGY

 The endothelial cells of arteries are often
seen in electron micrographs to contain rod-
shaped cytoplasmic inclusions called Weibel-
PJAI.!tdebodies or granules that are about 3.0
~m by 0.1 ~m in siz~:'Weibel-Palade bodies are
storage sites for the von 'Willebrand factors
a large protein that is necessary for normal
coagulation ofblood. The von Willebrand factor
is actually synthesized by endothelial cells of
all blood vessels, but only the endothelial cells
of arteries store it.
Tunica Media
The ~hicainedi~ is mainly made up
of concentrically-arranged smoofli'jltuscle
~~!>ers.Interspersed among the muscle fibers are
some connective tissue elements that include
collagen and elastic fibers. In arteries that have
relatively large calibers; the tunica media has
another layer, the 'exter al elastfc lamina
(external elastic membrane), that is external
to the smooth muscle layer. The external elastic
lamina, like the internal elastic lamina, is made
up of the non-fibrillar form of elastin.
Tunica Adventitia (Tunica Externa)
The tunica adventitia is chiefly made up
of lo1fS'econnectrve tissue where the cells and
fibers are arranged longitudinally.
Arteries
As they move further from the heart,
arteries increase in number because they ramify,
but they progressively decrease in caliber.
Accordingly, arteries are classified into three
types based on caliber, namely, small, medium,
and large. The three types of arteries, however,
differ not only with respect to caliber but also
with respect to other structural features. They
also differ functionally.
Small Arteries (Arterioles)
Small arteries are better nown as
a_!teriole:s.Their diameter ranges from 40 to
400 ~m.
The 11!!tunica intima of the smallest arterioles ,
consists only of endothelium. In larger vessels,
the tunica intima may have an internal e}itgtJo
_lll~!J1;Prane. In either case, there is practically no
subendothelial connective tissue.
The tUJlicamedia of arterioles, on the other
hand, consists of a single layer of smooth muscle
cells in the smaller vessels, but in the larger ones
as many as five complete layers may be present.
Regardless of size, arterioles do not have a well-
defi'ned external elastic membrane.
The h.miIT idventiti~ of arterioles,
meanwhile, is usually thinner than the tunica
media. It is composed ofloose connective tissue
that merges imperceptibly with the surrounding
connective tissue.
Arterioles have relatively thick walls and
narrow lumens that offer considerable resistance
to blood flow. Thus, they are able to deliver
blood to the capillaries under greatly reduced
pressure.
Incidentally, the branch of the smallest
arterioles that connects with a capillary is
referred to as a metlyerioJe or grecapillary.
It is often less than 40 ~m in caliber and the
muscle fibers in its tunica media, which control
the flow of blood into the capillary, do not form
a continuous layer.
Fig. X-10. Histologic Layersof Arteries and
Veins
CIRCULATORY SYSTEM ..
Fig. X-11. Arteriole and Venule. Note that the arteriole has
a thicker wall and better developed tunica media. H&Ex100.
Medium Arteries (Muscular Arteries;
Distributing Arteries)
Medium arteries are often called muscular
arteries because they have a well-developed
ift!!l!C'l. media. They distribute blood to the
different parts of the body, hence, they are also
known as ilj~t,.i8uting artede '. The volume of
blood delivered to the target tissues or organs by
a muscular artery is determined by the state of
contraction of the muscles in its tunica media.
Practically all the named arteries of the
body, except for the elastic ones, are muscular
EST[3M~ & CJNZ!,LES' TEXTBOOKOF HISTOLOGY
Fig. X-12. Muscular Artery. The
section demonstrates the three
histologic layers of a muscular artery:
tunica intima (ti), tunica media (tm)
and tunica adventitia (ta). H&Ex40.
arteries. The biggest muscular arteries are
the Iir.acfiial and .femoral~rteries, while the
smallest are unnamed and are less than O.Smm
in diameter.
The tun!c.a
-=':'. ~~
nffima of muscular arteries
shows three distinct layers: enaothelium,
sul_j!!p:.ll~tlielill1\1,and i~ferna] elastic
~~m,~~ne. The delicate sub endothelium is
made up of elastic and collagenous fibers and a
few fibroblasts. The internal elastic "membrane
is very prominent and in routine his~ologic
sections, it is thrown into folds because of the
postmortem contraction of the muscle cells in
the tunica media.
The urri~a media of muscular arteries is
::J.._ ~
composed mainly oflayers of circularly-arranged
smootIi muscle cells that are embedded in a
small amount of connective tissue. The number
of layers formed by the smooth muscle fibers
depends on the size of the artery, but the layers
can be as many as 40. In muscular arteries, the
ex era_.rl elastic membrane, although much
thinner than the muscle layer, is very prominent
in routine histologic preparations.
Fig. X-13. Muscular Artey. The section, which
was especially stained to exhibit elastic fibers,
shows the internal and external elastic laminae.
Verhoeff's stain x100.

The tunitt:a'adventitia of muscular arteries is
also well-developed. It is sometimes as thick, or
even thicker, than the tunica media. It contains
lymphatic vessels, nerves, an
Vasa Vasorum
'li§;~ vasorurri are small blood vessels that
are present within the wall oflarge blood vessels.
They supply the tissues of the blood vessel wall
that are unable to get oxygen and nutrients hy.
diffusion from the blood that circulates in the
vessel lumen.
:~n.v~lJ'iS,the vasa vasorum may be present
in the tunica media and tunica adventitia. Il_l'
al_:;t~ies,however, they are generally confined
to the adventitia.
Vasa vasorum originate from the arteries is to serve as the grij.p of the cardiovascular
they supply or from neighboring ones.
Large Arteries (Elastic Arteries;
Conducting Arteries)
A large artery, of which the ~rta is the
best example, has a wall that is relatively thin
compared to the caliber of the vessel. Grossly,
in fresh specimens, its wall is yellowish due to
the presence of an abundant amount of elastic
tissue.
The ~n,j.&'l)~ ima of a large artery consists
----
of an en othenun\, a su6enilolR'Jium that
'
is made up of loose connective tissue that has
a sprinkling of smooth muscle cells, and an
Fig. X-:14. Large Artery. Note the numerous
elastic laminae in the tunica adventitia. Verhoeff's
stain, x 40.
Internal elastic membrane that cannot be
appreciated as a distinct structure' because it
abuts on the innermost elastic lamella of the
tunica media.
The 'ttnrlca media is the thickest of the
three histologic layer of a large artery. Its most
notable feature is the presence of concentrically-
arranged sheets of elastin called el~stic
amellae. The elastic lamellae, the reason why
large arteries are better known as ela.§.!jF~rferies"
increase in number with age, 40 to 70 in adults.
In between the elastic lamellae is a variable
amount of connective tissue and smooth muscle
cells, the main cellular component of the layer.
The Sm€)0~n tmlscleccellssynthesize most of the
precursor of the elastin that comprises the elastic
lamellae. The e,~te!lilal~ra~tic membrane, like
the internal elastic membrane, is not distinct in
large arteries because it abuts on the outermost
elastic lamella of the tunica media ..
The tut}ka- adventitia of large arteries is
relatively thin and merges with the surrounding
connective tissue. It contains vasa vasorum.
Large arteries are also called conauctlng
a'Herles because they serve as major conduit
in the transport of blood away from the heart.
Their more important function, however,
system during 8iasto (i.e., when the heart is
resting after a contraction). During s~l'e (i.e.,
contraction of the heart), the elastic arteries
store part of the mechanical energy generated
by the contraction of the heart by stretching
their wall. This stored energy is released during
diastole when the large arteries recoil back to
their original shape and size and in so doing,
propel blood through the arteries.
Variations in Arteries
Some arteries manifest structural
peculiarities that reflect the adaptation of the
vessels to their location or function.
CIRCULATORY SYSTEM '*fI
 In arteries that represent transitions from 'lfilniCaadventitia at the bifurcation of each (i.e.,
elastic to muscular type (e.g., axillary and
common iliac arteries), the tunica media is two or more aortic bodies, on the other hand,
made up of elastic lamellae that are interrupted
by islands of smooth muscle fibers.
When arteries pass suddenly from elastic
to muscular type, as what happens when the
celiac and superior mesenteric arteries
branch off from the aorta, the wall of the initial
segment of the smaller vessel is atypical because
the inner region of the tunica media consists
of concentrically-arranged smooth muscle
fibers while the outer region consists of elastic
lamellae.
The arteries within the skull have thin
walls but their internal elastic membrane is
conspicuous while those within the lungs
resemble veins by having thin walls marked by
reduced amounts of muscle fibers and elastic
tissue.
The tunica media of the arteries of the
lower limbs are better developed than those of
the arteries of the upper limbs while the tunica
media of the umbilical arteries is composed
of two layers of smooth muscle fibers: an
inner layer where the fibers are longitudinally-
arranged and an outer layer-where the fibers are
circularly-arranged.
The coronary arteries have thick walls
that contain more than the usual amount of
elastic tissue.
Sensory Organs Associated with
Arteries
Sensory organs are associated with arteries.
The more important ofthese sensory organs are
the carotid bodies, aortic bodies, and carotid
sinuses. The carotid and aortic bodies are
sensitive to changes in the O2 and CO2 tension
of blood while the carotid sinus is sensitive to
changes in arterial blood pressure.
Carotid and Aortic Bodies
The carotid bodies are two small structures
(3 x 5 mm in size) that are embedded in the
ESTEB/).,N
8.: Cim~ZI\LES'TEXTBOOK OF HISTOLOGY
left and right) common carotid artery. The
are located adjacent to the subclavian arteries
near the aortic arch.
The carotid and aortic bodies have similar
structure and function. They consist of two
types of cells, type I (glomus) and type IJ
(sheath), that are embedded in a connective
tissue stroma that is studded with fenestrated
capillaries.
The gJ,O_P:lUS cells in routine histologic
preparations are pale-staining, round or oval
cells that have few cytoplasmic processes.
Typically, their nucleus is large and they
--;
possess the usual complement of cytoplasmic
organelles. They resemble the chromaffin
cells of the adrenal me dulla in that they have
dense-core cytoplasmic granules that contain
catecholamines.
The ~J.w~athcells,on the other hand, areglial-
like1andthey function merelyassupportive cell .
In routine histologic preparations, compared
to 'glomus cells, their nucleus is more irregular
in shape and they do not contain dense-core
granules in their cytoplasm. In addition, they
have more cytoplasmic processes than glomus
cells. Typically, the cytoplasmic processes of a
sheath cell envelop 4 to 6 glomus cells.
The afferent nerve endings in the carotid,
bodies are branches of the glossopha,rynge'll
neyve'"1 CN IX) while those in the aortic bodies
come from the vagus nerve (CN X).
Carotid Sinus
The ~~rotid sinuse~ refer to the slightly
dilated area in the left and right internal
carotid arteries shortly after they branch off,
from the common carotid artery.
In the carotid sinus, the tunica media of
the internal carotid artery is attenuated and the
tunica
.. adventitia is thic .The tunica adventitia
contains numerous sensory nerve fibers that
form networks. These nerve fibers come
from the glossopharyngeal nerve (CNIX);1
 Fig. X-1S. Medium Vein.
The thickest {ayer of a
medium vein is the tunica
adventitia (TA). The tunica
media (TM) is made up of
several layers of circularly-
arranged smooth muscle
fibers, while the tunica
intima (TI) is rather thin.
H&E x40.

They are baroreceptors that are sensitive to amount of connective tissue into their wall.
stretch and hence, are able to detect changes Functionally, they are like capillaries. They
in blood pressure within the arteries. are sites for exchange of gases and substances
between blood and the surrounding tissue.
Veins The~urfica media (Zanpnly be appreciated
As they travel towards the heart, ',.eillS. in venules that have a luminal diameter of at
typically merge and re-merge to form vessels least SO~m. In large venules, the tunica media
that have progressively bigger caliber and may have several layers of muscle fibers, but the
thicker walls. muscle cells are separated by large amount of
Veins usually accompany arteries but they connective tissue.
are more numerous and their distribution and The tunica adventitia of venules is thick in
courses are more variable. Compared to the relation to the overall thickness of its wall.
arteries that they accompany, veins have bigger
calibers, more irregular lumens, and thinner and
Medium Veins
less elastic walls. Furthermore, the muscular
and elastic elements of the vessel wall are Medium veins include almost all the named
generally more prominent in arteries than in veins and their principal tributaries. Their
their accompanying veins while the veins have diameter ranges from 1 to 9 mm.
more connective tissue elements.
In medium veins, the rJ.!:FifC1'intimconsists
Like arteries, veins are classified into of an endothelium and a thin subendothelial
three types according to their caliber: small layer that is made up of a minimal amount of
(venules), medium (medium-sized), and large connective tissue. The tYD~icamedia, on the
(large-sized) . other hand, is composed of small bundles of
circularly-arranged smooth muscle cells. It is
Small Veins (Venules) thinner than the tunica media of arteries of the
Venules are veins whose diameter is 1.0 mm same caliber. The'tia;pieaadventitia, meanwhile,
or less. which is likewise made up of connective tissue,
The smallest venule s/ are essentially forms the thickest histologic layer of the vein
capillaries that have incorporated a small and may have some vasa vasorum.

CIRCULATORY SYSTEM 'IIfI

Large Veins
The inferior vena cava, superior vena
cava, and pulmonary and portal veins are
examples oflarge veins.
In large veins, the tunica intima is thicker
than, but is otherwise basically identical to,
that found in smaller veins. The tunica media,
on the other hand, is poorly developed and
very few smooth muscle fibers are present,
but some vasa vasorum often exist. The
tunica adventitia, meanwhile, is very thick
and consists of three, albeit poorly distinct,
zones: an internal zone of dense fibroelastic
connective tissue, a middle zone of smooth
muscle fibers that are arranged longitudinally,
and an outer zone consisting of a coarse
network of collagen and elastic fibers. It is
well-supplied with vasa vasorum.
Fig. X-16. Large Vein. Note the very thick tunica
adventitia that contains numerous cardiac muscle
fibers. H&E, x100.
Near their openings into the atria, the
tunica adventitia of the pulmonary veins and
vena cavae may contain some cardiac muscle
fibers.
Venous Valves
Many medium veins are provided with one-
way valves that ensure unidirectional flow of
blood to the heart. These valves are semilunar
ESTU3!\N & GONZ!,LES' TEXTBOOK OF HISTOLOGY
folds that usually come in pairs. They are
essentially inward projections of the tunica
intima and consist of a core of dense connective
tissue that is richly supplied with many elastic
elements and covered on its free surfaces by
endothelium.
Modification in the Organization
of Blood Vessels
The general pattern of organization of
blood vessels dictates that capillaries should
occur between arteries and veins, but this
pattern isbroken with respect to arteriovenous
anastomoses and portal systems.
Arteriovenous Anastomoses (AV
Anastomoses; AV Shunts)
In certain regions of the body (e.g.., in the
skin of the palm, sole,lip, ear and nose, mucous
membranes of the nose and the digestive tract,
erectile tissue, and tongue), there are blood
vessels called arteriovenous anastomoses
(AVanastomoses) that connect small arteries
directly to small veins. When AV anastomoses
are open, much of the blood in the arteries
bypasses the capillary beds and drains directly
into veins.
In the skin of the hands and feet, the AV
anastomoses form a large number of small
histologically recognizable bodies called
g!R1.Pera(Singular: glol1!_us~that are richly
supplied with nerve fibers.
Portal Systems
In ''ffi0iffal system's, an artery or a vein
is interposed between two sets of capillary
beds. The artery or vein that is in between the
capillaries is referred to as a p-DI'Ta:1
vessel.
Portal systems where a vein is interposed
between two capillarybeds existin the digestive
system and the pituitary gland. In the digesti~e
s..~~t~m,a big portal vessel (which is simply
called portal vein~ drains the capillaries of
the gastrointestinal tract then breaks up
into sinusoi s (sinusoidal capillaries) in the
livet. In the pituitary gland, a plexus of
veins, the hypophyseoportal system drains
the capillaries of the median eminence (i.e.,
superior portion of the pituitary gland) then
breaks up into sinusoids in the anterior and
pnstertor lobes of the gland.
Fig. X-17. Portal System. The portal vein, which
carries blood from the capillaries of the digestive
tract to the sinusoids of the liver, is the best
example of a portal vessel in the body.
A portal system where an artery is between
two capillary beds is found in the kidl!~ywhere
the efferent arteriolej which is formed by the
union of the glo~erular capillaries, breaks up
into another set of capillaries in the area around
the kidney tubules. In this portal system, the
efferent arteriole is the portal vessel.
~MPH 'A CULAR SYSTEM
When blood reaches the capillaries, it is
still under pressure, albeit very much reduced.
Consequently, a substantial amount of water
and some plasma proteins leak out from the
cardiovascular system into the interstitial space
(i.e., the space between thecells in the different
tissues of the body) through the walls of the
capillaries. It is the function of another tubular
system, the lymph vascular system, to collect
these escapees from plasma and return them to
the venous side of the circulation.
Fluid from the interstitial space' enters the
lymph vascular system via the smallest tubes
that comprise the system, the lymphatic
capillaries. When interstitial fluid enters the
lymphatic capillaries, it becomes known as
lymph. Lymph is a milky substance which, aside
from water and proteins, contains lymphocytes
and fat droplets (chylomicra).
Lymphatic capillaries unite to form small
lymphatic vessels which, in turn, unite to
form successively larger vessels that ultimately
converge into two large lymphatic ducts that
drain into veins at the base of the neck.
Lymphatic Capillaries
Lymphatic capillaries are present in all
tissues, organs and organ systems of the body
except in cartilage, bone and bone marrow,
teeth, placenta, and the central nervous
system.
Lymphatic capillaries differ from blood
capillaries in several respects. Unlike blood
capillaries that are connected at either end to
Fig. X-1B. Lymphatic Capillaries and Vessels.
As the diagram illustrates, whereas the terminal
branches of arteries are connected to small veins
by blood capillaries, lymphatic vessels start as
blind tubes that merge to form lymphatic vessels.
CIRCULATORY SYSTEM ,..

Fig. X-19. Lymphatic Capillary (Lacteal). In
the small intestine, lymphatic capillaries, called
lacteals, start as blind tubes at the end of the
intestinal villi then unite to form lymphatic
vessels. At unlabeled arrow is-an endothelial cell
that lines the lacteal. Jejunum, H&E x400.
the arteries and veins, lymphatic capillaries
start blindly. Furthermore, compared to blood
capillaries, lymphatic capillaries branch and
anastomose more freely and are more variable
in shape and caliber.
Like blood capillaries, the wall oflymphatic
capillaries consists simply of endothelium, but
the endothelium of lymphatic capillaries is
thinner than that of blood capillaries because
the basal lamina is thin and often incomplete.
Also, they have no associated pericytes.
ESTEBAf-,J& GCNZ!\LES' TEXTBOOK OF HISTOLOGY
Lymphati.c Vessels
. Lymphatic vessels can easily be
distinguished from blood vessels by the large
size of their lumens in relation to the thickness
of their walls.
The wall of small lymphatic vessels is only
slightly thicker than the ~all of lymphatic
capillaries. It is composed of endothelium and
an underlying thin layer of connective tissue
that is mainly made up of collagen and elastic
fibers among which are occasional smooth
muscle cells.
In relatively large lymphatic vessels,
three poorly defined layers corresponding to
the histologic layers of blood vessels may be
appreciated: a tunica intima that consists
of endothelium and an underlying thin
connective tissue layer; a tunica media that is
made up of one or two layers of smooth muscle
cells; and a tunica adventitia that is composed
of bundles of elastic and collagen fibers that
blend with the surrounding connective tissue.
Many lymphatic vessels, like the medium
veins, are provided with valves. These valves, as
in veins, are reduplications of the tunica intima
that encloses an internal framework of dense
connective tissue.
Lymphatic vessels are interrupted along
their course by lymph nodes (see chapter on
lymphoid system).
Fig. X-20. Small Blood and
Lymphatic Vessels. This
photomicrograph demonstrates
the morphological differences
among small veins, arteries, and
capillaries. H&E x400.
Fig. X-21. Medium-sized Lymphatic Vessel. Many medium-sized lymphatic vessels, like this one in
the photomicrograph, are provided with valves. H&E x400.
lymphatic Ducts
The lymphatic vessels all over the body
ultimately drain into the two lymphatic ducts,
namely the right lymphatic and thoracic.
The .ight lymphatic duet is smaller and
shorter than the thoracic duct. All the lymph
that lymphatic vesselscollect from structures on
the right side of the body above the diaphragm
empties into this duct. The right lymphatic
duct, in turn, drains its lymph into the right'
brachiocephalic vein at the junction of the
internal jugular and subclavian veins.
The thoracic duct, on the other hand,
collects the lymph from the parts of the body
that are not served by tributaries of the right
lymphatic duct. It drains its lymph into the
venous system at the junction ofthe left jugtd,ar
ana subclavian veins.
The microscopic structure of the lymphatic
ducts is similar to that of large veins. However,
compared to large veins, the histologic layers of
the lymphatic ducts are less distinct. Also, there
are more muscle fibers in the tunica media of
lymphatic ducts than in large veins.
The tunica adventitia and the outeyegid~
of the tunica media of the thoracic duct are
provided with small blood vessels that are
similar to the vasa vasorum of large blood
vessels.
CIRCULATORY SYSTEM ~

he body is under constant threat
T of invasion
microorganisms
by disease-causing
(i.e., viruses,
bacteria, fungi, and parasites) and other toxic
substances. To ward off these threats, the body
employs several lines of defense.
The first ofthese lines ofdefense consists of
the skin and mucous membranes (mucosae)
that act as physical barriers to the entry of
harmful substances into the body. The skin,
with its epidermis that consists of many layers
of cells tightly bound together by desmosomes,
is impenetrable to most microorganisms
unless injured (i.e., lacerated or abraded).
The mucosae, on the other hand, are easier to
breach physically than the skin, but a variety of
structural and physiologic factors help them in
preventing the entry ofharmful microorganisms
and substances. Examples of these factors are
mucus that protects the mucosal surfaces of
the respiratory, digestive, and genitourinary
tracts: cilia in some epithelial cells that help
move and eliminate toxic substances, and tearsi
and saliva that contain antibacterial substances
including lysozymes that fight off pathogenic
microorganisms.
When its first line of defense fails, the body
unleashes an army consisting of a staggering
number ofcellsthat mounts two types ofdefense
systems (responses, processes): inflammatory
respons and immune response.
t
INFLAMMATORY RESPONSE
(NON ...IMMUNE RESPONSE;
INFLAMMATION)
The inflammatory response is an
immediate but mainly localized process that
starts within minutes of tissue damage or entry
of.a microorganism or a foreign antigen. The
effector cells of this process consist primarily
of phagocytes, mainly neutrophiFs and
macrophages. They are assisted in carrying
out the process by a host of other cells
including eosinophils, basophils, mast cells,
NK cells, and ['-cells. These cells destroy
invading organisms and foreign antigens by
phagocytosis or by releasing substances in their
secretory granules that are toxicto the offending
substances or organisms. They may also release
chemical mediators that attract or facilitate the
entry of phagocytes and other cells that playa
role in the inflammatory process into the area
bf injury or invasion.
The inflammatory response is turned
on by a variety-of ways. Sometimes, invading
microorganisms betray their presence by
producing chemotaxins, chemicals that
attract phagocytes. At other times, the foreign
organisms or substances are detected by the
complement system.
 The complement system is a collection of
more than 20 plasma proteins that are produced
by the liver. These proteins constitute an enzyme
cascade (i.e., the enzymes are sequentially
activated).
The complement system is involved in
both inflammatory and immune responses.
In immune response, it is activated by, and
complements the work of, the antibodies
(immunoglobulins), hence the name. In
inflammation, the system gets spontaneously
activated in the presence of microorganisms
or other foreign substances. The activation of
the complement system during inflammation
has the following effects: production of
chemotaxins; marking off of bacteria with
proteins topsonins) to facilitate phagocytosis;
and releasing cytokines (proteins that act
signaling compounds) and triggering the release
of other mediators 'of inflammation such as
histamine by mast cells and basophils.
Once the inflammatory process _has
been triggered, activated phagocytes (i.e.,
neutrophils and macrophages) congregate in
the injured area within minutes. Thereafter,
they start to engulf and digest the invading
microorganisms or other foreign ele~ents. They
also release cytokines and chemical mediators
that further attract and activate other cells that
can help in eliminating the threat. In addition
to digesting pathogens, macrophages and
other antigen-presenting cells (APCs) which
have limited phagocytic activity process the
antigens they have digested. Then they attac
some parts of the antigens on their surface for
presentation to T-cells, the first step in the
immune response. Because many of the effector
cells of an inflammatory response also initiate
a primary immune response, the immune
response is turned on almost simultaneously
with the inflammatory response. Hence, in case
the threat is not eliminated by the latter, the
former can take over and eradicate it.
The cytokines and other mediators
generated during the inflammatory process
are responsible for the classical local signs
and symptoms of in fl am m at ion (i.e.,
swelling, redness, heat and pain). They also,
contribute to, the systemic signs and symptoms
associated with the pro,cess such as in fever.
Fever occurs when interleukin-I, a cytokine
produced by activated macrophages enters the
bloodstream and reaches the hypothalamus,
the temperature-regulating center in the brain.
The hypothalamus then responds by increasing
body temperature.
SE)
The immune response is a mo,re powerful
body defense system than the inflammatory
response. However, if the invading antigen is
a new on (i.e., it has not previously entered
the body), it takes longer time than the latter
to, mount because unlike inflammation, the
immune resPo,nse is not innate; it must be
developed. Furthermore, it is antigen-specific,
which means it must be developed for every
-a_!!figen (i.e., any substance perceived by
the immune system as foreign to the body
arid which consequently induces an immune
response) that the body encounters for the first
time. Antigens vary in size and composition:
from a few amino, acids to, large and complex
antigenic systems such as those of I>acteria,
viruses, fungi, parasites, and other harmful
metabolites (t~xins~.
The principal effector cells of the immune
resPo,nse are the lymphocytes. They are
aided in carrying out the pro,cess by a variety
of cells. Many of these are effector cells of the
inflammatory resPo,nse.
The b o dy has ab ou t two, tr iIl io n
lymphocytes, accounting for about 1% of body
weight. It is estimated that these lymphocytes
are capable of conferring immunity against as
many as lOll different antigens.
The lymphocytes, as discussed in Chapter
VII, are classified into three types based on
the antigen receptors present on their surfaces.
B-cells have B-cell antigen receptors (BCR),
LYMPHOID TISSUE ~
T-celis carry T-ceII antigen receptors (TCR9,
and NK celts carry natural cytotoxicity
receptors (NCR). Aside from these receptors,
lymphocytes also possess other surface markers
or recep.tors. All T-cells, for example, express the
CD3 molecular complex which is responsible for
signal transmission and mediated via the T-cell
receptor (TCR). They also have either CD4 or
CDS markers. Those with CD4 markets (CD4
'"f!,celIs) can differentiate into helper T-celIs
(T h cells), Those with CD8 markers (CD8+.
T-celIs~can differentiate into cytotoxic T-celIs
(TccelIsj and suppressor T-celIs (Ts celIs).
Incidentally, NK cells are also CD3-positive.
Types of Immune Responses
There are two types of immune responses:
humoral and cell-mediated. The resulting
body defenses to infection or disease that
is conferred by these two forms are called
humoral immunity and cell-mediated
immunity, respectively.
The two types of immune responses are set
in motion as soon as an antigen enters the body.
However, one type predominates, depending on
the nature of the antigen.
Humoral Immunity (Antibody-
mediated immunity; AMI)
Humoral immunity refers to immunity
that is mediated by antibodies. Antibodies
(immunoglobulins) are substances that. are
synthesized by plasma cells. Thus, humoral
immunity is primarily the function of B-celIs
because plasma cells differentiate from B-cells.
Antibodies do not destroy antigens; they
simply bind to the antigens that have triggered
their production. However, by so doing, they
are able to neutralize or help eliminate the
antigens by a variety of ways. Many antigens
(e.g., tetanus and diphtheria toxins)
become harmless when they get attached
to antibodies. Some pathogens (all viruses,
some bacteria and protozoa), on the other
hand, have molecules that they use to enter the
ESTEB/\\j & ClO\iZ!\LES' TEXTBOOK OF HISTOLOGY
cells which they then parasitize. Antibodies
can bind with these molecules to prevent the
pathogen from invading the cells. Antibody
attachment can also immobilize bacteria and
protozoans that swim by means of whip-like
flagelIa. In certain instances, antibodies serve
to pinpoint antigens to phagocytes such as
neutrophils and macrophagest Phagocytes
can then phagocytose them, a process that is
greatly facilitated by the participation of the
complement system.
Humoral immunity is important in
containing many viral and bacterial infections.
It is also the type of immunity that is conferred
by vaccines against many childhood
illnesses.
Cell-mediated Immunity (Clvll)
Cell-mediated immunity is not mediated
by antibodies. It is primarily the function of
T-cells. The effector cells of cell-mediated
immunity are mainly the cytotoxic T-celIs (1'
celI~)~ c
" The Tc cells eliminate antigens that are
inside cell and, therefore, protect cells from ~ ..
antibodies. Their target cells include virus-'
infected cells and cells with intracellular,
bacteria. To do this, they need to destroy the
cells that harbor the microorganisms.. They
likewise target cancer cells'and cells that are
foreign to the body. U'ccells are able to destroy
their target cells by a variety of means. One is
by inducing apoptosis when they release the
proteins in their cytoplasmic granules into the
area where their target cells are.
Cellular immunity is also responsible for
delayed hypersensitivity reaction and tissue and
organ transplant rejection.
Development of an Immune
Response
The entry of a new antigen into the
body elicits a primary immune response.
Subsequent entries of the same antigen elicits a
secondary immune response.
Primary Immune Response do when they attach an antigen on their cell
surface. A co-stimulatory signal, however, can
A primary immune response is designed to
only be provided by an APC to a T-cell if other
eliminate the new antigen and more importantly,
molecules on the surface of a T-cell are able to
produce a population of lymphocytes (i.e.,
bind to corresponding molecules on the APC.
memory B-celfs and memory T-cells that
retain the image of the new antigen in their Macrophages' are effector cells of the
memory. It has a long induction phase time infl arn.uator y response. They are also
(severaldaysto weeks,depending on the antigen APCs because when they engulf antigens by
and the amount of the inoculum) because it is phagocytosis, they do not simply destroy the
a multi-stage process that involves a variety of antigens with their lysosomes, but process the
cells. antigens and attach fragments of the antigens
on their surface forpresentation to naive T-cells.
The primary immune response involves
the following steps: 1) antigen recognition, 2) Dendritic cells (DCs) comprise a family of
lymphocyte activation, and 3) effector phase. APCs that derive their name from the branched
projections (dendrites) they exhibit as mature
Antigen Recognition' cells. They are the most potent of the APCs.
They are also widely distributed not only in all
A new antigen that has penetrated surface lymphoid tissues and organs, but in all other
epithelium invariably and almost immediately tissues in the body. They have limited capacity
encounters an antigen-presenting cell (APC) for phagocytosis, just enough to enable them
because APCs are all over the body. to process antigens and then attach antigeni
APCs are cells that phagocytose, destroy materials on their surfaces for presentation to
and process antigens,and then displayfragments naive CD4+ T-cells.
of the antigen on their surface for presentatio . DCs are classified into two broad
to naive CD4+ T-cells~ They are essential categories: myeloid-related dendritic cell
elements of the immune system because as a (mDC) and lymphoid-related dendritic cell
rule, naive CD4+ T cells (i.e., mature cells that (plasmacytoid dendritic cell; pDC~. The
have been released by the bone marrow and former differentiates from the colony-forming
which have not encountered an antigen yet) unit-monocyte/dendritic cell (CFU-M/
do not differentiate into T h cells unless they DC), fa progenitor cell that it shares with the
are presented an antigen by an APC. Likewise, monocyte. Langerhans cells that are present
naive B-cells do not react to an antigen unless in the epidermis of the skin and other stratified
they are activated by cytokines produced by squamous epithelia are examples of mDC .
helper T-cells (T h cells). Naive CD8+ T-cells Lymphoid-related dendritic cells, on the other
also do not differentiate into Tc cells unless hand, are DCs found in lymphoid tissues. Their
they are activated by APCs or helper T-cells (T h origin has not been definitely established yet.
cells). They probably share a common lineage with
Many cells of the body including lymphocytes (see page 123 for lineage tree of
endothelial cells and NK cells can present bone marrow cells).
antigens to T-cells.However, the "professional" B-cellsoften act asAPCs too. They are able
APCs are the rnacrophages; dendritic cells,' to do so when a new antigen binds a receptot
and B-cells 'because these cells can deliver that is present on their surface. They can also
the two signals (antigen signal and co-s do it when they phagocytose and process an
stimulatory signal) that are needed by naive antigen, and attaches parts of the antigen on
CD4+ T-cells to get activated Other APCs their surface. In any case, despite the presence
deliver only the antigen signal, which all APCs of the antigen on its surface, a naive B-cell does

LYMPHOID TISSUE _
not get activated unless it interacts and receives
the appropriate signal from a T h cell.
Lymphocyte Stimulation •
When naive CD4+ T-cells are presented
an antigen by APCs in the presence of co-
stimulators, they proliferate and differentiate
into any or all following cell types: T~_cells
that synthesize the cytokines needed for cell
med£_ted immunity; Iu.cells that synthesize
the cytokines needed for humoral immunity;
and T h3 cells that elaborate the cytokines that
mediate the inflammatory process.
Effector Phase •
In humoral immunity, when T h2 cells
encounter naive B-cells that have the same
new antigen on their surface, _theyinteract with
the naive B-cells and release the appropriate
cytokines to activate the B-cells. The activated
B-cells then proliferate and their pro~enies
differentiate into plasma cells and memory
B-cells: Plasma cells produce antibodies while
memory B-cells carry the image of the antige
tEat led to their formation and are responsible
for effecting secondary immune responses.
Incidentally, some antigens are T-cell-
independent. They are able to activate naive
B-cells on their own.
In cellular immunity, T h l cells secrete
a variety of cytokines that stimulate CD8i'
T-celIs to proliferate. After 4 to 5 days of
proliferation, the CD8+ T-cells differentiate
into cytotoxic Tvcel ls' (Tc cells), memory
T-cells, and suppressor T-cells (Ts cells). Tc'
cell , as previously mentioned, are effector
cells of cellular immunity. Memory T-cells, like
memory B-cells, carry the image of the antigen
that led to their formation and are responsible
for effecting secondary immune responses.
Suppressor T-cells (Tscells) inhibit or regulate
the activity ofB-cells and other T-cells to ensure
that an immune response does not get out of
hand A primary cellular immune response can
sometimes be brought about without the help
ofT h l cells because there are some antigens that
[sr[f3.:\\,j s GONZk_ES' TEXTBOOK OF HISTOLOGY
can be presented directly by APCs to CD8+
T-cells to enable the latter to proliferate and
differentiate.
Once the offending antigen has ,been
eradicated, all the remaining antigenic-
specific cells that have been generated during
the primary immune response, except for the
memory cells, undergo apoptosi .
Secondary Immune Response
The secondary immune response is elicited
by re-exposure to an antigen that has previously
triggered a primary immune response. It has
a relatively short induction phase (1 to 2 days)
and a more rapid build-up. This is because
when the memory B-cells or memory T-cells
that differentiated during the primary immune
response encounter the specific antig n in
their memory, they are able to immediately
recognize it and rapidly divide and differentiate
into effector cells (i.e., plasma cells and Tccells,
respectively) while at the same time, renewing
their number. Very often, the secondary
immune response is so effective that pathogenic
microorganisms are eliminated before the
person manifests any symptom.
As in the primary immune response, once
the offending antigen in the secondary immune
response has been eradicated, all the remaining
antigenic-specific cells die by apoptosis, except
for the memory cells.
Abnormal Immune Responses
The immune response is vital to survival,
but it sometimes goes awry.
Overwhelming reaction to an antigen
causes allergic reactions that are occasionally
fatal, as in cases of anaphylactic shock
following bee stings. Sometimes, the immune
defense system fails to distinguish between self
and non-self antigens. Then the effector cells
attack the body's own tissues and cells, as in the
case of a number of autoimmune diseases.

Fig. XI-1. Loose Diffuse Lymphoid Tissue. The
cells in the tissue are mostly lymphocytes (at
arrows) that are relatively few and far between.
Trachea, H&E x400.
COMPONENT,S OF THE
LYMPHOID SYSTEM
The various tissues (lymphoid tissues)
and organs (lymphoid organs) concerned with
the immune response comprise the lymphoid'
system.
Lymphoid tissue refers to tissue where the
parenchyma consists mainly of lymphocytes
while lymphoid organs refer to organs that are
primarily made up of lymphoid tissue. The
stroma of lymphoid tissues and organs, except
for the thymus, is formed mainly by reticular
fibers and reticular cells.
As mentioned, in the previous .chapter,
the lymphoid tissues and organs are classified
into central (primary) lymphoid organs and
peripheral (secondary) lymphoid tissues and
organs.
The central lymphoid organs are the bone
marrow (described in the previous chapter) and
the thymus. In the central lymphoid organs, the
stem cells (i.e., B stem cells and T stem cells)
transform into mature and immunocompetent,
albeit naive, lymphocytes. They are then
released via blood or lymph into the peripheral
lymphoid tissues and organs.
The peripheral lymphoid tissues and
organs consist of the lymph nodes, spleen
and the mucosa-associated lymphoid tissue
(MALT).MALT includes the tonsils and the
other non-encapsulated lymphoid tissues
in the gastrointestinal, respiratory, and
genitourinary tracts. They are responsible for
initiating immune responses and generating
lymphocytes that participate in these responses.
lymphoid Tissue
Lymphoid tissue exists in the body in the
form of either diffuse lymphoid tissue or
nodular lymphoid tissue.
Diffuse Lymphoid Tissue
Diffuse lymphoid tissue refers to
lymphoid tissue where the lymphocytes
are evenly dispersed. It is a part not only of
lymphoid organs, but also of most connective
tissues in the body. It is especially prominent
in the lamina propria and submucosa
of the gastrointestinal, respiratory, and
genitourinary tracts.
Most ofthe cells in diffuse lymphoid tissue
are T-cells,although there is a small population
of B-cells, plasma cells, and dendritic cells.
Also, some fixed macrophages are associated
Fig. XI-2. Dense Diffuse Lymphoid Tissue. The
numerous lymphocytes (black dots) in the tissue
are packed closely together. Ileum, H&E x100.
LYMPHOID TISSUE ~
 Fig. XI-4. Secondary Lymphoid Nodule.
Fig. XI-3. Solitary Primary Nodule. Note the The nodule has a pale germinal center (GC)
even distribution of the lymphocytes in the surrounded by a dark peripheral region called
nodule. Colon, H&E x400. corolla (C). Lymphoid nodules are usually
embedded in loose or dense lymphoid tissue
(DL). Lymph Node, H&E x400.

-
with the reticular fibers and reticular cells,
and a small population of free macrophages
likewise exists among the lymphocytes.
Diffuse lymphoid tissue where the
lymphocytes are relatively few and far apart
is called loose lymphoid tissue. Diffuse
lymphoid tissue where the lymphocytes are
numerous and close to each other is' called
dense lymphoid tissue.

Nodular Lymphoid Tissue (Lymphoid

nodule; Lymphoid follicle; Lymphatic
follicle)
Fig. XI-S. Aggregates of Lymphoid Nodules
Nodular, lymphoid tissue refers. to (Peyer's Patches). In some areas of the body,
lymphoid _,tissue~,where
clustered lymphocytes lymph nodules(n)embedded in dense lymphoid
tissue (dl) clustertogether. Ileum, H&E x40.
form' discrete ovoid masses or lumps called
lymphoid no~ules.
submucosa of the gastrointestinal, respiratory,
In lymphoid nodules, the lymphocytes
and genitourinary tracts. In the lamina propria
are mainly B:cells. However, a scattering of
and submucosa, lymphoid nodules can occur
T-cells, dendritic cells, and macrophages are
also present. ' . singly (solitary nodule) or in aggregates, as
in the ileum, where they are called Peyer's
Lymphoid nodules begin to-appear only
patches.
at birth. They are usually interspersed in areas
of dense, diffuse lymphoid tissue. They are Under the light microscope, two types of
found in some lymphoid organs (i.e.,spleen and lymphoid nodules are distinguishable: primary
lymph nodes) and in the lamina propria and and secondary.

ESTEBAN & CONZl\LES' TEXTBOOK OF HISTOLOGY

 A secondary nodule is where the
lymphocytes react to an antigen. It has two
distinct regions: a pale central area called the
germinal center, or reaction center, and
a darker peripheral region called corona.
The germinal center is populated mainly
by activated B-cells. These cells undergo
proliferation and functional differentiation
following exposure to an antigen. The germinal
center is paler than the corona because the
lymphocytes it contains are actively-dividing
and young. Therefore, they are larger and have
more abundant cytoplasm and finer chromatin
granules than the smaller, more mature
lymphocytes that form the corona.
In the secondary nodule, the older
lymphocytes are pushed to the periphery as new
cells are produced in the germinal center.
A primary nod,ule; on the other hand
is a nodule where the lymphocytes are idle
or resting. Consequently, it does not have a
germinal center. The lymphocytes it contains
are all small lymphocytes that are evenly
distributed throughout the nodule. " almost exclusivelyT lymphocytes.In children,
Thymus
The thymus is derived from the third
branchial (pharyngeal) pouch. During its
development, it moves caudally and ultimately
ends up in the superior mediastinum, just above
the heart.
At birth, the thymus weighs 10 to 35 g.
In childhood, its size increases in a manner
proportional to the increase in body weight.
It attains its maximum weight (30 to 50 g) at
puberty. Then it gradually involutes, so that
among the elderly, it is reduced to a mass of
connective tissue with a small amount of
parenchyma. Nevertheless, the parenchymal
tissue continues to function. The thymus also
involutes in individuals who are subjected
to severe stress, serious illness, or ionizing
radiation. However, this type of involution is
followed, at least in experimental animals, by
an almost complete regeneration of the organ.
The thymus is the central lymphoid tissue
that is tasked to transform T stem cells into
mature, immunologically competent, and
self-tolerant, albeit naive, T-cells. It also needs
to produce cytokines that regulate T-cell
proliferation, maturation, and function in the
thymus and other lymphoid tissues and organs.
The lymphocytes that populate the thymus are
however, it is normal to find a small population
ofB-cells and even an occasional lymph nodule
in the organ.
While in the thymus, the cells of the T-cell
lineage are often referred to as thymocytes.
Grossly, the adult thymus is composed of
two pyramidal lobes fused together.
Fig. XI-6. Thymus. The capsule (c)
of the organ sends connective tissue
trabeculae (t) into the organ that
divides it into lobules. The parenchyma
of the thymus consists of a peripheral
cortex (co) and an inner medulla (me).
The trabeculae do not extend deep
enough into the medulla so the medulla
of adjacent lobules interconnect with
each other. H&E xB.
LYMPHOID TISSUE
Histologic Organization of the
Thymus
Each of the two lobes of the thymus
is enclosed by a capsule, a dense irregular
connective tissue. The trabeculae from the
capsule divide the lobes, albeit incompletely,
into lobules ofunequal sizeswhile thin strands
of connective tissue from the trabeculae form
secondary septa within the gland.
In routine LM preparation, a thymic lobule
exhibits a peripheral, darker-staining region
(cortex) that surrounds a central, lighter-
staining region (medulla). The cortex is darker
than the medulla because its cells are more
numerous and closely packed. The trabeculae
separating the lobules do not extend deep
enough to separate the medulla of adjacent
lobules; thus, the medulla o[ adjacent lobules
interconnect.
The stroma of the thymic lobules is unlike
that of other lymphoid tissues and organs
because it is not made up of reticular cells and
reticular fibers. Rather, it consists mainly of
stellate cells called epitheloid cells (epithelial
reticular cells) and a few connective tissue
elements.The epitheloid cellsresemble reticular
cells, but they arise from endoderm and do not
produce reticular fibers. They have cytoplasmic
processes that are attached to those of other
epitheloid cells by desmosomes. Six types of
epitheloid cells are distinguishable in electron
micrographs.
The epitheloid cellsare likewiseresponsible
for producing most of the thymic cytokines,
although thymocytes also produce cytokines.
In the cortex, some epitheloid cells (nurse
cells) envelop multiple young lymphocytes.
The nurse cells promote proliferation and
differentiation oflymphocytes.
New T stem cells that arrive from myeloid
tissue (postnatally synonymous with bone
marrow) settle in the peripheral part of the
cortex (outer cortex) where they transform
into lymphoblasts and proliferate. As they
mature, they move towards the medulla while
ESTEB;\N& CCNZ/\LES' TEXTBOOK OF HISTOLOGY
progressively becoming smaller such that
when they reach the medulla, they are mostly
small lymphocytes. During their stay in the
cortex, the T-cells acquire their receptors.
Those that are unable to do so die by apoptosis
(programmed cell death) and are devoured by
macrophages.
In the medulla, the small lymphocytes as
well as the macrophages are not as numerous
as in the cortex. Among the parenchymal cells
in the medulla are APCs known as thymic
interdigitating dendritic cells. These cells
help in preventing autoimmunity by presenting
self-antigens to enable the T-cells to recognize
"self"antigens. T-cellsthat react to the antigens
are induced to undergo apoptosis, and their
remnants are phagocytosed by macrophages.
Most of the lymphocytes generated in the
cortex do not survive the rigorous processing
and screening that they go through. Only
about 10%to 30% leave the thymus as mature,
immunocompetent, and "self-tolerant," albeit
naive, T-cells.
Mature T-cells join the "recirculating
pool" of lymphocytes by entering the blood or
lymphatic vessels in the thymic medulla. From
the thymus, some T-cells preferentially go to
the spleen where they stay for a while, but these
T-cellsultimately join the "recirculating pool"
In routine LM preparations, the most
distinctive feature of the thymic medulla is
the presence of thymic corpuscles (Hassall's
bodies or corpuscles). These structures
are composed of a core of hyaline material
surrounded by layers of flattened epitheloid
cells. The thymic corpuscles which can grow to
100 ~m in diameter increase with age.
Blood Vesselsof the Thymus
The thymus has a rich blood supply that
comes from branches of the internal thoracic,
anterior intercostal, and inferior thyroid
arteries. In the thymus, the arteries ramify
and their branches travel within the trabeculae
and septa. At the corticomedullary boundary,
 continuous endothelium) and their thick
basal lamina; 2) epitheloid cells and their
basal lamina which form a continuous sheet
with the help of desmosomes that tightly
bind adjacent epitheloid cells to each other;
and 3) perivascular space that separates the
endothelium from the layer of epitheloid cells
and contains macrophages and fluid.
The medulla of the thymus lacks a blood-
thymus barrier. To protect the developing
T-ceils, macrophages simply phagocytose the
antigens that are able to enter the area.

Fig. XI-7. Hassal's Corpuscle. A distinctive

Lymphatic Vesselsof the Thymus
feature of the thymus is the Hassal's corpuscle (H)
found in the medulla. The corpuscle consists of a The thymus has no afferent lymphatic
core of hyaline material surrounded byepitheloid vessels. Lymph that is formed within the organ
cell (ec). Thymus, H&E x400.
is collected by efferent lymphatic capillaries
that start as blind tubes. The lymphatic
the arteries give off capillaries that supply the
cortex while the maih branches continue into capillaries unite to form progressively bigger
the medulla where they break up into capillaries. vessels that follow the course of the veins and
exit the thymus through the capsule.
In the cortex, the capillaries follow
tortuous and recurrent routes and anastomose
Lymph Node
extensively. They are drained by postcapillary .
venules which proceed to the medulla where A lymph node is a bean-shaped organ. It is
they unite with the postcapillary venules that simply an encapsulated collection of lymphoid
drain the medullary capillaries to form bigger tissue. There are SOOto 600 lymph nodes in the
venules. Itis in the postcapillaryvenules of the body. They are interposed along the course of
medulla that the lymphocytes join circulating lymphatic vessels. They vary in size (pinhead to
blood by migrating through the blood vessel 3 ern in diameter) depending on their location
walls. and the state of activity of the lymphocytes
The venules unite to form bigger vessels within the organ.
that traverse the connective tissue septa and Lymph nodes are important sites for
trabeculae and exit through the capsule. producing the lymphocytes needed in mounting
an immune response. In these organs, activated
Blood-Thymus Barrier B- and T-cells can proliferate and differentiate
In the cortex of the thymus, the endothelial into various functional types.
cells that line the capillaries form a close The lymph nodes are also highly efficient
relationship with the epitheloid cells to comprise filters for particulate matter and microorganisms
what is referred to as the blood-thymusbarrier. that are present in the lymph. The filtering is
This barrier prevents antigens that are carried performed primarily by macrophages that are
by the capillaries from getting into contact with associated with the lymphatic vessels in the
the developing T-cells. organs. Macrophages phagocytose particulate
The blood-thymus barrier consists matter and microorganisms from lymph and
of the 1) endothelial cells (that form a together with other APCs that also abound in

LYMPHOID TISSUE \B
 hilus

Fig. XI-B. Lymph Node. H&E x40.

the organ. They likewise present antigens to Histologic Organization of the

lymphocytes. Lymph Node
Lymph nodes are strategically situated ·in The lymph node is also enclosed by
various parts of the body. They are present in a capsule, a dense irregular connective
the popliteal, inguinal, and axillary regions; tissue. The capsule gives off the trabeculae
along the sides ofthe neck; along the abdominal that incompletely subdivide the organ into
vessels; in the mesentery; and in many other compartments, whose stroma is formed by
sites. The pattern of their distribution ensures reticular tissue.
that all the lymph throughout the body passes The parenchyma of the lymph node is
through several lymph nodes before reaching divided into a peripheral-located cortex that
the major lymphatic vessels (i.e., thoracic duct enclosesa centrally-located medulla. It consists
and right lymphatic duct). mainly of lymphocytes, but macrophages,
plasma cells, and Des also abound. The
Lymph nodes may become swollen and boundary between the cortex and medulla is
tender when an infection is present in their
"area ofjurisdiction." For example, the inguinal
lymph nodes enlarge and become painful when
an infection develops in the lower extremity.
Grossly,a lymph node exhibits an indented
area called hilus. It is at this area where efferent
lymphatic vessels leave,and blood vesselsenter
and exit the organ. Afferent lymphatic vessels
enter the lymph node on its convexsurface.

Fig. XI-9. Lymph Node Cortex. The cortex of the

lymph node which lies immediately deep in the
capsule (c) can be divided into an outer cortex
(oc)-where lymphoid nodules are present-and
an inner cortex (ic). H&E x100.

ESTEB;.\N & GONZALES' TEXTBOOK OF HISTOLOGY

indistinct. It is arbitrarily delineated by the tips
of the trabeculae.
In the cortex, the outer region (outer
cortex) is mostly occupied by lymphoid
nodules that are embedded in dense lymphoid
tissue. On the other hand, the inner region
(inner' cortex) has no lymphoid nodules and
consists simply of dense lymphoid tissue.
Fig. XI-10. Lymph Node Medulla. This region,
of the lymph node consists of lymphatic vessels
called medullary sinuses. These are separated
from each other by strands of dense lymphoid
tissue called medullary cords. H&E ~·100.
The Des in B-cell-rich areas (outer
cortex) are called follicular dendritic cells.
They have large, pale nuclei and indistinct
borders. Their numerous processes (dendrites)
are joined with those of other dendritic cells
by desmosomes. The Des in Tvcel l-r ich
areas (deep cortex), on the other hand, are
called interdigitating dendritic cells. They
exhibit polymorphic nuclei and processes that
interdigitate.
The medulla is paler-staining than the
cortex. It is made up of dense lymphoid tissue
arranged to form strands called medullary
cords,in between which are medullary sinuses.
Although made up of dense lymphoid tissue, the
cells in the medullary cords are mostly B-cells
and plasma cells,but they rarely form lymphoid
nodules.
Lymphatic Vesselsof the Lymph
Node
As they approach the lymph node,
afferent lymphatic vessels ramify and give
rise to smaller branches that enter the node
on its convex surface. The afferent lymphatic
vessels are provided with one-way valves
that prevent the backflow of lymph. They
penetrate the capsule of the lymph node and
become continuous with the subcapsular
sinuses that lie immediately underneath the
capsule. The subcapsular sinuses, on the other
hand, continue into the trabecular (cortical)
sinuses that travel inward along the trabeculae.
The trabecular sinuses, in turn, carryon into
the bigger medullary sinuses in between the
medullary cords. The medullary sinuses unite as
they approach the hilus to form several efferent
lymphatic vessels. These vessels penetrate the
capsule and leave the lymph node at the hilus.
The sinuses are irregularly-shaped
lymphatic vessels. Their walls which permit all
the constituents oflymph to pass freely consist
simply of an endothelium. The endothelial
cells also serve as APes and thus, they have
limited capacity to engulf particulate matter.
Associated with the endothelium of the sinuses
are numerous macrophages.
The efferent lymphatic vessels are fewer
but bigger than the afferent lymphatic vessels.
Like the afferent lymphatic vessels, they are also
provided with one-way valves which ensure that
the lymph that leaves the lymph node does not
flow back in.
Thus, lymph enters a lymph node via the
afferent lymphatic vessels. Itthen passes through
the sinuses for filtering (via phagocytosis) by the
macrophages, endothelial cells, and other APes
before draining out of the organ via the efferent
lymphatic vessels.
Blood Vesselsof the Lymph Node
The artery that supplies a lymph node
enters the organ at the hilus where it breaks up
into branches that travel within the trabeculae
LYMPHOID TISSUE \B
Fig. XI-11. Lymphatic Vessels of the I ,$I~'us
Lymph Node. Afferent lymphatic
vessels enter the lymph node by
penetrating the capsule on the
convex surface of the organ. The
afferent lymphatic vessels drain into
subcapsular sinuses which, in turn, .}:..~~. !i~.)
drain into the trabecular sinuses that
convey the lymph to the medullary
sinuses. It is then collected by efferent
lymphatic vessels that exit the organ
\:r·.
aff~rent -
Iym-phatic vessel
at the hilus. Lymph Node, H&E x100.
sinus
into the medullary cords. Here) they give off
capillaries to supply the medulla. The main
arterial branches then proceed to the cortex to
form capillary plexuses around the lymphoid
nodules.
The capillaries supplying the cortex drain
into venules (post-capillary venules) that
travel inwards into the medulla to unite with
the medullary venules. Then they form larger
vessels that travel with the arteries in the
trabeculae. The veins ultimately leave the node
at the hilus.
Lymphocytes enter the lymph node
primarily from blood by passing through the
endothelial lining of the blood vessels in T-cell-
rich areas. This ensures interaction between B-
and T-cells.
The lymphocytes leave the lymph node to
join the recirculating pool primarily via the
efferent lymphatic vessels.
Spleen
The spleen is the largest (7 em x 12 em)
lymphoid organ in the body. It is a soft and
freely movable structure that is located on the
left upper quadrant of the abdominal cavity
posterior to the upper part of the stomach.
Its medial surface presents a notch) the hilus,
where blood vessels (splenic vessels) enter and
leavethe organ.
ESTE8;\N & GONZj\LES' TEXTBOOK OF HISTOLOGY
.\ \
fir'·
Like the lymph nodes, the spleen is a very
important component of the body's immune
defense system. In this organ)the lymphocytes)
when activated by blood-borne antigens)
proliferate and differentiate into different
functional types.
The spleen also filters blood. It has an
enormous number of macrophages that destroy
foreign substances, microorganisms, and
abnormal cells present in the blood. It also
removes and destroys damaged and old red
blood cells and platelets from circulating blood
while recycling the iron that is contained in the
RBCs. It also acts as storage area for blood.
The spleen is not a vital organ. In adults)
it can be surgically removed (splenectomy)
without any apparent adverse effect. In very
young children) however) splenectomy has
been associated with increased susceptibility
to certain infections and diseases such as
pneumonia) meningitis) septicemia) and
malaria.
Histologic Organization of the
Spleen
Like all lymphoid organs) the spleen is
encased by a capsule made up of dense irregular
connective tissue. Unlike other lymphoid
organs) however) the splenic capsule contains
a significant number of smooth muscle cells
 Fig. XI-13. Spleen. This higher magnification
photomicrograph of the spleen shows the capsule
(c), trabeculae (t), white pulp (wp), red pulp (rp),
Fig. XI-12. Spleen. The spleen is encased by central arteries (ca), and a follicular artery (fa).
a capsule (c), a connective tissue, that sends H&E x100.
trabeculae (t) into the substance of the organ
to divide it into incomplete compartments. The White Pulp
parenchyma of the spleen is referred to as splenic
pulp. It consists of islands of lymphoid tissue
The white pulp consists of lymphoid
collectively called white pulp (wp) surrounded by nodules that are embedded in dense lymphoid
red pulp (rp). The pulps are made up of sinusoids tissue.
that are separated by reticular tissue. H&E x40.
The dense lymphoid tissue ofthe white pulp
" is organized to form sleeves around the arteries
and elastic fibers. Externally, the capsule is
of the spleen from the time these vesselsbranch
completely enveloped by peritoneum, thus,
the spleen is lined on its externalsurface by offfrom the trabecular arteries to shortly before
mesothelium. they break up into capillaries. The lymphoid
nodules, on the other hand, are interspersed
Connective tissue trabeculae from the along the course of these arterial sleeves.
capsule ramify and form septa that divide the
substance of the spleen, albeit incompletely, In short, the white pulp comprises the
into compartments. The stroma of these lymphocyte population of the spleen which,
compartments, as in the lymph node, consists curiously, is uniquely associated with the blood
of reticular tissue. . vessels.
Aside from lymphocytes, the white pulp
Parenchyma of the Spleen (Splenic also contains numerous macrophages and
Pulp) dendritic cells (splenic dendritic cells).
Gross examination of the cut surface of
Red Pulp
a fresh specimen of the spleen shows that the
parenchyma of the spleen (splenic pulp) is The red pulp-which forms the greater
composed ofa reddish brown substance in which part of the splenic parenchyma-consists of
small masses or islands of ovoid grayish-white numerous large, blood-filled sinusoids (splenic
structures are scattered. The former, which sinusoids) that are separated by strands of
forms the bulk of the parenchyma, constitutes reticular tissue known as splenic cords (of
the red pulp while the latter comprise the white Billroth). The blood in the sinusoids accounts
pulp. for the red color of the pulp.

LYMPHOID TISSUE ~

Fig. XI-1S. Blood
Vessels of the Spleen
Blood Vesselsof the Spleen
The spleen is supplied by the splenic
artery, the biggest branch of the celiac artery.
As it approaches the spleen, the splenic artery
divides into several branches that enter the
spleen at its hilus. These branches ramify to give
off trabecular arteries that course within the
connective tissue trabeculae.
A trabecular artery gives off branches
(central arteries, arteries of the white pulp)
that leave the trabecula to enter the white pulp.
The central artery is a small muscular artery.
ESTEBAN& (10NZI\LLS' TEXTBOOK OF HISTOLOGY
Fig. XI-14. Spleen, Red
Pulp. T!;,ered pulp is made
up of sinusoidal capillaries
called splenic sinusoids
(s) that are separated by
strands of reticular tissue
referred to as splenic cords
of Billroth (c). H&E x400.
penicillary artery draining
reticular -.
tiber network
However, it is atypical because its tunica
adventitia is formed not by ordinary connective
tissue as in ordinary muscular arteries, but by
dense lymphoid tissue, known as periarterial
lymphoid sheath (PALS). As in all dense
lymphoid tissues, PALS consists mostly of
T-cells.
Despite its name, the central artery is
usually eccentrically located in the white pulp.
It courses towards the red pulp, but along the
way, it gives off lateral branches (follicular
arteries) that supply the lymphoid nodules
and dense lymphoid tissue of the white pulp.
PALS likewise envelops all the branches of the
central artery in the white pulp.
At the boundary of the white and red
pulps, the central artery terminates by giving
off branches (penicillar arteries, penicilli,
arteries of the red pulp) to the red pulp. The
penicillar artery is an arteriole, but atypical
because its endothelial cells are cuboidal. It
terminates by giving off two to three sheathed
arteries.
The sheathed artery (ellipsoid) is short
and narrow. It is so-called because it is usually,
but not always, enveloped by concentric layers
of reticular ·cells (sheath of Schweigger-
Seidel). The sheathed artery is surrounded
by macr 'lages, thus, filtering of blood in the
spleen starts here.
Blood from the sheathed arteries drains
into the splenic sinusoids by what is termed as
"open circulation." The sheathed arteries end
blindly and the blood they carry flows out into
the interstitial spaces (among the reticular cells
of the splenic cords) then percolates into the
sinusoids.
The splenic sinusoids interconnect
extensivelywith each other. These blood vessels
have large (up to 40 urn) regular lumens and
very thin walls that do not contain smooth
muscle fibers. Their endothelium is atypical.
The endothelial cells are fusiform and are
capable oflimited phagocytosis. Their ends are
in contact with one another and occasionally
form junctional complexes. However, there are
gaps between their lateral surfaces and they
rest on a discontinuous basal lamina. There
are numerous macrophages (perisinusoidal
macrophages) just external to the endothelial
cells. They are mainly responsible for the
filtering ofblood that occurs in the spleen. The
perisinusoidal macrophages have processes that
extend into the lumen of the sinusoids, which
help the macrophages phagocytose particulate
materials present in the sinusoids.
From the sinusoids, blood flows into
collecting veins in the red pulp. These in turn
drain into trabecular veins which unite to form
the splenic vein that leaves the spleen at the
hilus.
Lymph Vesselsof the Spleen'
The spleen,like the thymus, has no afferent
lymphatic vessels but has efferent vessels that
start as blind capillaries. The -1.pillariesunite
to form bigger vessels that follow the course of
the veins.
Mucosa-associated Lymphoid
Tissue (MALT)
Mucosa-associated lymphoid tissue
(MALT) refers to the enormous amount of
lymphoid tissue that exists in the mucosa
and submucosa of the gastrointestinal,
respiratory, and genitourinary tracts. MALT
associatedwith the gastrointestinal tract is often
referred to as gut-associated lymphoid tissue
(GALT) while that associated with the trachea
and bronchi is sometimes called bronchus-
associated lymphoid tissue (BALT).
In most mucosal surfaces, MALT consists
simply ofloose or dense diffuse lymphoid tissue
that may contain an occasional solitary lymph
nodule. In certain areas of the body, however,
MALT form larger collections of lymphoid
tissue that feature aggregates of lymphoid
nodules. Notable areas-where aggregates of
lymphoid nodules embedded in dense lymphoid
tissue exist-are the colon, vermiform
appendix, ileum (where the aggregates are
referred to as Peyer's patches), and the entrance
to the respiratory and digestive tracts where
the lymphoid tissue aggregates are called
tonsils.
The larger aggregates of MALT are not,
or only partially, enveloped by a capsule. They
consist ofT-cell-rich areas (diffuse lymphoid
tissue) and B-cell-rich areas (lymphoid
nodules). They also contain numerous APes.
MALTs are also important sites for
generating the lymphocytes that are needed in
mounting an immune response.
LYMPHOID TISSUE ~
 Fig. XI-16. Palatine Tonsil. The
tonsil is covered on its surface by a
nonkeratinized stratified squamous
epithelium (e) that invaginates in
places to form tonsillar crypts (tc).
Deep in the epithelium is a connective
tissue (ct) that forms an incomplete
capsule. The parenchyma consists of
lymphoid nodules (n) embedded in
dense lymphoid tissue (dl). H&E x40.

--. .

Tonsils The lingual tonsils consist of several

discrete masses located in the dorsum of the
The tonsils form a ring (Waldeyer's
posterior tongue. Each has a diameter of
ring) underneath the epithelium around the
about 2 to 3 mm. Their superficial surface,
entrances to the respiratory and digestive
like that of the pharyngeal tonsils, is covered
passages. They consist of paired palatine
with nonkeratinized stratified squamous
tonsils, lingual tonsils and tubal tonsils, and
epithelium. This epithelium may have a single
an unpaired pharyngeal tonsil.
broad deep crypt into which ducts of mucus-
Most tonsils are partially encapsulated by
dense irregular connective tissue capsule.
In the lymphoid nodules of the tonsils, the
lymphocytes are usually more abundant on the
epithelial side of the nodules. They were also
demonstrated to be capable of passing through
the epithelium to the surface.
The palatine tonsils are located in
the lateral aspect, one on each side, of the
oropharyxn. Each is an ovoid body that is
lodged in between the palatoglossal fold
anteriorly and the palatopharyngeal fold
posteriorly. The superficial surface of the
palatine tonsils is covered by nonkeratinized
stratified squamous epithelium that forms
deep, sometimes branching, invaginations
called tonsilar crypts that give the tonsils a Fig. XI-17. Lingual Tonsil. The lingual tonsil
pitted look. The tonsilar crypts often contain histologically resembles the palatine tonsil. Its
parenchyma is also made up of lymphoid nodules
dead epithelial cells, lymphocytes, and other
(n) embedded in dense lymphoid tissue (dl). It
cells that have reached the surface by passing also has the same type of epithelium (e), but the
through the epithelium. tonsillar crypts (c) are shallower. H&E x40.

ESTEBAN & GONZ!\LES' TEXTBOOK OF HISTOLOGY

Fig. XI-18. Pharyngeal Tonsil.
The pharyngeal tonsil also
consists of lymphoid nodules
(n) embedded in dense
lymphoid tissue (dl), but the
epithelium that overlies the
organ is respiratory epithelium
(e). H&E x40.

secreting glands open. When a crypt is present,
it contains masses of dead epithelial cells and
lymphocytes. . the pharyngeal tonsil is referred to as adenoid.
The pharyngeal tonsil occupies the central
area of the posterior and superior walls of the
nasopharynx. Its superficial surface is covered
mostly by respiratory epithelium (i.e., ciliated'
pseudo stratified columnar epithelium), but
some areas may be covered by nonkeratinized
stratified squamous epithelium. The
epithelium of the pharyngeal tonsil forms
shallow folds instead of crypts.
The pharyngeal tonsil sometimes becomes
enlarged, especially in young individuals,
to the point of causing varying degrees of
obstruction to nasal breathing. When enlarged,
The tubal tonsils are located in the
nasopharynx near the openings of the
Eustachian tubes (auditory tubes). They
are really mere extensions of the pharyngeal
tonsil, and like the latter, are covered on their
superficial surface by respiratory epithelium.
Tonsils are better developed in children
than in adults. They start to involute at about
the age of 15 years. In old people, they are
largely atrophied.

LYMPHOID TISSUE _
 .' ..:.i ::
*~* "" ..
"':-(~~~""

Chapter XII

Respiratory System

.
.

H
uman cells utilize oxygen (0) and
produce carbon dioxide (C02) in
carrying out their metabolic processes.
To deliver oxygen and eliminate carbon dioxide
from all the cells of the body, exchange of gases
has to take place between blood and the cells
(internal respiration), and blood and inhaled
air (external respiration),
Internal respiration occurs in all tissues of
the body but external respiration-the function
of the respiratory system-occurs only in the
lungs, specifically, across the ultra-thin blood-
air barrier that separates the blood in the
capillaries from the air in the air sacs (alveoli)
in these organs. Blood is brought to and from
the lung capillaries by the arteries and veins of Fig. XII-1. Component Organs of the
Respiratory System
the cardiovascular system while atmospherih
air is brought to and from the lung alveoli by the
conducting portion of the respiratory system Nose
(i.e., structures that serve as passageways for air
The nose is a hollow organ whose cavity
to ana from the alveoli).
is divided into two irregularly-shaped spaces
(nasal cavities or fossaej by a common
COMPONENT ORGANS OF cartilaginous wall (nasal septum). Each nasal
tHE RESPIRATORYSYSTEM cavity is bounded anteriorly by an orifice
(anterior naris; nostril), and posteriorly, where
The respiratory system consists of the
it is continuous with the pharynx, by another
paired lung~ (left and right) and the organs that
orifice (posterior naris).
conduct air to and from the lungs, and which,
from the most external to the most internal, The nasal septum forms the medial wall
include the nose, pharynx, larynx, trachea, of each nasal cavity while the lateral wall of
and main bronchi. each is occupied by three shelf-like structures,
 cribriform plate
of ethmoid bone

~opening of
.auditory tube

vestibule

hard palate

soft palate
Fig. XII-2. Nose. The
diagram shows the
lateral wall, roof, and
floor of the nose.

the superior, middlet and inferior nasal nasal cavity at the nostril. The hairs of the skin
turbinates or conchae.s that lines the vestibule are coarse and stiff.They
act as a gross filter for inhaled air.
The framework of the walls, roof and floor'
of the nose is formed partly by bone and partly' . The epithelium of the nasal mucosa is the
by hyaline cartilage. Externally, the nose is respiratory epithelturh, except in two areas:
covered by skin while internally, itis lined by at the junction of the vestibule and the rest of
mucous membrane (mucosa), except at the the nasal cavity,where there is a narrow band of
vestibule (i.e.,the spaceor cavityat the entrance nonciliated cuboidal or columnar epitheliumjs
of the nose) which is lined by skin. and, at the roof of the cavity and adjacent areas,
which are lined by olfactory epithelium!
Incidentally, the term mucosa refers to the
moist lining not only of the nasal cavities but The lamina propria of the nasal mucosa
also of the luminal surface of the respiratory, contains mucous and serous glands as well as
digestive, and genitourinary tracts. It consists small collections of MALT. In the area of the
of an epithelium and an underlying loose nasal turbinates, it likewise contains extensive
connective tissue layer called lamina propria~ venous plexuses.
In some segments of the respiratory, digestive, The secretion of the glands in the lamina
and genitourinary tracts, the mucosa has a propria and of the goblet and serous cells in the
third layer that is made up of smooth muscle epithelium keeps the nasal cavity moist while
fibers (collectively referred to as muscularis the venous plexuses serve to warm the air that
mucosae) that separate the mucosa from the passes through the nose.
underlying tissue. The lamina propria of the nasal cavity is
The skin that covers the external surface continuous with the underlying periosteum
of the nose is provided with numerous sweat in areas where it overlies the bone, and with
glands, large sebaceous glands, and fine perichondrium in areas where it overlies the
hairs. It is reflected into the vestibule of the cartilage.

RESPIRATORY SYSTEM ~

cilia gobl&tcells
Fig. XII-3. Respiratory Epithelium. Note that
this type of epithelium-which lines the bigger
air passages-is ciliated pseudostratified with
many goblet cells. Trachea, H&E x200.
Respiratory Epithelium
Respiratory epithelium refers to the
c il'i at e d pseudo stratified columna'
• epithelmm that lines not only the greater part
of the nasal cavities but also many segments
of the conducting portion of the respiratory
system. Six cell types, only some of which can
be distinguished in LM preparations but which
are distinguishable in electron micrographs,
comprise the respiratory epithelium. These
~ cell types, which all rest on the basal lamina,
are as follows: 1) ciliated col~mnar cell-the
most abundant cell type which has up to 300
cilia on its free surface that beat towards the
nasopharynx allowing mucus and particulate
matter that may have been caught in it to be
brought to the pharynx from where they are
either swallowed or expectorated; 2) goblet
cell, 3) br'ush cell-a columnar cell, which
has nume-rous microvilli (instead of cilia) on
its free surface and which is probably a sensory
cell because it is associated with afferent
nerve endings; 4) serous cellLa columnar,
nonciliated cell that has dense apical granules
and whose watery secretion is less viscous
than that of the goblet cell; 5) basal cell-a
short and rounded cell that does not extend
into the epithelial surface and which probably
ESTEBAN & CiO\iZ/\L.ES' TEXTBOOK OF HISTOLOGY
serves as stem cell for the other cell types of the
epithelium; and 6) granule cell (Kulchitsk
cell)-a small cell that is functionally similar
to the neuroendocrine cells that are present
in the epithelium of the digestive tract. The
granule cell looks like a basal cell except for the
presence of numerous electron-dense secretory
granules in its cytoplasm that contain a
hormone and/or cathecolamines that, when
released, probably help regulate the function
of the secretory cells.
Olfactory Epithelium (Organ of
Olfaction)
The epithelium at the roof of the nasal
cavity and over the superior turbinate and
adjacent parts of the nasal septum is referred
to as olfactory epithelium because it contains
the receptors forthe sense of smell. Grossly,the
,
olfactory epithelium is yellowish brown. Thus,
the small area (about 5 em") that it occupies
stands out from its pinkish surroundings that
are lined by the respiratory epithelium.
The olfactory epithelium is a tall (about
60 ~m thick) .pseudostratified columnar
e,pjtqeJi:um. It has no goblet cells and its
basement membrane is indistinct.
Bowman's
glands
°0
Fig. XII-4. Olfactory Epithelium. The three types
of cells that make up the epithelium are shown
in the illustration. Also shown are the Bowman's
glands in the lamina propria.
 olfactory
cells
Fig. XII-S. Olfactory Epithelium. Note the tall, ciliated pseudostratified
The cell types that comprise the epithelium are labeled. H&E x400.
~- N\\ACOS~ ~SSOC'C\.IE.D L~rnPt-lolD l1SSU.~
The t_1:W;kIamina propriafdeep in the
epithelium contains ~ALT and is richly
supplied with blood and lymphatic vessels. It
also contains the olfacto.rY, glands (glands
of Bowmarr), branched tubuloalveolar glands
whose serous secretion moistens the epithelial
surface and serves as solvent for odiferous
substances. QPI~E.ROt)$: G.'VIv'\G OFF Q . .
!>t\'l€I.L, E'OpeClc:n,I.:;, o, OIS\\"'Cnv~
Three types of cells that all rest on the same
basal lamina comprise the olfactory epithelium:
'sustentacular cefl, o!(~~JQ r y.&eIVand ~oUal
cell.
The sustentaculaf or supporting cells
are tall, slender cells that are broad at their
apices and narrow at their bases. Their apical
surface contains numerous, long, and slender
microvilli that are bathed with mucus. Their
nuclei are ovoid and are located a little above
the center of the cells. Their cytoplasm contains
a small Golgi complex, numerous sER, and
lipofuschin granules that are responsible for the
yellowish brown color of the olfactory area. The
sustentacular cells probably provide structural
and functional support for the olfactory cells.
The olfactory cells are spindle-shaped,
bipolar neurons that lie between the
sustentacular cells. Their round nuclei are
situated between the level of the nuclei of the
sustentacular cells and basal cells.
nature of the epithelium.
The dendrite of the olfactory cell passes
between two adjacent sustentacular cells
to terminate in a small bulbous expansion,
the olfactory vesicle, on the surface of the
epithelium. Radiating from the olfactory vesicle
are 6 to 10 fine, hair-like processes (Q!factor~
cilia). The olfactory cilia are very long. They are
structurally similar to the cilia of other ciliated
cells but are non-motile. They lie flattened on
the surface epithelium embedded in the surface
mucus layer. They are probably the actual
receptor elements of the olfactory cell.
The ~~ C9rfaCtorY~Q~r,x£Jiber) of the
olfactory cell, which is on the basal end of
the cell, is unmyelinated and about 0.2 ~m
in diameter. It travels inward into the lamina
propria where it meets the olfactory nerve fiber
of the other olfactory cells. The olfactory nerve
fibers then form many small nerve bundles
(fila olfactoriaj olfactory_nerves) that enter
the cranial cavity through perforations in the
(tiB· ffor.m plate of the ethmoid bone to
terminate in the olfactory bulH=---aknob-like
structure on the inferior side of the brain that is
made up of neurons that process and transmit
olfactory stimuli to the olfactory cortex of the
brain.
The asal cells are small, rounded or
conical, deeply-staining cells that occupy the
RESPIRATORY SYSTEM ~

Fig. XII-7. Epiglottis. This higher magnification
photomicrograph of the epiglottis demonstrates
the epithelium (e) , lamina propria (lp) and its
glands (g), and its framework of elastic cartilage
(ec)enveloped by a perichondrium (p). H&E x100.
area between the bases of the sustentacular
cells and olfactory cells. They have branching
cytoplasmic processes while their nucleus is
dark and ovoid. Basal cells are stemcells that
can differentiate into sustentacular and perhaps
even olfactory cells.
Paranasal Sinuses
The paranasal sinuses) which are named
according to the bone where they are located) i.e.,
£i:9nta1'Jtn:a illaJ:1Y)
etllmoIdal) and splie_noidal)
are cavities in some of the bones of the skull
that arise during embryonic development as
ESTE3!\N & CONZ/\LES' TEXTBOOK OF HISTOLOGY
Fig. XII-6. Epiglottis. The epiglottis is made up of
an elastic cartilage (ec)which forms the framework
and a mucosa that consists of an epithelium (e)
• 1
and a lamina propria (lp). The epithelium on the
anterior surface and upper half of the posterior
surface of the epiglottis, as in photomicrograph,
is nonkeratinized stratified squamous, but
elsewhere, it is respiratory. H&E x40.
invaginations of the nasal mucosa that retain
their connections with the nasal cavity into
adulthood.
The walls of the paranasal sinuses are
lined by mucous membrane that consists of a
rs..~,pirato.ry epithelium that-compared to
that of the nasal cavity-is thinner and contains
fewer goblet cells) and a lamina propria which
contains very few glands and which merges
with the periosteum of the underlying bones.
The direction of the ciliary movement of the
ciliated epithelial cells in the paranasal sinuses
is towards the nasal cavity.
The paranasal sinuses make the face lighter
by reducing its bony mass. They also serve as
resonating chambers for speech.
Pharynx
_ The Irp'har~ is a funnel-shaped'
fibromuscular tube that extends from the base
of the skull to the level of the hyoid bone where
it is continuous with the esophagus. It is a tube
that is common to the digestive and respiratory
systems.
From above downward-s) the pharynx
is successively behind the nasal cavity
nasopharynx)) oral cavity (oropharynx), and
the larynx (laryngop~ary~x).
The wall of the pharynx is similar to that of
the other segments of the digestive tract (see the
chapter on digestive system) in that it is made
up of four histologic layers. In the pharynx)
however) the histologic layers are not as distinct
and as typical as in the other segments of the
digestive tract.
Incidentally, from the luminal surface
outwards) the histologic layers that make up

connective tissue.
The m:ucosa-~ ~~flre phar ynx consists
only of an epithelium and a lamina
propria. The epitheliujjj is nonkeratinized
stratified squamous in the oropharynx and
laryngopharynx: In. t4~ g~~o.l?~~ryn~,it is
respiratory except in areas that are subjected
to friction by the passage of food (e.g.,over the
posterior edge of the soft palate) where it is
nonkeratinized stratified squamous, and in the
roof where it is sometimes stratified columnar
ciliated. The'PlamTiJ.a -J2foilda contains glands
which are pure mucus in the areas that are lined
by stratified squamous epithelium but mixed
in the areas lined by respiratory epithelium. It
also contains some elastic fibers and is rich in
MALT. In fact, as mentioned in the chapter on'
the lymphoid system, aggregates of lymphoid
nodules that are embedded in dense lymphoid
tissue are present in the lamina propria of the
posterior wall of the nasopharynx (plla'ryngccrl"-"
.on.s-il9, lateral wall of the oropharynx
("-p~alanl1e to.!tstID,and around the opening
of the Eustachian tubes in the nasopharynx
(t:t;tbattonsil,.
A submucosa exists in the pharynx in only
two areas, the lateral wall of the nasopharynx
and terminal portion of the laryngopharynx. It
is delineated from the lamina propria by a dense
layer of elastic fibers.
The submucosa, or lamina propria (in the
areas where a submucosa is absent) blends with
the connective tissue that envelops the muscle
bundles of the underlying muscularis externa.
The riffis-cUlWris
emma of the pharynx
consists of two layers of named skeletal but
involuntary muscle fibers: an inner layer
where the muscle fibers are longitudinally-
arranged, and an outer layer where the muscle
fibers are circularly (or obliquely) -arranged.
larynx (Voice Box)
The larynx connects the pharynx to
the trachea and serves an important role in
phonation.
The framework of the larynx is formed
by three unpaired (i.e., tli!iOid, cricoid, and
epiglottic) and three paired (i.e.] ~ta....$'
cuneiform, and arytenoid) cartilages. Of
these cartilages, the arytenoids (exceptfor their
tips which are elastic), thyroid, and cricoid are
Ilyaline, whereas the epiglottic,corniculate, and
cuneiform are el~stic.
The laryngeal cartilages are held together
by ligaments and membranes. They also serve
as attachments for two groups-extrinsic and
intrinsic-of named skeletal muscles. The
extrinsic muscfes connect the larynx to the
surrounding structures. During deglutition,
they raise the larynx. The °hhiosic muscles,
on the other hand, originate and insert within
17.'"'"-:-:-~---:-------..._~-.---,o;-:--:---:-:--~
Fig. XII-B. Larynx. The section is taken from the
part of the organ lined by respiratory epithelium
(e). Note the laryngeal glands (g) that are
embedded in the lamina propria (Ip) which is well
supplied with MALT. The lamina propria blends
with the muscular layer (m). H&E x40.
RESPIRATORY SYSTEM ~
Fig. XII-9. Vocal Cords. The photomicrograph 't' .

shows the true and false vocal cords. In this

specimen, they are both lined by nonkeratinized
stratified epithelium. The false and true vocal
cords are separated by a space, the laryngeal
ventricle. which has pouch-like invaginations
called ventricular recesses. In the lamina propria,
glands are present only in the false vocal cord. In
the true vocal cord, the lamina propria contains
numerous elastic fibers that comprise the vocalis
ligament while deep in the lamina propria are
muscle fibers that comprise the vocalis muscle.
Larynx, H&E x40.
recess

the larynx. They open and close the rima"'ofJ

the-glottjs (i.e., the opening between the vocal
cords) and regulate the tension of the :vocal
cords;
The ,i,Iltema sU.-1rf.:ice
of the larynx is lined
by mucous membrane whose epithelium is
respiratory, except over the anterior surface
and upper half of the po~terfor surface of the
epiglottis, the aryepiglottic folds, and the vocal
cords where it is nonkeratinized stratified
squamous. The direction of the ciliary
movement of the ciliated cells in the respiratory
epithelium of the larynx is upwards, towards
the pharynx.
~""=..",,,,=B'r.o_ia of the lar,yngeal
mucosa is thick. It contains numerous elastic
fibers, blood vessels, nerves, and MALT that
exhibit some lymphoid nodules. It also has large
tubuloalveolar glands 1 that are serous
and mucous, mainly mucous. In the epiglottis,
the glands are similar to the minor salivary
glands (see chapter on the digestive tract) ..
The 1affn ~w.nif2sa blends with the
perichondrium of the cartilages, the connective
tissue that envelops the laryngeal muscles, and
the ligaments and membranes that bind the
cartilages together.
• -6ronchial submucosal gTan(ls) are embedded .
lrachea
ventricle
" The wall of the trachea has four histologic
layers: a) mucosa, b) submucosa, c) carfilage
and muscle -layer, and d) adventitia.
The tradleal-mucosa-..... consists of a
respiratory epithelium that has a very thick
Easement membrane and an abundance of
goblet cells and a lamina propria that contains
numerous elastic fibers and MALT that exhibit
occasional lymphoid nodules. In the deep layer
of the lamina propria, the elastic fibers form a
band that separates the lamina propria from the
submucosa.
The tracheal SUbm.ucbsaconsists of loose
connective tissue where numerous mixed,
tubuloalveolar glands (fr.acheal .gland;s;
Deep in the submucosa, comprising
the bulk of the cartila~ .and. muscle layer
are the most characteristic features of the
trachea-16-20 C-Shaped hyaline cartilage
rings that are responsible for keeping the lumen
of the trachea permanently open. The cartilages

ESTEBA[\J & CO[\JZ/\L[S' TEXTBOOK OF HISTOLOGY


4 except in a few aspects. They have smaller
Fig. XII-11. Trachea (Posterior region). The
photomicrograph shows the trachea lis muscle (m)
that bridges the open end of a C-shaped tracheal
cartilage (c).Also labeled in the photomicrograph
are the epithelium (e) and lamina propria (Ip).
H&E x100.
are stacked on top of each other separated
only by narrow intervals. Each is about 4 mm
in height and 1 mm thick. The posterior ends
of the tracheal cartilages are open. Thus, they
form an incomplete ring around the lumen.
Their open ends are bridged by fiDroelast~o
tligamenf_a:_Fld numerous smopth m"ill!EIiJJers
that comprise the tracheali:S--muscJe which, by
contracting, can decrease the luminal diameter
of the trachea.
The tracRe-al"'~aEe,-ti'fia blends with the
surrounding structures.
Fig. XII-10. Trachea. The photomicrograph
demonstrates the layers of the trachea:
respiratory epithelium (e), lamina propria (lp),
submucosa (sm) that contains tracheal glands
(tg), and hyaline cartilage (he)with its enveloping
perichondrium (p). H&E x100. '
Main Bronchi
The fjITIfin'~~l1i (right and left) that
supply the right and left lungs, respectively,
are morphologically identical with the trachea,
caliber, thinner respiratory epithelium, and
fewer submucosal glands. Furthermore, in the
main bronchi, a discontinuous thin smooth
muscle layer, instead of elastic tissue, separates
the mucosa from the submucosa. The cartilages,
instead of being open posteriorly, form
discontinuous rings around the lumen.
Lungs
The right and left lungs are a pair of conical
organs that occupy the greater part of the
thoracic cavity. They are separated from each
other by the eart and other structures in the
mediastinum ..
Grossly, each lung has an apex that
rises to the neck and a base that rests on the
aiapl1rag.D:, three 15of.d-;;-( aIU!,i 0Ji, iriferiof
and posterior), and two surfaces (costal and
m£_dhistinal) .
The cQsJalsUFfaceof either lung is related
to the ribs and the costal cartilages while the
'mediastinal surface is related to the mediastinal
structures. The mediastinal surface presents a
triangular depression called 'hi,los here the
""- -
structures that comprise the root ofllthe lung
enter and leave the organ. The structures that
_. .
comprise the root of the lung include the niaJit
l:h~,one us, pulmonal'-Y artery and Veins,
bronchial -ar feries'and
7 v.eFrts, IYnl.phatic
vessels, and nerv-es. •
The lungs ar:_ _divided by fi~r~es into
I~~s. The rig_htJung lias three lobes while the
l~ft hiIig has two lobes.
RESPIRATORY SYSTEM ~
 Pleura
~ lung, a main bronchus divides and forms
Each lung is enveloped by a double layer
of fibrous tissue called pleura. The outer layer
of the pleura C.p~:iefci:Iliim-ura)adheres to the
thoracic wall while the inner layer (v.isceral
pleura) adheres to the substance ofthe lung.The
parietal and visceralpleurae-arecontinuous with
each other at the root of the lung. Elsewhere,
1---- pleural
cavity
Fig. XII-12. Pleura
they are separated by a space ~"~leuralcavity),
which contains a small amount of serous fluid.
Histologically, the parietal and visceral
pleurae are made up of connective tissue that
has an abundance of elastic fibers and a relative
paucity of cellular elements that consist mainly
of fibroblasts and macrophages. They are richly
supplied with lymphatic and blood vessels and
nerve fibers. Their frees~~e (i.e., surface
related to the pleural cavity) is lined with
[mesothelium whose cellsare responsiblefor the
minimal amount of fluid in the pleural cavity.
Bronchial Tree
Within the lung, the main bronchus
ramifies dichotomously a variable number of
times-often more than twenty. The term
lironcJI'hll free refers to these generations of
branches.
ESTEB/\N& GONZALES'TEXTBOOK OF HISTOLOGY
Almost immediately upon entering the
secoii(faryHfOiiCpi (lobar bronchi).
A secondary bronchus supplies a lung lobe.
Thus, the right lung has three secondary bronchi
because it has three lobes while the eft lung,
which has two lobes, has only two secondary
bronchi.
The secondary br.onchi divide further
into tertiary bronc-Iii Csegmental.bp!..~chi),
of which there are ten in the right lung and
eight in the left. A tertiary bronchus and the
area of the lung that it supplies comprise a
6roncHo~lil onar'j.y~s.eiment.Thus, the right
lung has ten bronchopulmonary segments while
the left has eight.
The tertiary bronchi ramify into a few
generations of progressively smaller bronchi
before giving offbronchioles.
Bronchioles ramify a few times and then
give rise to lobula.?bronchioles. Arlobular
bronchiole suppliesa ~g lol?uTe,ofwhich there
ate 30-60 per bronchopulmonary segment. The
lung lobules are separated from each other by
incomplete fibrous ~,~pa. They vary greatly in
sizeand shape.The Reri herallo1?pkstend to be
pyramidal, whereas the ~trally Js£ate.a ones
are irregular in shape.
__.
The lobular bronchiole enters a lung lobule
at its apex in the company of the interlobular
r.randies (i.e., branches that supply the lobule)
of the ~ul'monary and br()_l!..chia_~ arteries.
Within the lobule, the branching pattern of
these arteries mirrors the branching pattern of
the bronchial tree.
Lobular bronchioles give off ~rm!.Q..al
hronc1:tioles which, in turn, give rise to
respiratory bronchioles.
A few-a-Iveoli andalyeolar ,sa_c~"(i.e.,
clusters of alveoli) arise directly from the wall
of respiratory bronchioles, but respiratory
bronchioles end by giving off~lveolar ducts-
tiny tubes from which numerous alveolar sacs
and alveoli arise.

Fig. XII-13. Lung. This low magnification
photomicrograph shows the visceral pleura (vp)
that envelops the lung and some structures
in the lung parenchyma. These structures
include the alveoli (al), arteries (ar), respiratory
bronchiole (rb), intrapulmonary bronchus (bs),
and bronchiole (be). H&E x40.
Bronchi
In terms of location, bronchi are either
extrapulmonary or intrapulmonary.
PExtrap_ul!tl""Onary'
bronchi refer to the main
~- . c ".'_. ._
bronchi while inJrapulmonary oronchi
refer to all the bronchi that are within the
lung including secondary, tertiary and their
subsequent branches.
The :bigger-intrapui~;ry "bronchi are
structurally similar to the extrapulmonary
bronchi except that in the intrapulmonary
bronchi, the epithelium is lower and the mucous
membrane is thrown into folds, perhaps as a
consequence of the contraction of the smooth
muscle layer.
prominent.
With each succeeding generation of
branches, the intraplilmonary bronchi exhibit
less and less cartilage, progressively lower
epithelium, and gradual loss of goblet cells.
Bronchioles
B!.2..p.~~h1~W,as a rule, are less than
1 mm in diameter. In routine histologic
preparations, they are easy to tell apart from the
intrapulmonary bronchi because their wall has
no cartilage, submucosal gland, or lymphoid
nodule. Bronchioles bifurcate repeatedly
before ending as lolirilar b(iil£liioles that are
so-called because each supplies a lung lobule.
The epithelium of the bronchioles is still
ciliated, but it progressively diminishes in
height and transforms from pseudostratified
proximally to simple columnar and to simple
cuboidal distally. In the larger .ronch·oles,
the epithelium consists essentially of the same
cells that are present in typical respiratory
epithelium except that there are no serous cells
anymore and that the goblet cells are few.
J
Fig. XII-14. Extrapulmonary Bronchus. The
photomicrograph shows a section taken at low
magnification of an extrapulmonay bronchus.
Different components of the bronchial wall are
labeled in the photomicrograph including the
epithelium (e), lamina propria (lp), muscle layer
(rn), submucosa (sm), hyaline cartilage plates (c),
and adventitia (a). H&E x40.
RESPIRATbRY SYSTEM '141
 consists ofloose connective tissue, merges with
the lung parenchyma.

About 5 to 7 terminal bronchioles arise

from a lobular bronchiole shortly after the latter
enters a lung lobule. Termin~l bronchioles are
small tubes that are less than O.Smm in diameter.
They are considered as the last segment of the
conducting portion of the respiratory system.
Their epithelium is simple cuboidal. Itis largely
non-ciliated, although patches of ciliated cells
are still present together with brush cells,
Fig. XII-1S. Intrapulmonary Bronchus. Note
granule cells, basal cells, and Clara cells, which
that the epithelium (e) is respiratory; the smooth
muscle fibers (m) form a distinct layer between are aplenty. The epithelium has no goblet cells.
the mucosa and the submucosa that contains In terminal bronchioles, the smooth muscle
bronchial glands (g); and the hyaline cartilages
(hc) that keep the bronchus patent consist of
fibers that form bands that separate the lamina
several plates instead of a singte cartilage. Lung, propria from the submucosa are still prominent.
H&E x100. .
~: irat6rV~~.J;9J)~ffi9Ies
In the epithelium of the smaller ond:liol~~,
on the other hand, there are no more goblet Two or more respiratory bronchioles are
cells but in addition to the other cells of the given offby each terminal bronchiole.
respiratory epithelium that are still present, Respiratory bronchioles are short (1-4
there is a population of tall cuboidal, slender, mm) tiny tubes. Their wall consists merely of a
non-ciliated cells, called £lara. cells,;. whose simple epithelium and a thin layer of connective
rounded apices possess microvilli that often
jut out of the surface of the epithelium. Clara
cells have dense secretory cytoplasmic granules
that contain surface active lipoproteins that
are evidently similar to pulmonary surfactant
that reduces surface tension. When released,
the content of the granules probably function
either to protect the bronchiolar lining or to
form a non-sticky layer that prevents luminal
adhesion and helps keep the bronchioles patent.
Clara cells are also stem cells that can divide to
replace the other existing cells in the epithelium.
By the way, Clara ~ells in humans are present
only in the bronchioles, but in lower animals
they are also present even in the bronchi.
I _
The l!!llin~PLQpria of bronchioles contains
Fig. XII-16. Bronchiole. The bronchiole (be) has a
loose collections of MALT. It is separated from
relatively thin wall made up of an epithelium (e),
the submucosa by a distinct smooth muscle lamina propria (lp), a smooth muscle layer (mu),
layer where the muscle fibers are irregularly- and a submucosa (sm). Beside the bronchiole is
arranged. an arteriole (ar). Lung, H&E x100.

ESTEG!:'J~& GONZI\LES' TEXTBOOK OF HISTOLOGY

tissue where islands of smooth muscle cells are
embedded. The epithelium is simple cuboidal
initially but becomes simple squamous distally.
It is populated by granule cells,basal cells,brush
cells, and numerous Clara cells. In the bigger
r_es,pii"at~ry
bronchioles, occasional ciliated cells
exist but there are none in the smaller ones.
Fig. XII-17. Distal Air Passages. The section
shows a bronchiole (be) giving off a terminal
bronchiole (tb) which, in turn, gives off a couple
of respiratory bronchioles (rb). Respiratory
bronchioles give off alveoli (a), alveolar sacs (as),..
and alveolar ducts (ad). Lung, H&E x100.
Alveolarducts are thin-walled conical tubes
that are lined by simple squamous epithelium.
Aside from epithelium, the wall of an alveolar
duct also contains scanty connective tissue
.elements where occasional smooth muscle
fibers are embedded.
The ,a:lv" oli and ,itlYe'oT~iJa<rs
that arise
from alveolar ducts are so numerous that their
openings occupy practically the entire wall of
the alveolar ducts. Consequently, in routine
histologic preparations, the wall of the alveolar
ducts is seen as consisting simply of knob-like
structures that guard the entrances into the
alveoli and alveolar sacs.
A/veo/ar Sacs and A/veo/i
Alveoli arise individually or in clusters
known as ~l:veolar:sacs from either respiratory
bronchioles or alveolar ducts.
Alveoli are thin-walled polyhedral sacs that
are 200-250 tlm in diameter. They are open
on one side to allow entry of air from either
a respiratory bronchiole or an alveolar duct,
depending on which tube they arise from.
The alveoli in the two lungs are estimated
to number around 300 million. They are very
closely packed and share with adjacent alveoli
a common wall (interalv:eol.u septum) that
is perforated by round or oval holes (alveolar.
~poresJ-up to seven per alveolus-~re
2-13 tlm in diameter. The pores, which are
usually clogged with pulmonary surfacta:nt,
may serve as alternate routes for passage of air ~ ~
;::~:?:;~
~~:::v:.o:::::~::er::;::%~Ct~:
alveolar macropliagesj and/or as storage of I
pulmonary surfactant.
Fig. XII-18. Alveolar Duct, Alveolar Sacs, and
Alveoli. The section shows an alveolar duct (ad)
as it gives off alveoli (a) and alveolar sacs (as).
Arrows point to the wall of the alveolar duct
which has been reduced to knob-like structures
that guard the entrances into the alveoli and
alveolar sacs. Lungl H&E x400.
Interalveolar Septum
The connective tissue core of the alveolar
septum contains collagen, elastic and reticular
RESPIRATORY SYSTEM _

Fig. XII-19.Interalveolar Septum
fibers, and several types of cells including
mast cells, plasma cells, lymphocytes, and
l$t.~r§Htii1-fibroblasrs~.which differ from
ordinary fibroblasts in that they contain more
actin filaments, suggestive of the possibility
that they are contractile. Also embedded in
the connective tissue core are numerous blood
capillar·ies (pulmonary capiliI~ies).
Epithelial Cells of the Interalveolar
Septum
Two types of cells comprise the simple
squamous epithelium that lines the luminal
surfaces of the interalveolar septum: tt,pet
l""'1l :veo'lar ~2ell t~ne~monocyfe"""typ'e~ I,
p-ulmonllry eprthe l ial cell, and smal~
alveolar 'ceIl?j and type I a veolar cell
(pneumonocyte type II and great alveolar.
cellY. The type II alveolar cells are more
numerous (60% of epithelial cells) than the
type I alveolar cells, but the latter cells are
stretched very thinly in such a way that they
cover 95% of the alveolar surface, while the
ESTEBM..J& CCNZ/\US TEXTBOOK OF HISTOLOGY
pOlmonary
epithelial
cell
-----_ interalveolar
septum
.:~JmQ.flaJ¥
afvoolar
mae.tepl'lage
type I alv~olarceUs account for only 5% of the
epithelial cover of the alveoli.
Theryp'e--ralveolar eel s are less than 0.2
~m thick, except in areas where the nucleus is
present. They form tight junctions jwith each
other and with the type IIalveolar cells.
Fig. XII-20. Interalveolar Septum. The
photomicrograph shows the components of
the interalveolar septum including pulmonary
capillaries(e)that are lined byendothelialcells (ee),
pneumonocytes type I (pt1), and pneumonocytes
type II (pt2). Manyof the capillaries contain RBCs.
Also shown in the section is a solitary dust cell
(de). Lung, H&E x400.
 The!'y e.l al)Veolar cetls rest on a basal
lamina that is supported by a small amount of
connective tissue, except in those areas of the
epithelium that are associated with capillaries
where the basal lamina of the epithelium is in
direct contact-and often fuses-with the basal
lamina of the capillary endothelium.
The ;tYRe~I!alveolar' cells are much larger
than the type I alveolar cells. They occur among
the type Ialveolar cells either singly or in groups
of two or three. They bulge into the alveolar
lumen or occupy niches in the alveolar wall.
The type II alveolar cells are seen in LM
preparations as cuboidal or round cells with a
large, round nucleus and prominent nucleolus.
Their free surface contains short microvilli. Fig. XII-21. Blood-Air Barrier. The blood-air
The most distinctive feature of their cytoplasm barrier refers to the structures in the interalveolar
is the presence of ovoid, membrane-bound septum (ias) that separates the blood in a
pulmonary capillary from the air in the alveolus.
inclusions fJjulellar bodies} which are the
These structures consist of the pneumonocyte
secretory granules f~r'iYlin~nary surfactant, type I (pt1), basal lamina of the alveolar epithelium
a substance that reduces alveolar surface tension (b11), basal lamina of the capillary endothelium
and thus prevents collapse of the alveoli at the (b12),and capillary endothelial cell (ec).
end of expiration.

Pulmonary Alveolar MacroplTages capillary from the air in an alveolus is called

The most numerous cells in the alveoli are
the pulmonary alveolar macrophages, but they
do not form part of the interalveolar septum.
Some are attached to the interalveolar septum
but most float free in the alveolar lumens.
Monocyte differentiate into macrophages
including the pulmonary alveolar macrophages.
- ~----
These pulm_onaYyil'Vedrrrmacrophages vary in
size from IS to 40 ~m. They are avid phagocytes
and comprise the first line of defense of the
lungs. They contain numerous membrane-
bound cytoplasmic inclusions. Most of these
inclusions consist of phagocytosed materials,
mainly dust particles. This is the reason why
pulmonary alveolar macrophages are often
simply called aiist c_ells~
Blood-Air Barrier
The ultra-thin tissue in the interalveolar
septum that separates the blood in a pulmonary
-~
the' htoo(Fair,
.. ~ -'. \.
It consists of the: 1)
D-,~,rrfet7.
pneumOiiOcyte type I (pulmonary epithelial
cell) j 2) Ba_s-al afiiin.~ of the alve ol er
epifhelium; 3) basarramiiia of the capillaey
~ndotheliumj and 4) capillary endotllelfal
Cell. Often, the two basal laminae fuse together
to form a common basal lamina for the epithelial
and endothelial cells.
Blood and Lymphatic Vesselsof the
Lungs
Two sets of arteries-pulmonary and
bronchial-supply the lungs.
The p_uifuOnary'arferIes, which arise from
the pulmonary tr unk; bring venous blood
to the lungs for oxygenation. Their branches
accompany the bronchial tree as far as the
respiratory bronchioles. Then they break up
into capillaries (pUhnOInlry-capiUarie's) that
form a rich network in the interalveolar septa

RESPIRATORY SYSTEM _
where the blood they contain is oxygenated and
the CO2 in their plasma is released.
@xygena1eabIood from the pulmonary
capillaries is collected by venules-tributaries
of the 'pulmonary veins-that run along the
interlobular connective tissue.
The Drorfcl11alarteries, on the other hand,
arise directly or indirectly from the aorta. They
carry oxygenated blood that supplies the wall of
the different segments of the bronchial tree as
far distally as the respiratory bronchioles, the
rest of the lung parenchyma, the pleura, and the
connective tissue of the lung. Much of the blood
from the bronchial arteries is drained by the
pulmonary veins. However, the rest is drained
by the 6J:!ondiial~~eins which, in turn, drain
into th~ az, gas-system ....
There is no communication between the
pulmonary and bronchial arteries except in
their terminal branches.
ESTEBAN& GOND\LES' TEXTBOOK OF HISTOLOGY
In routine histologic sections, the branches
of the two arteries are distinguishable from each
other because the branches of the bronchial
artery are considerably smaller. In addition, they
have thick walls in relation to their lumens.
The branches of the pulmonary artery and
bronchial artery that supply. a lobule enter the
lobule at the apex together with the bronchiole.
Within the lobule, the branching patterns of the
bronchial and pulmonary arteries mirror those
of the bronchial tree.
The vein that drains the lobule joins the
branches of the pulmonary and bronchial
arteries at the apex of the lobule, and from
thereon, the three vessels travel together
proximally as far as the hilus of the lung.
r:r::ymphaticvessels, on the other hand, travel
in the interlobular septa and are continuous
with the bigger lymphatic vessels beneath the
pleura.
 Digestive Tract

D
igestion' (i.e., the breaking down
of ingested food into absorbable
substances), abs.orption of digested
substances, and excretion of undigested
materials are the functions of the digestive
system. It consists of the digestive tract •
(alimentary canal) and the digestive
glands.'
" DIGESTIVE TRACT
Digestion involvesthe mechanical action of
the digestivetract and the chemical action ofthe
substances and enzymes that are se~~etedby the
digestiveglands-which empty their secretions
into the lumen of the tract-on ingested food.
The digestive tract is a long tube that
consists of several segments that differ
morphologically and functionally. From the
most proximalto most distal, these segmentsare
the oral cavity, pharynx, esophagus, stomach;
small intestin'e, and large intestine. Each
segment of the digestive tract has distinctive
histological features that enable it to perform its
assigned functions.
The digestive glands, on the other hand,
consist of the glands that are embedded in
the walls of the digestive tract (which will be
discussed in this chapter) and the accessory'
glands of the digestive system (which will be
discussed in the next chapter). These glands
are located outside the digestive tract but are
connected to the tract by ducts. The accessory
glands ofthe digestivesysteminclude the major
salivary glands, the exocrine portion of the
pancreas, and the liver and gallbladder.'
GENERAL HISTOLOGIC
ORGANIZATION OF THE
The wall of the digestive tract has four
histologic layers. From the most internal to
the most external, these layers are: 1) mucosa
(tunica muco sa: mucous membrane),
2) submucosa (tunica submucosa), 3)
muscularis externa (tunica muscularis),
and 4) adventitia/serosa (tunica adventitial
serosa). These layers exhibit histologic
variations in the different segments and sub-
segments of the tract. They are not well-
delineated in the oral cavity and pharynx but
are well-definedfrom the esophagusto the large
intestine.
Mucosa (Tunica Mucosa; Mucous
Membrane)
The mucosa of the digestive tract has three
components: 1) epithelium, 2) lamina propria,
and 3) muscularis mucosae. The third is absent
in certain segments of the tract.

-
 Fig. XIII-1: General Histological
Epithelium lines the luminal surface of the
entire digestive tract, but the type varies from
segment to segment. In addition, in many areas,
the epithelium exhibits modifications.
Lamina propria refers to the loose
connective tissue layer that underlies the
epithelium. It contains fine blood, lymphatic
vessels, and mucosa-associated lymphoid
tissue (MALT). In the digestive tract, MALT
is referred to as gut-associated lymphoid tissue
(GALT). Also embedded in the lamina propria
of the different segments of the digestive. tract
are numerous digestive glands.
Muscularis mucosae is the term used to
refer to the thin sheet of smooth muscle fibers
that forms the outermost layer of the mucosa.
The muscle fibers form two layers: an inner layer
where the fibers are circularly-oriented and an
outer layer where the fibers are longitudinally-
oriented.
Submucosa (Tunica Submucosa)
The submucosa which lies external to
the muscularis mucosae is made up of loose
ESTEB/\\: & GO\Z/\L.ES TEXTBOOK OF HISTOLOGY
Organization of the Digestive Tract
connective tissue. In general, the connective
tissue is denser, more abundant, and more
vascular than that in the lamina propria. Like
the lamina propria, the submucosa is also well-
supplied with GALT. In some segments of the
tract, the submucosa has digestive glands.
The submucosa of the esophagus, stomach,
and intestines contains autonomic neurons that
form a ganglionated network, the submucous
plexus (of Meissner).
Muscularis Externa (Tunica
Muscularis)
The muscularis externa typically consists
of two relatively thick, smooth muscle coats or
layers. In the inner layer, the muscle fibers are
circularly-oriented while in the outer layer, the
muscle fibers are longitudinally-oriented.
The muscularis externa is mainly
responsible for the mechanical digestion of
food. Contraction of its smooth muscle fibers
mixes, squeezes, and propels food through the
digestive tract.

In the muscularis externa of the esophagus,
stomach, and intestines, there is a population of
cells that act as pacemakers for the contraction
of the smooth muscles. These cells, called
interstitial cells of Cajal (ICC), are located
between the nerves and the smooth muscle
cells. Ices look like and are not distinguishable
from smooth muscle cells in routine histologic
preparations. They can be identified through
the use of methylene blue staining and zinc-
iodide-osmium impregnation. However, their
distinguishing features, vis-a-vis ordinary
smooth muscle, are best appreciated if electron
microscopy and immunocytochemical
techniques are employed. ICes, like smooth
muscle cells, contain actin and myosin filaments,
dense bodies, and numerous mitochondria.
Furthermore, unlike smooth muscle cells,
their intermediate filament is vimentin instead
of desmin, their sER is flattened, and their
mitochondria are smaller.
Between the inner and outer layers of the
muscularis extern a of the esophagus, stomach,
and intestines lies the myenteric plexus (of
Auerbach). Like the submucous plexus (of
Meissner), it is composed of ganglionated
autonomic neurons. The myenteric plexus
(of Auerbach) and the submucous plexus (of
Fig. XIII-2. Lip (Mucocutaneous
Junction). At the mucocutaneous
junction (unlabeled arrow), the
stratified squamous epithelium
(e) of the lip changes from non-
keratinized to keratinized. The
nonkeratinized epithelium lines
m the mucosal surface of the lip
and overlies a lamina propria (lp)
and. submucosa (sm). Deep in
the submucosa is the muscularis
externa (m), made up of skeletal
muscle. The keratinized epithelium,
on the other hand, is really the
epidermis of the skin of the lip. In
the dermis, sebaceous glands (sg),
a hair follicle (hf), and an arrector
pili muscle (apm) can be seen. H&E
x100.
Meissner) widely Interconnect with each other.
Together, they comprise the enteric division
(enteric nervous system) of the autonomic
nervous system. The enteric division is
responsible for regulating the contractions of
the smooth muscles in the muscularis mucosae
and muscularis extern a, the secretions of the
mucosal and submucosal glands, the caliber
of the blood vessels, water absorption and
electrolyte exchange, and many other activities
that occur in the digestive tract.
As mentioned in the chapter on
nervous tissue, the enteric nervous system
has connections with the sympathetic and
parasympathetic nervous systems, but it
functions autonomously.
Adventitia/Serosa (Tunica
Adve ntitia/Se rosa)
The outermost histologic layer of the
digestive tract is made up of loose connective
tissue. In the areas of the digestive tract covered
by peritoneum, this connective tissue is lined
externally by mesothelium. It is referred to as
serosa. Meanwhile, in areas where the tract
has no peritoneal covering, the layer is called
adventitia.
DIGESTIVE TRACT ~

Fig. XIII-3. Lip (Cutaneous Surface). The skin
of the lip is typical thin skin that consists of an
epidermis (e) and dermis (d). In the dermis, a
hair follicle (hf), sebaceous glands (5g) and some
smooth muscle fibers (sm) can be noted. H&E
x100.
The oral cavity or mouth is divided into
two regions: vestibule and oral cavity proper.
The vestibule is the region of the mouth that
is anterior to the teeth and gums (gingivae)
while the rest of the mouth comprises the oral
cavity proper. The roof of the oral cavity proper
is formed by the hard and the soft palates, the
floor mainly by the tongue and the lateral walls
by the cheeks. The oral cavity is continuous
with the pharynx at the faucial isthmus.
In the oral cavity, food is reduced into
smaller bits by the mechanical action of the
teeth, tongue, and muscles of mastication. It
is then acted upon by the enzymes in saliva
before it is passed on to the stomach via the
pharynx and esophagus. Hence, mechanical
and chemical digestion of food starts in the oral
cavity.
Mucosa and Submucosa of the
Oral Cavity
The mucosa ofthe oral cavityis continuous,
with the skin at the margins of the lips and
with the mucosa of the pharynx at the faucial
[STEt3.:\\ f!<: GO\JZA ...ES' TEXTBOOK OF HISTOLOGY
isthmus. It varies in structure and function in
the different areas of the cavity.
I
On the internal surface of the lips and
cheeks,lthe soft palate, and the ventral surface
of the tongue, the mucosal epithelium is
nonker~tinized stratified squamous. The
lamina propria is loose connective tissue that
is richly supplied with blo~d and lymphatic
vessels, nerves, and GALT. On the other hand,
the submucosa-which is not well-delineated
from the ~amina propria because there is no
muscularis mucosae-contains some small,
mucus-secreting, and compound tubuloalveolar
glands that are named according to their
location, e.g., labial glands (in the upper and
lower lips) and buccal glands (in the cheek).
Over the hard palate and gums, the
epithelium is keratinized stratified squamous.
There is no submucosa in the gums and the
midline part of the hard palate. In these areas,
the lamina propria merges with the periosteum
of the underlying bones. In the rest of the hard
palate, however,a submucosa that also contains
mucus-secreting glands, calledpalatine glands,
is present.
Fig. XIII-4. Lip (Mucosal Surface). Note that the
mucosa consists of a nonkeratinized stratified
squamous epithelium (e) and a lamina propria
(lp). Deep in the lamina propria is a submucosa
where the labial glands (g), a lymphoid nodule
(In) and MALT, and smooth muscle fibers (sm)
are embedded. Deep in the submucosa is the
muscularis externa made up of skeletal muscle
(sm). H&E x100.
Muscularis Externa and
Adventitia of the Oral Cavity
In the oral cavity, a muscularis externa
is present only in the lips and cheeks. This
muscularis externa is very atypical because the
muscles that comprise it are named skeletal
muscles that are arranged in various ways.
These muscles are responsible for mastication
and certain facial expressions.
External to the muscularis externa in the
lips and cheeks is subcutaneous tissue (which
can be considered as the adventitia) that binds
the muscularis externa to the overlying skin.
Tongue
The tongue is a muscular organ that plays
an important role in m~stication, deglutition,
and speech. It also contains most of the sense
organs for taste.
Grossly, the anterior 2/3 of the tongue
(anterior tongue) is demarcated from the
posterior 1/3 (posterior tongue) by an inverted
V-shaped shallow furrow known as sulcus
terminalis. The apex of the CCV"is a depression
(foramen cecum) that marks-the site of
ingrowth of the thyroglossal duct from which
the thyroid gland originated.
Fig. XIII-s. Tongue. The sulcus terminalis
demarcates the anterior tongue from the
posterior tongue.
The anterior tongue forms much of the
floor of the oral cavity while the posterior
tongue forms part of the anterior wall of the
oropharynx. The posterior attached portion of
the tongue is called its root.
Histologically, the tongue is made up of
skeletal muscles that a,re overlaid by mucosa
and, on the ventral surface of the organ, by
submucosa. -
The mucosa on the ventral surface of the
tongue (as previously described) is smooth,
but on the dorsal aspect, it is rough. This is
because on the anterior tongue, it forms small
protrusions called lingual papillae while on
the posterior tongue, it forms lumps due to the
presence ofthe lingual tonsils (discussed in the
chapter on lymphoid tissue). -
Fig. XIII-6. Anterior Tongue. Note that the
epithelium on the dorsal surface of the tongue
contains lingual papillae. The epithelium overlies
a lamina propria. In its ventral surface, the tongue
has a submucosa (not apparent in this section).
Deep in the lamina propria or submucosa are the
skeletal muscle fibers that make up much of the
tongue. H&E x40.
DIGESTIVE TRACT 'II
 epithelium lingua!
tons'll

Fig. XIII-7. Posterior Tongue.

Note the lingual'tonsils under the
epithelium and the mucousglands
deep in the tonsils. H&E x40.

The mucosa of the tongue is atypical In humans, lingual papillae are classified
because it is made up of dens~, instead ofloose according to morphology into three types: 1)
connective tissue. filiform, 2) fungiform, and 3) circumvallate.
The filiform papillae are the most numerous
Lingual Papillae of the lingual papillae. They are slender and
The lingual papillae are projections of the tapering, and are found all over the dorsal
lingual mucosa and are confined to the dorsal surface of the anterior tongue.
surface of the anterior tongue. Each lingual
papilla consists of a core of connective tissue
from the lamina propria that is lined externally
with stratified squamous epithelium that, at the
tips of the papilla, is keratinized.

lamina
p.l'Opr'ia

Fig. XIII-9. Circumvallate Papilla. Note the

Fig. XIII-S. Lingual Papillae. The section shows moat that surroundsthe papilla, the taste buds
two types of lingual papillae: fungiform and embedded in the epithelium,andthe von Ebner's
filliform. Tongue, H&E x100. glands in the laminapropria. Tongue, H&E x50.

ESTE3;\1\i& G()I\iZ/\LES' TEXTBOOK OF HISTOLOGY

 In contrast, the fungiform papillae have
flattened surfaces. They are broader than,
and are scattered among, the filiform papillae.
Their connective tissue core contains numerous
capillaries such that the papillae appear as
pinhead-sized red dots to the naked eye. Taste
buds are occasionally present on the surface of
these papillae.
The circumvallate papillae, on the other
hand, are the largest (1 to 3 mm in diameter)
but fewest (only 6 to 14) of the lingual papillae.
They are arranged in a single file along
the sulcus terminalis. They are cylindrical
structures whose base is surrounded by a canal
or moat. Embedded in the epithelium on the
lateral surface of each circumvallate papilla are
200 to 300 taste buds.
Muscles of the Tongue
-
The skeletal muscle fibers that comprise
the bulk of the tongue form bundles that run
across each other in three planes: horizontal,
vertical, and longitudinal. They are attached to
the hyoid bone, mandible, styloid processes of
the temporal bones, soft palate, and pharyngeal
wall.
Glands of the Tongue
The tongue has a variety of glands.
In the lamina propria at the root and lateral
surfaces of the organ are mucous glands that
are histologically similar to the buccal and
labial glands. Embedded in the lamina propria
and muscles in the region of the circumvallate
papillae are serous glands, and the glands of
von Ebner-compound tubuloalveolar glands
whose ducts open into the moat that surrounds
the circumvallate papillae. The secretions of the
glands ofvon Ebner continuously flush the moat
and serve as a medium where substances are
dissolved in order to be tasted.
In the submucosa in the ventral surface of
the apex of the tongue, meanwhile, are mixed
glands, called the anterior lingual glands (of
Nuhn and Blandin) which are structurally
similar to the glands of von Ebner.
Taste Buds
The taste buds, or the organs that are
responsible for the sense of taste, are ovoid
structures that are embedded in the epithelium
of the circumvallate papillae, and occasionally
of the fungiform papillae, epiglottis, soft palate,
palatoglossal arch, and posterior pharyngeal
wall. .
Fig. XIII-10. Taste Buds. This organ consists of
fusiform cells whose apices converge around
a small opening: the taste pore (at unlabeled
arrow). Tongue, H&E x400.
A taste bud is SO to 80 ~m tall and 30 to SO
~m wide. It consists of SO-ISO densely-packed
fusiform cells whose apices converge on a small
opening on the surface of the epithelium called
taste pore.
Under the LM, three types of cells
are distinguishable in a taste bud, namely:
1) sustentacular or supporting cells, 2)
neuroepithelial or gustatory cells, and 3) basal
cells. The sustentacular and neuroepithelial
cells have a lot of similarities. They are both
slender, spindle-shaped cells whose free surface
is covered by microvilli. They, however, differ
because the sustentacular cells are darker-
staining than the neuroepithelial cells. The
basal cells, meanwhile, are round cells that are
located at the base of the taste bud.
DIGESTIVE TRACT 'IiiI1
 adventitia

~_- outer
muscle layer

inner muscle
layer

submucosa

mucosa

esophag~
glandspro~r

Fig. XIII-11. Esophagus

In electron micrographs, not only three but The taste buds on the anterior tongue are
five types of cells are distinguishable in taste subserved by the facial nerve (eN VII) while
buds: types I, II, III, IV, and V. those on the pharynx are subserved by the
The type IV cells correspond to the basal glossopharyngeal nerve (eN IX) and to a
cells of light microscopy. They serve as stem small extent, by the vagus nerve (eN X).
cells for the other cell types. The type V cells
form the outer boundary of the taste bud. But PHARYNX
the distinction between types I, II, and III cells
is not clear yet. The pharynx is the funnel-shaped,
fibromuscular tube that is common to both the
Many of the type I cells extend lamellate
digestive and respiratory systems. Its structure
processes around the types II and III cells.
has been discussed in the chapter on the
They roughly correspond to the sustentacular
respiratory system.
cells of light microscopy. The type II cells
are similar to the type I cells in that they are
tall cells that contain microvilli on their free ESOPHAGUS
surface.Unlike the type I cells,however,they do
The esophagus is the long muscular tube
not have secretory granules in their cytoplasm.
(20 to 25 ern) that serves as passageway for
They most likely represent a subset of the
food from the pharynx to the stomach.
neuroepithelial cells of light microscopy and
are the transducing cells for the sense of taste.
The type III cells are also long cells but the Mucosa and Submucosa of the
cytoplasmic processes on their free surface are Esophagus
much bigger than the microvilli of types I and
In the esophagus, the mucosa and part
II cells.They probably represent another subset
of the submucosa form longitudinal folds
of the neuroepithelial cells. Furthermore, they
that practically obliterate the lumen of the
contain small, dense vesiclesin their cytoplasm
organ. These folds, however, flatten out when
and are in synaptic contacts with the gustatory
swallowed material passes through the organ.
nerve fibers. They are probably involved in the
transmission of information to the nervous The epithelium of the esophagus,
system. as in the oropharynx, is nonkeratinized

ESTEBAN& GONZi\LES' TEXTBOOK OF HISTOLOGY

stratified squamous. At the gastro-esophageal
junction) this epithelium changes abruptly
to the simple columnar epithelium that
characterizes the stomach. This abrupt
transitional area which forms a serrated border
called Z-Iine is clinically important because it
is the most common site of esophageal cancer.
As in all nonkeratinized stratified squamous
epithelium) in the basal area of the esophageal
epithelium are occasional Langerhans cells)
antigen-presenting cells (APCs) that have been
described in the chapter on the skin.
The lamina propria of the esophagus is
richly supplied with GALT. In the initial and
terminal segments of the esophagus) the lamina
propria contains mucus-secreting) simple
tubular glands called esophageal cardiac
glands. They are similar to the cardiac glands
of the stomach.
Fig. XIII-12. Esophagus. H&E x1.00.
The muscularis mucosae of the esophagus
consists of muscle fibers that are mostly
arranged longitudinally. It is very prominent in
the lower portion of the esophagus but very thin
in the upper portion.
The submucosa of the esophagus is thicker
than the lamina propria. It consists of a very
resilient type of connective tissue containing
GALT) numerous elastic fibers) and small blood
vessels. It also contains the esophageal glands
proper which are compound tubuloalveolar
glands that secrete mainly mucus.
Muscularis Externa and
Adventitia of the Esophagus
The two muscle layers that form the
muscularis externa of the esophagus are not
well-delineated from each other. The outer
layer consists of muscle fibers that are mostly
arranged longitudinally while the inner layer
consists of muscle fibers that are mostly
arranged circularly. However) both layers may
have bundles of muscle cells that are obliquely-
or even spirally-arranged.
The composition of the muscularis extern a
of the esophagus is atypical: in the upper third
of the organ) it consists of skeletal muscle fibers;
in the middle third) it is made up of a mixture of
skeletal and smooth muscle fibers; and in the
lower third) it consists exclusively of smooth
muscle fibers. The skeletal muscle fibers in the
esophagus are involuntary.
The outermost histologic layer of the
esophagus is adventitia) except in the short
segment of the organ that is inside the abdomen)
where the layer is serosa.
STOMACH
The stomach is a J -shaped hollow organ
located in the left upper quadrant of the
abdomen. It presents a left or lateral border
(greater curvature), a medial border (lesser
curvature), an anterior surface) and a posterior
surface.
Grossly) the stomach is subdivided into
the cardia) fundus) body) and pyloric region.
The cardia is the portion of the stomach that
immediately surrounds the esophageal orifice
(cardiac or-ifice}, the opening by which the
esophagus communicates with the stomach.
The fundus is the dome-shaped portion of the
organ that is above the horizontal plane of the
esophageal orifice. The body is the continuation
of the fundus inferiorly and comprises the bulk
of the organ. The pyloric region is the tapering
distal portion of the organ that is continuous
with the duodenum. It is divided into the pyloric
DIGESTIVE TRACT 'fa

Fig. XIII-13. Gastroesophageal Junction.
Note the abrupt transition (at arrow) from the
nonkeratinized stratified squamous epithelium
of the esophagus (on the right side of the
photomicrograph) to the simple columnar
epithelium of the stomach. H&E x100.
antrum and pyloric can~l. The demarcation
lines between the gross parts of the stomach are
histologically not well-defined.
The stomach (which has a capacity of
about 1.S liters) is where food is thoroughly
softened, mixed, and converted into chyme.
Chyme is an acidic semifluid mixture of food
and gastric juice-the watery and acidic fluid
that is secreted by various cells of the gastric
epithelium and glands. There is little absorption
that occurs in the stomach apart from short- and
medium-chain fatty acids and alcohol.
Mucosa of the Stomach
The mucosa and part of the submucosa
form grossly visible longitudinal anastomosing
folds or wrinkles called rugae. When these
folds flatten out, as when the stomach is full, the
mucosal surface looks smooth grossly. But even
when flattened out, the stomach mucosa when
examined under LM, does not appear smooth.
At intervals, the surface epithelium which is a
tall simple columnar epithelium invaginates
into the lamina propria to form shallow furrows
or grooves called gastric foveolae or pits.
The gastric pits, which are 2 to 4 mm apart,
ESTESM~& CiONZhLES'TEXTBOOK OF HISTOLOGY
interconnect with each other extensively. It is
at the bottom of these pits that the glands of the
stomach open into.
Two types of cells comprise the surface
epithelium of the stomach: surface mucous
cells and enteroendocrine cells. The surface
mucous cells comprise the vast majority of the
cells in the surface epithelium. They are mucus-
secreting cells. They also produce bicarbonate
ions that neutralize HCI in the stomach.
Among the surface mucous cells are a few
enteroendocrine cells.
The lamina propria of the stomach consists
of loose connective tissue that has few elastic
fibers and few smooth muscle cells, but rich
in GALT. It is in the lamina propria where the
millions of gastric glands that open into the
gastric pits are embedded.
Fig. XIII-14.Histologic Layers of the Cardia of
the Stomach. H&E x100.
In the stomach, the muscularis mucosae is
well-defined. It consists of an inner layer where
the smooth muscle fibers are arranged circularly
and an outer layer, where the muscle fibers are
arranged longitudinally. Some smooth muscle
cells extend from the muscularis mucosae to the
epithelium. Contraction of these muscle cells
perhaps facilitates emptying of the glands. In
some areas, an outermost layer of muscle cells
that are arranged circularly may be present.
 g,astric
pits

Fig. XIII-1S. Cardia of the

Stomach. Observe that the
gastric pits take as much
space in the mucosa as the
cardiac glands. H&E x100.

·;~~_~>}~i.:
~~'-'._9I_lmd!l ~/-t-;.: .: ~
...
, ,
............
""" .. ~~ jP .. .,'" ,"

.- ... """ ......

.; ~...
...
.:-, .........

Glands of the Stomach Fundic Glands (Principal Gastric

Glands)
The glands of the stomach (gastric glands)
are confined to the la'mina propria. They consist The fundic glands are the most numerous
of three types that are named according to and longest of the glands of the stomach. They
their location: 1) fundic (principal gastric) are populated by five types of cells, most of
which are in the fundus and body; 2) cardiac which are difficult to distinguish in H&E
which are in the cardia; and 3) pyloric which preparations: 1) parietal (oxyntic) cell, 2)
are in the pylorus. All three types are simple zymogenic (chief) cell, 3) mucous neck cell,
tubular or simple branched tubular glands 4) stem cell, and 5) enteroendocrine cell.
whose ducts open at the bottom of the gastric The parietal or oxyntic cell is the most easily
pits. Incidentally, in the glands of the stomach identifiable cell in the fundic gland in H&E
and the intestines, the segment of the gland preparations. It stands out because its intensely
that opens into the mucosal surface is called
isthmus. The part that is immediately basal to
the isthmus is referred to as neck while the rest
of the gland is called base.
It is often possible to tell whether a light
microscopic histologic section of the stomach
has been taken from the cardia, fundus and
body, or pylorus. This is done by examining the
mucosa and comparing the depth of the gastric
pits with the amount of space that the glands
in the lamina propria occupy. In the cardia, the
gastric pits take up about the same amount of
space in the mucosa as the glands. In the fundus
and body, the gastric pits are shallow and the
Fig. XIII-16. Fundus of the Stomach. Note that
mucosa is mostly occupied by glands. In the the gastric pits are shallow and that most of the
pylorus, the gastric pits are very deep and the mucosa is occupied by the fundic glands. H&E
glands occupy just a narrow area of the mucosa. x100.

DIGESTIVE TRACT \B

Fig. XIII-17. Pylorus of the Stomach. Note that
the gastric pits and the pyloric glands occupy just
about the same amount of space in the mucosa.
H&E x100.
eosinophilic cytoplasm contrasts sharply
with the basophilic cytoplasm of the cells that
surround it. This cytoplasmic eosinophilia is
due to the presence of numerous mitochondria.
The parietal cell is pyramidal in shape and
has an oval to round nucleus. In electron
micrographs, it is seen to have a very elaborate
system of branching, the tubular invaginations
of the apical plasmalemma. The parietal cells
produce hydrochloric acid (act) which is
responsible for the acidity of gastric juice. They
also produce intrinsic factor, a glycoprotein
needed for the absorption of vitamin B1 in the
terminal region of the ileum.
The chief or zymogenic cells constitute the
great majority of the cells in the fundic glands.
They are especially numerous in the basal
region of the glands. They are more abundant
in the-glands in the body than in the glands
in the fundus of the stomach. In routine LM
preparations, they are seen as low columnar
cellsthat possess strongly basophilic cytoplasm.
They have secretory granules in their cytoplasm
that contain pepsinogen, the precursor of the
enzyme pepsin.
The mucous neck cells are generally
confined to the neck of the fundic glands
~ ESTEBAN& (-jm~ZALES'TEXTBOOK OF HISTOLOGY
though some are also present in the other areas
of the glands. In routine preparations, mucous
neck cells look very similar to the zymogenic
cells except for their basally located flattened
nuclei. Their cytoplasm is slightly basophilic
and contains numerous secretory granules that
have mucin.
The mucus produced by the mucous neck
cells is chemically different from the mucus
that is produced by the surface mucous cells.
In any case, the combined mucus secretion
of the two cell types forms part of the gastric
mucosal barrier (see page 199) that protects
the stomach surface against the effects of Hel
and proteolytic enzymes.
The stem cells, like the mucous neck
cells, are found mostly in the upper region of
the fundic glands. In H&E preparations, their
nucleus is seen to contain a large nucleolus.
Their cytoplasm is intensely basophilic because
of the presence of numerous ribosomes. The
stem cells can differentiate into surface mucous
cells and gastric gland cells (i.e., mucous
neck cells, oxyntic cells, zymogenic cells, and
enteroendocrine cells). They, therefore, play
an important role in renewing the epithelium
of the stomach, most of whose cells have short
lifespans (for example,the surface mucous cells
live only for three days while the mucous neck
cells survive only for a week).
The enteroendocrine cells
(enterochromaffin cells, argentaffin cells) are
hormone-producing cells that are scattered
singly not only in the fundic glands but also ~I
in the other glands-as well as in the surface
epithelium-of the stomach, and the surface
epithelium and glands of the small and large
intestines.They comprise about 1% of the
epithelial cell population of the stomach and
the intestines.
In H&E preparations, enteroendocrine
cells exhibit a light or clear cytoplasm but are
otherwise indistinguishable from the other
cells. They are columnar, ovoid, or pyramidal,
and contain secretory cytoplasmic granules that
are located mostly in the basal part of the cell.
 -:,
..
::
.. -.', r'~
.','~. ~:N".
~ • Of"
Their secretory granules have a strong affinity
for chromic and silver stains, thus, they can be
identified in LM preparations using these latter
two stains.
The enteroendocrine cells produce
hormones. In terms of the number of cells and
hormones produced, the enteroendocrine cells
collectively form the largest endocrine organ'
in the body. The target cells of their hormones
are, however, limited to the cells involved in the
digestive process.
Electron microscopy and special histologic
techniques have revealed that there are up to
30 types of enteroendocrine cells that differ
ultrastructurally and functionally. The types
of enteroendocrine cells in the stomach
include G-cells that secrete gastrin, EC-cells
that secrete serotonin, D-cells that secrete
somatostatin, and enterochromaffin-like cells
(ECL) that secrete histamine.
Cardiac Glands
The cardiac glands are the least numerous
of the gastric glands. They are morphologically
similar to the esophageal cardiac glands that are
found in the lamina propria in the upper and
lower ends of the esophagus.
The cell types present in cardiac glands are
the same as those in the fundic glands. However,
..' Fig. XIII-18. Fundic Glands. In
'.
.,..- .
."
routine preparations, parietal
cells (at arrows) are the easiest
.,..1 to identify among the cells of
'. ~
I~-
these glands because of their
eosinophilia. H&E x400.
in the cardiac glands, the cells are mostly mucus-
secreting. Among the mucus-secreting cells are
some stem cells, a few enteroendocrine cells
that secrete mainly gastrin, and an occasional
zymogenic or parietal cell.
Pyloric Glands
The pyloric glands are shorter but more
coiled than the fundic and cardiac glands. They
are similar to the cardiac glands in that most
of the cells are mucus-secreting. Among the
mucus-secreting cells are a sprinkling of stem
cells, enteroendocrine cells and parietal cells,
but zymogenic cells are usually absent.
Gastric Mucosal Barrier
The resting pH of the stomach is about
4-5. In the presence of food, the zymogenic
cells release pepsinogen which is converted
into pepsin by H+.Pepsin, the enzyme that starts
protein digestion, works best in a highly acidi
environment. Thus, after a meal, the parietal
cells secrete H CI to lower the pH of the stomach
to 1-2. After pepsin has acted on the proteins,
buffers quickly raise the stomach pH back to its
resting stage.
To prevent HCI and the gastric enzymes
from damaging the mucosa of the stomach,
the stomach has a so-called gastric mucosai'
DIGESTIVE TRACT ...
 _,...--"7 IfIIO Pur Ul-~en.

arrier. The barrier has three components:
compact epithelium brought about by the tight
junctions (zonula occludens) that closely bind
adjacent epithelial cells to each other; 2) a layer
of"mucus over the mucosal surface that has a
median thickness of 180 ~m and is produced by
the surface mucous cells and mucous neck cells;
and 3) bicarbonate ions also produced by the
surface mucous cells, that neutralize acids.
Submucosa, Muscularis Externa,
and Serosa of the Stomach
The submucosa of the stomach is rather
thick. It is made up of connective tissue that is
denser than that in the lamina propria. It is well-
supplied with blood vessels, nerves, and GALT.
The muscularis externa of the stomach
is thick and is composed of-three layers. The
i·Q.n~Lmosnaye~onsists 'of obliquely-arranged
1) a smooth muscle fibers. External to this layer is a
layer of circularly-arranged muscle fibers. The
third and outermost layer consists of muscle
fibers that are arranged longitudinally.
The boundaries between the muscle
layers in the stomach are not well delineated.
Furthermore, the layer of obliquely-oriented
muscle fibers is best developed in the cardia and
is often absent along the lesser curvature. The
layer of longitudinally-oriented muscle fibers,
on the other hand, is best developed in the area
of the curvatures and is absent in the pyloric
region.
In the stomach, as elsewhere in the digestive
tract, the m:y~nt~ric plexus of Xuerbach is
between the layers of longitudinally-arranged
and circularly-arranged muscle fibers.
The stomach is enveloped by peritoneum,
hence its outermost histologic layer is serosa.

Fig. XIII-19. Gastroduodenal Junction. The unlabeled arrow denotes the abrupt transitional area
between the stomach (left side of the arrow) and the duodenum (right side of the arrow). Notice that
the mucosa of the stomach exhibits gastric pits (gp) while that of the duodenum forms intestinal villi
(vi).The lamina propria of the stomach contains pyloric glands (pg). Meanwhile, in the duodenum, the
submucosa (sm) contains Brunner's glands (8g). Note further that the lamina propria is demarcated
from the submucosa by a muscularis mucosae (mm). H&E x40.

ESTESAf'--J& (JCNZ/\LES' TEXTBOOK OF HISTOLOGY

 ~--Iamina
submucosa propria
muscularis
villi
mucosae
mucosal
-t"-,w---- submucosa
fold
'--.:....,,;,.;.ei~-- horizontal
muscularis,
mucosae
lamina
propria
Brunner's
glands

Fig. XIII-20. Duodenum

Fig. XIII-21. Valve of Kerckring (Horizontal

Mucosal Folds) in the Duodenum. The Brunner's
SMALL INTESTINE glands in the submucosa are distinctive features
of the duodenum.
The small intestine with an averagelength
of six to ten meters is anchored to the posterior
body wall by means of the mesenfery. It has
three segments. From the most proximal to
the most distal are the duodenum, jejunum,
and ileum. The Ht'i'bctenumforms the first 25
centimeters of the small intestine while the' ,
je)uB'riin comprises two-fifths and the fkum
three-fifths of the remainder of the ~rgan.
The stomach contents (Eliyme, are
intermittently introduced into the duodenum
via the pylOrIc valVie(pyloric sphinctef), a
strong ring of smooth muscle at the end of the
pyloric canal. In the small intestine, digestion is
more or less completed. It is also in this segment
of the digestive tract where most (about 90%) Fig. XIII-22. Horizontal Mucosa Folds. Jejunum,
H&E x40.
of the nutrients from digested food, as well as
water, are absorbed.
Horizontal Mucosal Folds (Mucosal
Modifications in the Mucosa and Folds; Plicae Circulares; Valves of
Submucosa of the Small Intestine Kerckring; Valvulae Conniventes)
The mucosa and submucosa of the small The luminal surface of the small intestine,
intestines exhibit modifications that are when examined with the naked eye, is not
designed to increase the surface area of the smooth but wrinkled. Histologically, these
organ in contact with food that isbeing digested wrinkles are crescentic or circular horizontal
and absorbed. These modifications come in the creases called horizMital ii;uco~al folds that
form of horizontal mucosal folds, intestinal consist of a core of submucosa and an overlying
villi, and microvilli. mucosa.

DIGESTIVE TRACT CUI

 mU'.~t,llarl$ mucosae
- Jejunum, H&E x100.
The horizontal mucosal folds are most
developed in the jejunum. They are absent in
the first part of the duodenum and the distal
half of the ileum.
Intestinal Villi
The mucosa of the small intestine including
that which lines the horizontal mucosal folds
and the area in-between the folds form finger-
like projections called intestinal villi (Sing.
• villus).
•
An intestinal villus consists of a- core of
lamina propria that is enveloped by epithelium.
Aside from the usual loose connective tissue
elements and lymphocytes, the lamina propria in
the core of a villus contains a few longitudinally-
Fig. XIII-23. Intestinal Villus and Crypt of
Lieberkuhn
ESTES.AN& GCND\LES' TEXTBOOK OF HISTOLOGY
Fig. XIII-24. Intestinal Villi
and Crypts of Lieberkuhn.
oriented smooth muscle cells, blood capillaries,
and lymphatic capillaries called JacteaJ!s. A
lacteal is a blind tube that starts near the tip of
an intestinal villus. Lacteals unite with other
lacteals to form Iymphatfc~essels.
The intestinal willi are numerous in the
proximal segments of the small intestine. They
gradually decrease as one goes distally and
they are often absent in the distal portion of
the ileum. The villi also vary in shape as one
proceeds distally, from broad and tongue-like
in the duodenum to narrow and finger-like in
the ileum.
Microvilli
The epithelial cells that comprise the
surface epithelium of the small intestine have
microvilli on their apical surfaces. These
microvilli are identical to those present in other
cells that exhibit them.
Epithelium of the Small Intestine
The surface epithelium of the small
intestine (i.e., Jthe epithelium that envelops
the intestinal villi and lines the area between
the villi) is siI?pJe columnar'. It is populated
by four types of cells: 1) enterocytes, 2)
goblet cells, 3) enteroendocrine cells, and 4)
M-cells. All four cell types contain microvilli
on their luminal surface, but the microvilli are
most numerous on the enterocytes where they
number up to 3,000 per cell. The microvilli They increase in number if there is ongoing
on the epithelial cells are not individually mucosal inflammation (as in C£U!lC ai~'ease)
distinguishable under the light microscope. and in the presence of tumors (as in the case
Collectively, they form a striated area called of certain colorectaJ"'cancers). IELs are also
lir~sh border (striated border) that is about found in the skin and within the epithelial layer
I um thick. that lines the large intestine, biliary tract, oral
The eilt~rocytes comprise the vast majority cavity, upper respiratory tract and lungs, and
ofthe cellsin the surface epithelium ofthe small reproductive tract. The largest population of
intestine. Under the light microscope, they IELs, however,resides within the epithelium of
are seen as tall cclumnar cells whose nucleus the small intestine.
is typically in the basal part of the cell. They
are absorptive cells that are responsible for Lamina Propria and Muscularis
taking up nutrients from the intestinal lumen Mucosae of the Small Intestine
and transporting these substances across the The lamina propria of the small intestine
epithelium to the lamina propria, where they consists of loose connective tissues richly
diffuse into blood and lymphatic capillaries. supplied with blood and lymphatic vessels,
The goblet cells are interspersed among the nerves, and GALT in the form of diffuse
enterocytes. They increase in number as one lymphoid tissue and occasional lymph nodules.
goes distally ill the small Intestine. It also has smooth muscle fibers some of which
are continuous with the muscle fibers, of the
"Enteroendocrine cells are present in the
muscularis mucosae. Aside from lymphocytes,
entire surface epithelium of the small intestine,
a host of cells that belong to the body's defense
but they are few and far between.
system ispresent in the lamina propria including
M-cells (membranouscefls] miC,roIo 'd·' mast cells, plasma cells, and macrophages. It is
cells9 are present only in areas where there is also in the lamina propria where the inte~tinal •
great concentration of GALT in the lamina glands or crypts of Lieber kuhn are embedded.
propria and submucosa, such as 'the ileum. I c
A typical museu aris mucosae rorrns t e
h
M-cells are large cells with relatively few outermost layer of the small intestinal mucosa.
microvilli. Their basolateral cell membrane
forms indentations that are occupied by
Crypts of Lieberkuhn (Intestinal Glands)
lymphocytes. M-cells are antigen-presenting
cells. They rapidly endocytose antigens from The small intestinal epithelium between
the intestinal lumen, transport these in the form adjacentvilli invaginatesinto the lamina propria
ofvesiclesacrosstheir cytoplasm, and exocytose to form simple tubular glands, called intestinal
them into the indentations in their basolateral glands (crypts of Lieberkuhn), that secrete
membrane where lymphocytes are present. a variety of digestive enzymes. The crypts of
Lieberkuhn span the entire thickness of the
Incidentally, among the epithelial cells
lamina propria and their openings are found
in the small intestine are T-cells called
between the villi.
iiitraepitheliaI lymphocytes (IEL). These
lymphocytes are part of intestinal GALT but Five types of cells comprise the crypts of
are functionally different from the lymphocytes Lieberkuhn: 1) stem cells, 2) enterocytes,
in the lamina propria because they do not need 3) goblet cells, 4) Paneth cells, and 5)
any priming, they immediately release their enteroendocrine cells.
cytokines in the presence of antigens. They The s~emcells are the most numerous cells
are concerned with local immunity as well as in the crypts of Lieberkuhn. They occupy the
immunity to new growths in the intestines. middle region of the glands. They are columnar

DIGESTIVE TRACT Cd
 Fig. XIII-2S. Crypts of Lieberkuhns. The crypts
of Lieberkuhn (at unlabeled arrows) are in the
lamina propria which is separated from the
submucosa (sm) by the muscularis mucosae (mm).
The crypts contain many goblet cells (ge). The
cells in the crypts that have eaosinophilic apical
cytoplasm are Paneth cells (Pc). H&E x400.

cells that are noted in routine histologic

sections to be shorter but more basophilic than
mature enterocytes. They are responsible for
. regenerating the surface and glandular epithelia.
~When they divide) they do so to renew their
numbers or to differentiate into enterocytes,
, enteroendocrine cells) goblet cells) or Paneth have basophilic basal cytoplasm and coarse
'~cells. The resulting cells then either migrate eosinophilic granules in their apical cytoplasm.
up the crypts to replace those cells that are on Paneth cells secrete a variety of antimicrobial
the surface epithelium or descend to populate peptides into the intestinal lumen. These
the basal part of the glands. The stem cells peptides destroy enteric pathogens and dictate
proliferate fast because there. is a rapid turnover "the composition of the microbial colonies in
of epithelial cells in the small intestine: the the small intestine. They may also contribute
lifespan of the epithelial cells (except for the to intestinal homeostasis because their
eI?-teroenCl~lne and Panet~cells which can dysfunction is associated with the occurrence
survive for weeks) is approximately four to six of inflammatory bowel disease) specifically
days only. er,oliii'saisease.'

Most of the .ent:erocyteS'in the crypts -,Enteroendocdne cells are present singly
are destined to replace the enterocytes that in the crypts of Lieberkuhn. Like those in the
are on the surface epithelium. Thus) after stomach) the enteroendocrine cells in the small
differentiating from stem cells) they move intestine (i.e., surface epithelium and intestinal
towards the luminal surface. On their way to '''glands) produce gastrin) somatostatin) and
the surface) they divide several times while at serotonin. But in addition) they produce an
the same time) mature. When they reach the assortment of other hormones including
surface epithelium) they are mature enterocytes entercglucagon, cholecystokinin) gastric
that are absorptive in nature. However) while inhibitory protein (GIP») rnotilin, neurotensin,
still in the crypts) they are largely secretory pancreatic polypeptide) and secretin.
in function) secreting a variety of enzymes
that help complete the digestion of proteins) Submucosa of the Small Intestine
carbohydrates) fats) and nucleic acids.
The small intestine has a thick submucosa
The .goDIet cells are similar to their • that is richly supplied with blood vessels)
counterparts in the surface epithelium) but they lymphatic vessels)and GALT. Solitary lymphoid
are relatively few in the crypts of Lieberkuhn. In nodules are common in the GALT that is
general) they are confined to the upper portions ubiquitous in the submucosa of the entire small
of the glands. intestine. In the ileum) however) especially in
The-P'a,ileth cells comprise a population the anti-mesenteric side) the lymphoid nodules
of cells in the basal parts of the crypts of are particularly large and numerous. They form
Lieberkuhn. In routine histologic preparations) aggregates called Peyer's patclfes that extend
they appear as large) pyramidal cells that into the lamina propria. The lymphoid nodules

ESTEBt\\j & GO\iZAL.ES TEXTBOOK OF HISTOLOGY


in the Peyer's patches are longitudinally-
oriented oval bodies that are sometimes
involved in disease proc~sses'such as"ty.pliot
fever where they may ulcerate.
In the small intestine, submucosal glands
are present only in the duodenum. These glands,
called Q!"_!1!l!1er'sglands (duodenal glands),
are compound coiled, tubular, mucus-secreting,
glands. Their ducts open into the bottom of
the crypts of Lieberkuhn and occasionally in
between villi. Aside from secreting alkaline
mucus that helps neutralize the acidity of
chyme, the Brunner's glands secrete epr3erma'il
growth factor (RGF; urogastrone), a hormone
that is secreted in greater amounts by the major
salivary glands.
Muscularis Externa and Serosal
Adventitia of the Small Intestine
The muscularis extern a of the small
intestine is typical (i.e., inner concentric layer
and outer longitudinal layer) but it is thicker i
the proximal than in the distal portion of the
organ.
The most external histologic layer of the
small intestine is serosa, except for the posterior
aspect of the second and third segments of the
duodenum which is not covered by peritoneum
and is thus simply adventitia.
Fig. XIII-26. Submucous
Plexus of Meissner. The
cluster of neurons and nerve
fibers (at arrow) belong to
the submucous plexus of
Meissner. Note, that the
plexus is embedded in the
submucosa (sm)-that is
delineated from the lamina
propia where the crypts of
Lieberkuhn (cL) are-by the
muscularis mucosae (mm).
Colon, H&E x200.
"
LARGE INTESTI E
The large intestine is about 5 feet in length.
It consists of several segments which, from the
most proximal to the most distal, include the
cecum and vermiform appendix, ascending
colon, transverse colon, descending colon,
sigmoid colon, rectum, and anal canal.
Grossly,the large intestine differs from the
small intestine not only because of its generally
bigger diameter but also because of several
other features-its wall forms pouches called
sacculations or haustrae, small sacs of fat-
filled peritoneum called appendices epiploicae
dangle from the external surface of its wall; and
most of the muscle fibers in its longitudinal
muscle layer collects into three bands called
taeniae coli.
By the time ingested food reaches the
large intestine, digestion and absorption have
practicallybeen completed.The only absorption
that normally occurs in the large intestine is that
of water and some electrolytes, a necessary step
in the formation of semi-solid stool.
Mucosa of the Large Intestine
The mucosa of the large intestine is unlike
that of the small intestine. This is because the
large intestine is devoid of horizontal mucosal
DIGESTIVE TRACT Cd

Fig. XIII-27. Brunner's Glands. The Brunner's
glands (Bg) are the only glands present in the
submucosa (sm) of the small intestine. Other
labeled structures in the photornicroqraph are
the villi (vi), muscularis mucosae (mm), and crypts
of Lieberkuhn (cL). Duodenum, H&E x100.
folds (valves of Kerckring), although in the
rectum and anal canal, there are mucosal folds,
albeit not very elaborate: the transverse rectal
folds and rectal columns of Morgagni. The
transverse rectal folds are two or three folds
of the mucosa that are present in the proximal
portion of the rectum. The rectal columns of
Morgagni, on the other hand, are longitudinal
mucosal folds that are present in the anal canal,
the terminal 4 ern of the large intestine.
Fig. XIII-2B. Myenteric Plexus of Auerbach. The group neurons (at arrows) and nerve fibers at the central
area of the photomicrograph form part of the myenteric plexus of Auerbach. The plexus of Auerbach is
in between the inner layer of circularly-arranged muscle fibers (icm) and the outer layer of longitudinally-
arranged muscle fibers (olm) that comprise the muscularisexterna of the digestive tract. Colon, H&E x200.
[S!TB;\f\i & GONZALES' TEXTBOOK OF HISTOLOGY
There are also no villi in the large intestine,
but crypts of Lieber kuhn are present.
Like the small intestine, the surface
epithelium in most of the large intestine is simple
columnar. At the level of the rectal columns of
Morgagni, the simple columnar epithelium
abruptly gives way to a, nonkeratinized
stratified squamous epithelium which becomes
keratinized at the anal verge.
The cell types that populate the surface
epithelium of the large intestine are the same as
those that line the small intestine: enterocytes,
goblet cells, enteroendocrine cells, and M-cells.
Enterocytes that are absorptive in nature
comprise majority of the cells in the surface
epithelium of the large intestine. In routine
histologic preparations, they resemble the
enterocytes in the small intestine, except that
they have shorter microvilli.
The goblet cells in the epithelium of the
large intestine are similar to those found in the
small intestine, but they are more numerous and
conspicuous in the large intestine. They also
increase in number distally.
The enteroendocrine cells, on the other
hand, are fewer in the large than in the small
intestine. Even the hormones secreted by the
enteroendocrine cells are decidedly fewer than
those secreted by their counterparts in the
small intestine. Among the hormones secreted
by the large intestinal enteroendocrine cells
are glucagon, serotonin, somatostatin, and
pancreatic polypeptide.
The M -cells in the large intestine are similar
to those in the small intestine in structure and
location.
The lamina propria of the large intestine is
rich in GALT. It contains more lymph nodules
than its counterpart in the small intestine.
Some lymph nodules are so large that they
 Fig. XIII-29. Colon.

Fig. XIII-30. Histologic Layers of the Colon.

The low magnification photomicrograph shows
the different histologic layers of the colon:
endothelium (e), lamina propria (lp) where crypts
of Lieberkuhn (cL) are embedded, muscularis
mucosae (mm), submucosa (sm), inner muscular
layer (lml), and outer muscular layer (oml) that
comprise the muscularis externa, and serosa (s).
Colon, H&E x40.

unlike those in the small intestine, do not

. contain Paneth cells, except for a few that are
Fig. XIII-31. Mucosa of the Colon. The colon has
occasionally present proximal to the rectum.
a simple columnar epithelium (e) that invaginates The large intestine has a typical muscularis
in places to form crypts of Lieberkuhn (cL) that
mucosae that consists of two thin layers of
invade the lamina propria (lp). The lamina propia
is separated from the submucosa (sm) by the smooth muscle fibers.
muscularis mucosae (mm). Colon, H&{x100.
Submucosaand Muscularis
extend into the submucosa. Like in the small Externa of the Large Intestine
intestine, aforementioned, the lamina propria
The submucosa of the large intestine, like
of the large intestine contains intestinal glands
that of the small intestine, is thick and richly
(crypts of Lieberkuhn).
supplied with GALT. It has no glands.
The crypts of Lieberkuhn in the large
intestine are a little longer than those in the Muscularis Externa of the Large
small intestine. In addition, they have more
Intestine
goblet cells, less enteroendocrine cells, and
the enterocytes are largely non-secretory. The muscularis extern a of the large
Furthermore, the crypts in the large intestine, intestine is atypical because the outer layer of

DIGESTIVE TRACT '&

 cL
. In

Fig. XIII-32. Rectoanal Junction. Indicated by the unlabeled arrow is the abrupt transition
from the simple columnar epithelium (ce) that characterizes the anal canal to the nonkeratinized
stratified squamous epithelium of the rectum (sqe). There is a narrow region (not discernible in the
photomicrograph) between the two types of epithelia occupied by a stratified columnar epithelium.
At intervals, the epithelium in the anal side invaginates to form crypts of Lieberkuhn (cL).The lamina
propria is heavily infiltrated by MALT which includes a lymph nodule (In). H&E x100.

longitudinally-arranged smooth muscle fibers Serosal Adventitia of the Large

does not surround the organ evenly. Instead,
most of the muscle fibers form three grossly
visible, narrow (about 1 em in width each)
longitudinal bands called taeniae coli that are
equidistant from each other around the large
intestinal wall. In between the taeniae coli, the
layer oflongitudinally-arranged muscle fibers is
very thin and sometimes absent. In the rectum,
however, the taeniae coli disappear and the
muscularis externa reverts to the pattern that is
typical of the digestive tract.
Intestine
For the most part, the outermost histologic
coat of the large intestine is serosa because the
greater part of the large intestine is covered by
peritoneum. The layer is adventitia only in those
parts of the ascending and descending colons
that press directly on the posterior body wall.
The serosa of the large intestine contains
an unusually large amount of adipose tissue that

ESTEBAN s CONZ!\LES' TEXTBOOK OF HISTOLOGY

Fig. XIII-33. Vermiform Appendix.
The histology of the appendix is
essentially that of the rest of the
colon. It has a simple columnar
epithelium (e). Its lamina propria
(lp) is heavily infiltrated by MALT
and is mostly occupied by crypts
of Lieberkuhn. The submucosa (sm)
which lies deep in the muscularis
mucosae (mm) is also heavily
infiltrated with MALT and several
lymph nodules (In) are identifiable.
In between the inner muscle layer
(iml) and outer muscle layer (oml)
that make up the muscularisexterna
are neurons and nerve fibers that
comprise Auerbach's plexus (Ap).
The outermost layer is serosa (s).
H&E x40.

forms grossly visible pendulous masses called
appendices epiploicae,
Vermiform Appendix
The vermiform appendix is a short (average
length is 8 ern), narrow (average diameter is
7-8 mm), and worm-like tubular evagination
of the cecum. It is clinically important because
sometimes, it gets infected, a condition called
appendicitis.
The epithelium of the appendix consists
mainly of tall columnar cells with microvilli;
there are few goblet cells and even fewer
enteroendocrine cells. The appendix has short
crypts of Lieberkuhn that may occasionally
have Paneth cells in their basal portion. Its
lamina propria is typically heavily infiltrated
with GALT where lymphatic nodules are
commonplace. Its muscularis mucosa is poorly
developed.
The submucosa of the appendix is thick
and also heavily infiltrated with GALT. Its
muscularis extern a is complete but not well-
developed. Its outermost covering is serosa.

DIGESTIVE TRACT Eli

 Accessory
Glands of the
Digestive System

he term "accessory glands of the digestive muci ns, immunoglobulins, l}::sosomes,

T system" refers to the digestive glands that

form distinct organs outside the digestive
tract, which include the major salivary glands,
proteins, and inorganic ions. It also contains
salivary corpuscles that consist of degenerating
lymphocytes and granulocytes that come from
exocrine portion of the pancreas, liver and the MALT in the oral cavity and tonsils. The
gallbladder. salivary enzyme~ start the chemical digestion
of food. In addition to its role in the digestion of
SALIVARY GLANDS food, saliva also serves as a so vent for gustatory
su stances. Itlikewise facilitates speech It also
Saliv~l1iy'gla: \1s is the collective name for cleans and protects the teeth, tongue, and other
digestive glands whose ducts open into the oral, oral tissues by its continuous flushing action
cavity. They are classified into minor and major and antibacterial activity.
salivary glands according to size and location.
The '- iQ.~-f,'ts~Jivarygbnds are small glands Major Salivary Glands
that are embedded in the lamina{tia"0pria and
sulimgcm:sa of the oral cav.it;y~They have been
discussed in connection with the oral cavity.
The m~j(xuls<~H~aryg1a:iids!on the other hand,
form distinct organs that are located outside the
oral cavity but their ducts empty into the mouth.

Saliva
The salivary glands secrete about 1.S liters
of saliva per day.
 Incidentally, aside from the major salivary
glands and Brunner's gland, EGF-producing
cells are also present, albeit in fewer number ,
in the anterior pituitary gland, glands of die
stomacH, in the bone marrow, eccrine swea
glands, islet of Langerhans in the pancreas,
lactating breas , adrenal medulla, and renal
medu lao .

- . ,~
Duct System of the Major Salivary
~;1,' Glands
•• '. >
The major salivary glands have well-
developed duct system . The ducts that
are located within the lobules are called
Fig. XIV-1. Parotid Gland. The section shows intralobular ducts while those that are outside
several lobules that are separated from each the lobules are referred to as excretory ducts!
other by interlobular connective tissue (iltc). The There are two types of intralobular ducts
section also exhibits an interlobular duct (ild) and (intercalated and striatedj and three types 0
a lobar duct (ld). H&E x40.
excretory duct (interlobular, lobar and main
excretory) .
serves as passageway for the ducts, blood and The intercalated duct is the segment of
lymphatic vessels, and nerves that supply the the duct system that directly drains an acinus
parenchyma of the glands. (alveolus) or a secretory tubule. It has a narrow
The glandular epithelium hat, comprises lumen, although wider than that in the acini,
the parenchyma of the major salivary glands and its wall is formed by a simple squamous •
or low cuboidal epithelium. The epithelial
forms the secretory unit;. The secretory units,
cells of the intercalated duct, according to some
which are in the shape 0 acini (alveoli) and/or
investigators, also serve as stem cells for acinar
secretory tubules occupy the greater part of tlie
and ductal cells.
lobules. Each secretory unit consists of a sing e
layer of cuboidal cells that rest on a basal lamina
and that are arranged around a small lumen
The major salivary glands are primarily
exocrine glands' that produce the' salivary
enzymes, but they likewise have an endocrine
function The acinar cells of the major salivary
glands also secret epidermal growth factor
(EGF" a hormone which is also produced by
the Brunner's glands in the duodenum and
whose function is to stimulate cell growth, id me sc
proliferation, and differentiation; and inhibit
Hel secretion by the stomach. Salivary EGF
plays an important physiological role in the Fig. XIV-2. Mixed Acinus and Intercalated Duct.
The diagram illustrates a mixed acinus that is
maintenance of oro-esophageal and gastric
made up of mucous cells (mc) and serous cells
tissue integrity. It likewise hastens healing 0 (sc) that form a serous demilune (of Gianuzzi).
oral and gastroesophageal ulcers. The acinus is drained by an intercalated duct (id).

ACCESSORY GLANDS OF THE DIGESTIVE SYSTEM ...

Fig. XIV-3. Striated Duct and Serous Acini. The
photomicrograph demonstrates a striated duct
(sd) that is among purely serous acini (sc). The
unlabeled arrow points to a myoepithelial cell of
the duct. Parotid Gland, H&E x400.

A striated dud (secretory duct) is formed

by the union of intercalated ducts. Its diameter
is iggen than that of an intercalated duct
and its wall consists of a simple cuboidal or
columnar epithelium whose cells, when seen
in routine LM preparation, exhibit intense
cytoplasmic eosinophilia and basal striations~
thus the name. The cytoplasmic eosinophilia
of the cells is due to the presence of numerous
mitochondria while the basal striations have before a main excretory duct opens into the oral
been shown by EM to be due to infoldings ofthe cavity,its epithelium becomes nonkeratinized
oasal plasmalemma. Sometimes, in the terminal stratified squamous.
portions of the due s, just before they merge to
form bigger ducts, the epithelial cells lose their Myoepithelial Cells
basal striations. Associated with the secretory units and
The intralobula .ducts (Le., intercalated the intercalated and striated ducts of the
and striated), aside from serving as passageway, salivary glands are flattened stellate cells
also have secretory function -they modify and called myoepithelial cells.
contribute to the composition of saliva. Myoepithelial/cells, which have been
In the connective tissue between lobule previously described, are contractile cells that
(interlobular septa), the str iated' ducts' are lodged between the epithelial cells and
converge and unite to form interlobular ducts. the basal lamina. They have long cytoplasmic
The epithelium that comprises the wall of rocesses that hug the secretory units and the
interlobular ducts is usuall stratified cuboidal wall of intercalated and striated ducts. Their
in the initial segmet;l and stratified columnar contraction helps eject the secretion of the acini
in the proximal segment of the duct. into the ducts.

The interlobular ducts in a lobe drain

into a single lobar duct that is embedded in
the relatively thick connective tissue between
the lobes (interlobar septa). The wall of a
lobar duct consists of a stratified columnar
epithelium.
Lobar ducts fuse to form a main excretory
duct/s that opens/open into the oral cavity.
The wall of a main excretory duct consists of
an epithelium that is enveloped externally b
dense connective tissue that is supplied with
blood vessels and nerves. The epithelium is Fig. XIV-4. Interlobular Duct. Note that the wall
generally stratified columnar but sometimes, of the duct is formed by a stratified cuboidal
it is pseudo stratified columnar. Shortly epithelium. Parotid Gland, H&Ex400.

ESTEBAN& Cjm~ZALES'TEXTBOOK OF HISTOLOGY

Fig. XIV-S. Lobar Duct. The duct is lined by a
striated columnar epithelium. Parotid Gland, H&E
x400.
Parotid Gland
The parotid glands .are the largest of the
salivary glands. They are located, one on each
side of the body, just below and anterior to the
pinna of either ear at the region of the angle of
the mandible.
In the parotid gland, the lobes and lobules
are well-delineated by connective tissue seEta
that contain a significant number of adiposl
€ell . t
Submandibular Gland
(Submaxillary Gland)
The submandibular glands are located, one
on each side of the body, in the submandibular
fossae on the inner aspect of the mandible,
be ow the floor of the oral cavity.
ACCESSORY GLANDS OF THE DIGESTIVE SYSTEM
__ -----
Fig. XIV-6. Parotid Gland Acini and
Myoepithelial Cells. Note that the cells that
comprise the acini (a) are all serous cells. The
unlabeled arrows point to myoepithelial cells.
H&E x400.
The submandibular glands are smaller and
their connective tissue capsule considerably
thinne than that of the parotid glands. Unlike
in the parotid glands, there are very few adipose
cells in the connective tissue stroma of the
submandibular glands.
The submandibular gland is a mixed gland •
but most of its secretory units are serous. The
mucous acini are few and far between and
some of them even have serous demilunes
(of Giannuzzi). In between adjacent"" mucous
cells in demilune-containing acini are narrow
canaliculi that act as passageways for the
secretion of the serous cells to the lumen of the
acinus.
Fig. XIV-7. Submandibular Gland. Note that the
acini that comprise the gland are mostly serous
acini but there are patches of mucous acini such
as the one pointed to by the arrow. H&E x100.
+Jill
------------------------------~
Fig. XIV-B. Serous Demilunes (at arrows).
Submandibular Gland, H&E x400.
The main excretory duct of the
submandibular gland is called Wharton's duct.
It opens into the oral cavity underneath the
tongue, near the midline, beside tli frenulum
lingua.'
Sublingual Gland
The sublingual glands are the smallest 0
the major salivary glands. They are almond
shaped and located on the floor of the mout ,
underneath the tongu (more specifically,
between the mandible and genioglossus
muscle),one to the left and the other to the right
of midline.
The sublingual gland is embedded in
connective tissue but lacks a definite capsule. It
is a mixed gland but most of its secretory units
are mucous acini~ There are very few purel
serous acin . The serous cells of the gland are
mostly found within the mucous acin in the
form of serous demilunes.
The duct system of the sublingual glands
is not as extensive as those of the parotid
and submandibular glands-striated and
intercalated ducts are few.
Each sublingual gland has between eight to
20 main excretory ducts. The smaller of these
ESTEBAN & GONZ!\LES' TEXTBOOK OF HISTOLOGY
Fig. XIV-9. Sublingual Gland. Observe that the
acini that comprise the gland are mostly mucous
acini but there are some serous cells (at arrows)
which, in this section, form serous demilunes.
H&E x400.
ducts are called ducts of Rivinus. Some ducts
of Rivinus open directly into die sublingual
papilla under the tongue while others empty
into the Wharton's due . Two or more of the
bigger main excretory ducts unite to form the
sublingual duct (of Bartholin) that also drains
into t e Wharton's duct.
~NCR· 5
.• The pancreas is a soft, pinkish organ, 12
to 15 ern in length that stretches transversely
across the posterior abdominal wall from
the. duodenum to the spleen, posterior to
the stomach. It weighs between 60 to 140 g
and consists of a head''that is related to the
duodenum, a body that spans the posterior
abdominal waH;and a tail that is related to the
hilus of the spleen. The pancreas is a vital organ
that is both an exocrine and an endocrine gland.
The stroma of the pancreas consists of
a thin indistinct connective tissue capsule'
that envelops the gland; thin connective
tissue septa that divide the organ into poorly;
delineated lobules and serve as passageway
for blood and lymphatic vessels, and nerves;
and, delicate reticular ttssue.'that supports
the structures within the lobules.'The lobules
contain the pancreatic parenchyma that
consists of the: 1) islets of Langerhans which Pancreatic Acini
comprise the endocrine portion of the organsj
and, 2) secretory units (panc-reatic acini) The pancreatic acini, which comprise the
and intralobular ducts which comprise the bulk of the pancreatic lobules, are grape-like
exocrine portion of the pancreas (exocrine clusters of epithelial cells. Each acinus is made
pancreas), up of 40 to SOclosely-packed low columnar or
pyramidal cells that form asingle layer. The
bases of the cells rest on a basal lamina and their
apices surround a narrow lumen.
In H&E preparations, the epithelial cells
are noted to have a round basally-located
nucleus that contains one or more prominent
nucleolf, Their: apical (slilpt~nU(delr)f~ QpJis)D
is eosinophilic because it contains secretory
granules (zymogen granules) while their 6asal
(infranuelecg) q~f0l!lasomis intensely basophilic
because the granular endoplasmic reticulum of
the cells is extensive and their ribosomes are
numerous. The zymogen granules contain th
precursors of numerous digestive enzymes
Fig. XIV-10. Pancreas. The organ is divided The epithelial cells also contain an extensive
into lobules by interlobular septa (ils), where Golgi complex and elongated mitochondria that
interlobular ducts (ild) are located. The lobules sometimes impart a striated appearance to the
are occupied mainly by pancreatic acini, but
basal region of the cells.
among the acini are islets of Langerhans (iL)., ,
H&E x100. The enzymatic components of pancreatic
juice are synthesized in the rER of the acinar
The is.lets ~of Langerhans,;. in H&E cells. They are then packaged and transformed
preparations, appear as small aggregates of. by the Golgi complex into membrane-bound
pale-staining cells that are interspersed among granules (zymogen granules9 that migrate to the
the pancreatic acini and ducts. Each islet is apical region of the cells. At the right time, the
surrounded by a thin layer of fine reticular fiber contents of the zymogen granules are released
and provided with a rich supply of capillaries. by the cells into the lumen of the acini b
There are over a million islets of Langerhan exocytosis.
in the pancreas, but their combined volum
accounts for only 2% of the volume of the gland.
The islets of Langerhans will be discussed
further in the chapter on the endocrine syst~m.
The fxocfiri:'e panG'reas, which account;
for 98% of the volume of the pancrea , is a
CQihponn(rtubuloacfnaf (tuiffiloa' veoIir
sgrons gland that structurally resembles the
major salivary glands, especially the parotid
gland. Its secretion, called pabcreati j1iic~e.,"
1

amounts to 1.2 liters a day and contains

numerous digestive enzym~s that are necessary
for protein, carbohydrate, and fat digestion. Fig. XIV-11. Diagram of a Pancreatic Acinus

ACCESSORY GLANDS OF THE DIGESTIVE SYSTEM +Jii

Ducts of the Pancreas
The secretion of the pancreatic acini drain
into intercalated ducts whose wall consists,
as in.the major salivary glands, of a simple
squamous or simplelow cuboidal epithelium.
The initial segment of the intercalated ducts
of the pancreas often telescopes into the acinus
that it drains. In routine histologic preparations,
the epithelial cells of these intercalated ducts are
seen as pale-staining cells, called centroacinar
cells, in the luminal area of die acini.
There are no striated ducts in the pancreas,
so the intercalated ducts drain into the bigger
intralobular ducts whose wall consists of a
simple cuboidal epithelium, or directly into
the interlobular ducts.
Like in the major salivary glands, the
intercalated ducts as well as the intralobular
ducts of the pancreas, aside from serving
as passageways, also modify and add to the
secretions of the acini.
Fig. XIV-12. Centroacinar Cells and Intralobular
Ducts of the Pancreas. Centroacinar cells (ca)
are cells of the intercalated ducts that telescope
into the lumen of the pancreatic acini. The
photomicrograph also shows longitudinal
sections of two intercalated ducts (icd) that drain
into an interlobular duct (ild). H&E x100.
ESTEBAN& GONU\LES' TEXTBOOK OF HISTOLOGY
Fig. XIV-13. Intralobular Ducts of the
Pancreas. The section shows cross sections of
an intercalated duct (icd) and interlobular duct
(ild) among the pancreatic acini (a). H&E x400.
In the interlobular septa, the intralobular
ducts drain into interlobular ducts. The wall
of the interlobular ducts consists of a simple
.cuboidal or simple columnar epithelium,
which in the larger ducts is surrounded
externally by connective tissue and some
smooth muscle cells. The interlobular ducts
merge to form two excretory ducts: the main
excretory duct (duct of Wirsung) and the
accessory excretory duct (duct of Santorini).
The wall of the excretory ducts consists of a
simple columnar epithelium, which as in the
larger interlobular ducts, is supported externally
by dense connective tissue that contains some
smooth muscle fibers.
The duct of Wirsung usually joins
the common bile duct to form the
hepatopancreatic ampulla (ampulla of
Vater) that then opens into the duodenum.
Sometimes, it opens, separate from (but in
the company of) the common bile duct, into
the duodenum. The duct of Santorini, on the
other hand, sometimes opens on its own in the
duodenum, a short distance from the ampulla
(of Vater). Sometimes, it drains into the duct of
Wirsung.
LIVER
The liver is the largest gland in the body
and weighs about 1.S kg in adults. It is located
right under the diaphragm and fits into the
diaphragm's concavity,aswell as occupies much
of the right upper quadrant of the abdomen.
Grossly, the liver is divided into several lobes
of unequal sizes: right, left, quadrate and
caudate. Like the pancreas, the liver is a vital
organ.
inferior vena cava
Fig. XIV-14. Liver and Gallbladder
The liver is both an exocrine and an
endocrine gland. Its exocrine secretion is
bile~ which aids in the digestive process by
emulsifying fat. The liver secretes as much
as a liter of bile per day. The hormones the
liver produces include thrombopoietin and
erythropoietin'.
Aside from serving as a gland, the liver has
other metabolic functions. It synt esizesplasma
proteins and hormones, processes dietary amine
acids,carbohydrates, lipids, and vitamins befor
tliese are released into the general circulatio
stores carbohydrates in the form ofglycogenan
releasesthe same in the form ofglucos when the
need arises, detoxifies and excretes endogenous
and exogenous toxic substances through bil ,
and eliminates particulate materials from th
blood.
ACCESSORY GLANDS OF THE DIGESTIVE SYSTEM
Blood Supply of the Liver
The liver receives bloo from two big
vessels,the portal vei and the hepatic artery.
These blood vessels enter the liver at the porta
hepatis, a transverse fissure in the undersurfac
of the right lobe of the orga . It is also at the
porta hepatis where the hepatic ducts exit the
live.
The portal vein brings nutrient-fiIle
venous blood from the gastrointestinal tract to
the liver for processing while the hepatic artery
supplies the liver parenchyma with arteria
bloo . Blood from the portal vein accounts for
60%-70% while blood from the hepatic artery
accounts for 30%-40% of the volume of blood
that goes to the liver.
The portal vein and the hepatic artery
ramify extensivelywithin the liver-each 17to
20 generations of branche . Throughout their
course in the liver, the branches of the porta
vein and the hepatic artery and tributaries of the
hepatic ducts accompany each other.
The terminal branches of the portal vein
ana hepatic artery empty into th hepatic
sinusoids. In the hepatic sinusoids, and only in
the hepatic sinusoids,blood from the portal vein
and hepatic artery intermix.
Histologic Organization of the
Liver
The stroma of the liver is made up of
connective tissue while its parenchyma
consists 0 hepatic cells (hepatocytes).
A main component ofthe liver stroma is the
Glisson's capsule that ismade up ofcolla enou
connective tissue rich in elastic fibers. The
Glisson's capsule envelops the lobes of the liver.
It is lined externally by mesothelium except for
the part that is reflected on the inferior surface
of the diaphragm. It is thin in most of the liver
but rather thick at the porta hepati .
At the porta hepafls, connective tissue
elements from the capsule enter the liver to
+Jii
 Fig. XIV-1S. Capsule and Septa
of the Liver. The lobes of the
liver are invested by a capsule
(c) that sends septa (s) into
the organ to divide it, albeit
indistinctly, into lobules. Also
labeled in the photomicrograph
is a central vein (cv) and a portal
area (pa). H&E x100.

form an extensive network of connective tissue a portal triad, lymphatic vessels, and nerves.
septa that divide the liver lobe, albeit often The term portal triad refers to the interlobular
indistinctly, into lobules (hepatic lobules; branches/tributaries of th hepatic artery
liver lobules). To reiterate, the branches of the (interlobular artery; interlobular arteriole),
portal vein and hepatic artery and the tributarie portal vein (interlobular vein; interlobular
of the bile duct, which accompany each other ..venule), and bile duct (interlobular bile duct).
within the liver, course through these septa.
In a hepatic lobule, the hepatocytes are
Within the hepatic lobules,the connective tissue
organizedinto layers,1to 2 cellthic each,called
stroma consists mainly 0 reticular fibers and
hepatic plates (hepatic cords) that anastomose
reticular cells that form a delicate meshwor
free . The hepatic plates are arranged, like the
spokes of a cartwheel, around avein, the central
Hepatic Lobule (Liver Lobule;
vein (terminal hepatic venule), which is at
Classical Hepatic Lobule)
the center of the lobul . The spaces between
A hepatic lobuleis a polygonalstructure that the hepatic plates are occupied oy sinusoidal
is 0.7mm to 2.0 mm in diameter. Itis delineated
from the other hepatic lobules that are adjacent
to it by connective tissue septa (interlobular
septa). As a rule, in three corners of a hepati
lobule is a portal area-a triangular area 0
interlobular connective tissue that contains

Fig. XIV-16. Classical Hepatic Lobule. The

central vein (cv) occupies the center of a hepatic
lobule. The vein drains the hepatic sinusoids (s)
of the lobule which are separated by liver cells
or hepatocytes that are arranged into layers
called hepatic plates (hp).The diagram also shows
a hepatic arteriole (ha), a bile duct (bd), and a
terminal portal venule (tpv) that give off inlet
venules (iv) which drain into the sinusoids.

ESTEBAN & C;ONZ/l,LES' TEXTBOOK OF HISTOLOGY

~ ---- ----------------~~-----------------------------------------------------

capillaries (hepatic sinusoids) which converg
and drain into the central vel . All the blood
that is brought into the liver by the portal vein
and the hepatic artery ultimately goes into-
and intermix within-the hepatic sinusoids.
From the portal area, blood from th
branches of the interlobular vein and interlobular
artery is brought to the sinusoids through th
following ways. the interlobular vein whicli
gives off branches calle terminal portal
venules (perilobular venules) that course
from the portal area to the periphery of the
hepatic lobu e then give rise to branches calle
inlet venules that empty into the sinusoids] the
interlobular artery which courses through the
periphery of the hepatic lobule then gives of
branches, orne of which empty directly into tli
sinusoi while others empty into capillary
plexus that surrounds the He ductule
(cholangiole; terminal ductule) and which
also drains into the hepatic sinusoids.
The hepatic sinusoids, therefore, receive
both venous and arterial blood. They drain,
as previously mentioned, into the central vein
at the center of the lobul . The central vein
is the first part of the venous side of the liv
circulatio . Neighboring central veins unit
to form sublobular or intercalated veins
ACCESSORY GLANDS OF THE DIGESTIVE SYSTEM
Fig. X V-17. Liver Lobules. A
photomicrograph shows several
poorly delineated liver lobules, each
of which has a central vein (cv) at
its center. In some of the corners of
the hepatic lobules is a portal area
(pa) consisting of connective tissue
where the interlobular branches of
the portal vein, hepatic artery, and
bile duct are ~mbedded. H&E x40.
that are generally at right angles to the centra
vein . Sublobular veins unite to form severa
collecting veins which, in turn, merge to
form several hepatic veins that empty into
the inferior vena cava at the posterior hepatic
surface.
Hepatocytes (Liver Cells)
The hepatocytes comprise about 80%
of the cell population of the live. They
perform practically all the metabolic an
secretor (exocrine and endocrine) functions
of the organ. They are polygonal cells that, as
aforementioned, are arranged in layer (hepatic
plates) that anastomose with each other in the
hepatic lobules. Their surfaces are in contact
either with each othe (lateral surfaces) or with
sinusoids (sinusoidal surfaces). The surfaces
of the hepatocytes are provided with microvil
that are numerous on the sinusoidal surfaces
and sparse on the lateral surface . The latera
surfaces of adjoining hepatocytes form tli
bile canaliculi (further described later in this
chapter).
Most hepatocytes contain a single nucleus
but some have two. The nucleus is typically
roun although sometimes polypoid with one
or two prominent nucleo 1. Hepatocytes have
+JIfI

Fig. XIV-18. Portal Area. In three corners of
the polygonal hepatic lobule is a portal area
(PA) containing the interlobuJar branches of the
hepatic artery (ha), portal. vein (pv), and bile duct
(bd). Liver, H&E x100.
abundant, grainy, eosinophilic cytoplasm
that is loaded with numerous organelles and
inclusion : mitochondria, ribosomes, Golgi
complexes, endoplasmic reticulum (smooth
and rough), lysosomes, and glycogen and lipid-
containing granules. ,.
In so far as their endocrine function is
concerned, hepatocytes synthesize most of the
thrombopoietin in the body, although some
amounts of the hormone are also produced
by cells of the kidney. They also synthesize
erythropoieti ,although they account for.only
10%ofthe erythropoietin produced in the body,
90% is synthesized in the kidneys.
The (von) Kupffer cells, on the other
hand, are fixed macrophages that, like all
macrophages, are derived from monocytes. In
routine histologic preparations, they appear as
stellate cells that possess cytoplasmic processes
that are between the endothelial cells and
that sometimes extend into the lumen of the
sinusoids. They have a nucleus that is pale and
vesicular and cytoplasm that is filled with clear
vacuoles, phagocytosed material, lysosomes,
and lipochrome granules.
The Kupffer cells phagocytose bacteria
and particulate materials that are present in
the sinusoidal blood. In addition, like the
macrophages of the spleen, they also remove
worn-out red blood cells from circulation and
free their iron content for recycling.
Lymph Formation and Lymphatic
Vesselsin the Liver
The liver is the source of more than a third
of the lymph that is produced by the body but
there are no lymphatic vessels in the hepatic
lobules, they are only present in the connective
tissue septa.

Hepatic Sinusoids (Liver Sinusoids)

The hepatic sinusoids are lined by a
thin endothelium that has two types of cells,
endothelial cells and (von) Kupffer cells.
The endothelial cells comprise the
fenestrated endothelial cover of the hepatic Fig. XIV-19. Central Vein and Sinusoids. The
hepatic sinusoids (s) in between hepatic plates
sinusoids. They are flattened cells with an (p) drain into the central vein (cv). The sinusoids
centrally located oval nucleus and scant and the central vein are lined by endothelial cells
cytoplasm. (e). Liver, H&E x400.

ESTEB;\\ s GO\lZ/\L.ES' TEXTBOOK OF HISTOLOGY

 Lymph, which is an ultrafiltrate of plasma,
is formed in the liver by diffusion of plasma
across the highly permeable sinusoidal wall
into the space of Disse. The space of Disse
is the narrow gap that exists between the
hepatic plates and the hepatic sinusoids.
It contains some collagenous and reticular
fibers, and microvilli of the hepatocytes. It
also contains the perisinusoidal cells (of Ito).
The perisinusoidal cells which are sometimes
called lipocytes, interstitial cells or stellate
cells look like pericytes. They have numerous
lipid droplets in their cytoplasm that contairl
dissolved vitamin A. They are the body's main
storage site for vitamin A. They also synthesize
components of the stroma and some bioactive
substances such as prostaglandins. Recent
evidence likewise indicate that they, like
hepatocytes, also synthesize erythropoietin,
Fig. XIV-20. Hepatic Sinusoid and Hepatocyte.
The diagram shows a hepatic sinusoid lined by an
endothelial cell and a von Kupffer cell, as well as
several hepatocytes whose lateral surfaces form
bile canaliculi. The space between the sinusoid
and the hepatocytes is the space of Disse.
The space of Disse is continuous at the
periphery of the hepatic lobule with the space
of Mall-the narrow gap that separates the
tissues in the portal area from the amina
limitans (periportal limiting plate). Lamina
ACCESSORY GLANDS OF THE DIGESTIVE SYSTEM
limitans refers to the hepatic plate that forms
an incomplete wall around each portal area.
Somemicrovilli ofthe hepatocytes that form the
lamina limitans project into the space of Mall.
From the space of Mall, lymph is collected
by lymphatic capillaries which start as blind
tubes in the interlobular ~epta.
In summary, lymph is formed by diffusion
of plasma into the space of Disse, It then flows
into the space of Mall before it finally gets
collected by the lymphatic capillaries in the
interlobular septa.
Bile Passages
Bile is synthesized by the hepatocytes
and secreted into the bile canaliculi that are
in between adjoining hepatocytes. The bile
canaliculi are very small tubes (0.5 to 1.5 ~m
in diameter) that interconnect with each other.
A bile canaliculus does not have a wall of its
own. Instead, its wall is formed by invaginations
of the lateral surfaces of two apposed
hepatocytes. On two areas of this apposition
are desmosomes that effectively prevent the
canaliculus from leaking.A fewmicrovilli ofthe
hepatocytes protrude into the lumen of the bile
canaliculus.
Bile canaliculi discharge their bile at
the periphery of the hepatic lobule into the
cholangioles (terminal ductules; canals of
Hering) that drain into the interlobular ducts
in the portal area. The interlobular ducts, in
turn, drain into the hepatic ducts.
The wall of the initial segment of the
cholangioles is formed by a simple squamous
epithelium that becomes simple cuboidal some
distance before the cholangioles drain into
the interlobular ducts. The wall of the initial
segment of the interlobular ducts, on the other
hand, is made up ofa simplecuboidal epithelium
that is replaced by simple columnar epithelium
before the interlobular ducts join either the left
or right hepatic ducts.
+ail
 Extrahepatic Bile Passages (Extrahepatic
Biliary Ducts; Extrahepatic Biliary Tree)
The hepatic ducts, common hepatic duct,
cystic duct, and common bile duct comprise
the extrahepatic bile passages.
The right and left hepatic ducts leave the .
liver at the porta hepatis and then unite to form
the common hepatic duct that later merges with
the cystic duct of the gallbladder to form the
common bile duct.
As previously stated, the common bile duct
is usually joined by the main pancreatic duct (of
Wirsung) to form the hepatopancreatic ampulla
(ampulla of Vater), which then empties into the
duodenum. The opening of the ampulla ofVater
)
into the duodenum is guarded by the sphincter
of Oddi that is made up of circularly-arranged
smooth muscle fibers.
The extrahepatic' bile passages vary in
diameter but their walls are histologically
similar. Their walls have three histologic layers:
mucosa, which consists of a simple columnar
epithelium that has a prominent basal lamina
and a thin lamina propria that contains
simple mucus-secreting tubuloalveolar glandsj
muscularis, which consists of smooth muscle
fibers, but which is rather thin, and serosal
adventitia.
Portal Lobule and Hepatic Acinus
The hepatic lobule that has been described
in the preceding paragraphs is generally
regarded as the anatomical unit of the live and
is otherwise referred to as classical hepatic
lobule. However, for many experts, the concept
of the hepatic lobule is inadequate to completely
explain the functional intricacies of the liver.
Accordingly, two alternate models with regard
to the architecture of the liver have been
formulated: the portal lobule and hepatic
acinus.
There are thus three ways of looking at
the structure of the liver. The organ can be
viewed as consisting of 1) (classical) hepatic
lobules, or 2) portal lobules, or 3) hepatic
ESTEBN\l& GO\lZ/\L.ES' TEXTBOOK OF HISTOLOGY
----- -------------- --_
Fig. XIV-21. Architecture of the Liver. The
diagram illustrates the three concepts used to
describe the architecture of the liver. The liver
can be regarded as consisting of classical hepatic
lobules, portal lobules, or hepatic acini.
acini. The three models are not conflicting
but rat er complementary interpretations of
the architecture of the liver and understanding
them will greatly enhance one's appreciation
of the physiology and pathophysiology of this
complex organ.
Portal Lobule
The concept of the liver being made up of
portal lobules stresses the fact that the liver is
an exocrine glan .
The portal lobule is the triangular structure
that is bounded by three imaginary straig t
lines that join the central veins of three adjacent
classical hepatic lobul s (see Fig. XIV-21). In
the portal lobule, the central area is occupied by
the portal area where the portal triad is. Hence,
within the portal lobule, blood flows from the
center to the periphery while the glandular
secretion (bile) flows from the periphery to the
center of the lobule-a functional arrangement
that is consistent with that of the other large
exocrine glands in the body. The portal lobule
is considered by many authors as the functional
unit of the liver.
---
Hepatic Acinus
The hepatic acinus is a smaller entity han
a classical hepatic lobule or a portal lobule. It is
an ellipsoidal structure that lies between two
central veins.
In the hepatic acinus, the portal are
is in the peripher of the structure and the
interlobular artery and vein in the portal area
are to be viewed as sending branches into tne
substance of t e liver acinus. q'hese branches
of the blood vessels traverse tne central area 0
the hepatic acinus and course perpendicular to
the imaginary straight line that connects two
central veins,
Another way of visualizing the hepatic
acinus is to regard it as consisting of an ellipsoidal
mass of hepatic plates that are aligned around
a backbone that is made up of the branches of
the hepatic artery and portal vein just as these
vessels empty into the sinusoids.
The liver parenchyma that comprise a
hepatic acinus is divided into three regions:
zones I, II and III. Zone I refers to the regi~I?-
that is closest to the blood vessel backbone.
Zone III refers to the region that is farthest from
the blood vessel backbone while zone II is the
region that is sandwiched by zones I and III.
The concept of the liver consisting of liver
acini helps explain the pathophysiology of the
organ. The hepatocytes that comprise zone I,
being nearest to the blood supply, are the first
to encounter blood-borne toxins and are thus
most susceptible to damage by these substances.
In contrast, the hepatocytes that comprise zone
III, being farthest from the blood supply, are the
most susceptible to damage due to anoxia.
.The hepatic acinus is regarded by many
authors as the true anatomical and functional
unit of the liver.
GALLBLADDER
The gallbladder is a pear-shaped, hollow
pouch that lies in a shallow depression
(gaIlliIaa er fossa) on the right edge of the
visceral surface of the liver. Grossly,it consists of
a fundus, a body, and a neck that is continuous
wit tlie excretory duct of the organ, the cystic.
(iuct.1
As a rule, the bile that is produced by the
liver is first broug t to tlie gallbladder where it is
concentrated, aci mea, and temporarily stored.
When stimulated, the gallbladder releases its
bile content into the duodenum

Fig. XIV-22. Gallbladder. The gallbladder has three histologic layers: mucosa which consists of a
simple columnar epithelium (e) and a lamina propria (Ip), a muscularis (m), and a serosa/adventitia
(a/s). The epithelium invaginates into the lamina propria in many areas to form inpocketings called
Rokitansky-Aschoff sinuses (RA). These inpocketings are non-secretory although they may look like
glands. H&E x100.

ACCESSORY GLANDS OF THE DIGESTIVE SYSTEM WI

Histologic Layers of the
Gallbladder
The gallbladder wall has three histologic
layers.. mucosa, muscularis, and serosal
adventitia.
The mucosa consists only ofthe epithelium,
and lamina propria. The epithelium is simple
tall columnar whose cells,in routine histologic
preparations, are seen to have an oval nucleus
that is basally located and an eosinophilic
cytop asm. In electron micrographs, the
epithelial cells exliibit short microvilli on their
luminal surface. .
The lamina propria consists of loose
connective tissue that is devoid of true glands
except in the neck of the gallbladder where
the epithelium invaginates-into the lamina
propria to form simple tubuloalveolar glands
that secrete mucus. However, the gallbladder
mucosa exhibits some epithelial invaginations
into the lamina propria and the muscle layer.
These inpocketings, called Rok itansky-
ESTEBAN& CONz/\LES' TEXTBOOK OF HISTOLOGY
Aschoff sinuses, are not glands; they are simply
iverticuli.
The muscular layer of the gallbladder
consists of bundles of smooth muscle cells
that are oriented longitudinally, circularly
and obliquely, but do not form distinct layers.
Contraction of the smooth muscle cells of
the gallbladder is induced 'by the hormone
cholecystokinin, which is produced by the
enteroendocrine cells of the small intestine.
When the smooth muscles cells contract, the
sphincter of Oddi reacts by relaxing and bile
flows into the duodenum.
The outermost coat of the gallbladder is
serosa over the organ's posterior and inferior
surfaces, but simply adventitia over the surface
of the organ that is related to the liver.
Occasionally, in the connective tissue that
comprises the serosa around or near the neck of
the gallbladder, smal tubular channels that are
not connected to the lumen of the gallbladder
are present. These tubes are called ducts of
Luschka (supravesicular ducts). They are
probably aberrant bile ducts. .
 Chapter XV
rinary Syste
he organs that comprise the urinary
T system are the paired kidneys and
ureters and the unpaired urinary
bladder-and urethra:
The kidneys perform all the functions of
the system. These include homeostasis-the
maintenance of the body's internal environment
(i.e., acid-base balance and water and electrolyte
concentrations) within normal limits by the
excretion of metabolic waste products and excess
water through urine-and the ptoduction of
hormones, notably renin, erythropoietid, and
thrombopoietin, and other biologically active
substances including kinins and calcitrol~
urinary
bladder
Fig. XV-1.Organs of the Urinary System
(calcitriol; 1, 25-dihydroxycholecalciferol,
vitamin D). The kidneys synthesize possibly all
the renin in the body, 90% of the erythropoietin,
and a small amount of thrombopoietin.
The ureters and the urethra only serve as
passageways while the urinary bladder merely
serves as a temporary storage site for urine.
KIDNEYS
The kidneys are located on the posterior
abdominal wall, one on each side of the body.
They are found at the level of the 12th thoracic
to the level of the third lumbar vertebrae where
they are embedded in fat and connective tissue.
They are bean-shaped and reddish-brown and
measures 10 to 12 em long,S to 6 ern wide,
and 2.5 to 3 ern thick. Each kidney has two
surfaces (anterior and posterior), two margins
(medial and lateral), and two poles (superior
and inferior).
The lateral margin of each kidney is convex
while the medial margin is concave. At the
medial margin, there is a vertical fissure, the
hilus, where the renal artery enters and the
renal vein' and ureter leave the organ. The hilus
is also the gateway to the renal sinus-a 2.5
ern deep fat-filled space within the kidney that
contains the blood vessels, nerves, and the renal
pelvis and calyces.

Fig. XV-2. Kidney, Longitudinal Section.
The kidney is enveloped by -a capsule (c). Its
parenchyma consists of a cortex that occupies
the outer region of the organ which contain the
medullary rays (mr), and a medulla occupied by
the renal pyramids (py) that are separated from
each other by renal columns (rc) of Bertin. The
apices of the renal pyramids fit into cup-shaped
structures called minor calyces (mic). The minor
calyces merge to form major calyces (mac)which,
in turn, unite to form the pelvis (pe). The pelvis
tapers into a tube called ureter (u). The renal
artery (ra) and renal vein (rv) are shown as they
respectively enter and leave the kidney at the
hilus.
t in number as they proceed towards the capsule.
The kidney is envelopedby a thin but tough
fibrous capsules This capsule can be easily
stripped off from the organ because very little
connective tissu from the capsule anchors it
to the parenchyma of the organ. At the hilus,
this capsule lines the renal sinus and becomes
continuous with the walls of the calyces.
Regions of the Kidney
In longitudinal section, the kidney exhibits
two distinct regions: cortex and medulla.
The cortex occupies the outer region (i.e., the
area beneath the capsule). It is reddish and contain the distal segments of the collecting
granular. The medulla, on the other hand, tubules. In addition, they also consist of the
which comprises the inner region, is striated. .papillary ducts, into which the collecting
ESTE3M'~& CCNZALES' TEXTBOOK OF HISTOLOGY
The junction of the cortex and the medulla is
delineated by large, arched blood vessels, the
arcuate arteries and veins.
The renal medulla consists mainly of 10
to IS grossly-visible, longitudinally-striated,
conical structures called renal pyramids. These
are separated from each other by the renal
columns (of Berttn) which are projections of
cortical tissue into the medulla. The base of
each pyramid abuts the cortexwhile the aPlixor
renal papifla'points toward the hilus.
At the corticomedullary junction, groups
of parallel tubules that are grossly visible a-s
longitudinal streaks-the medullary rays (of
Ferreirl)-seem to leave the medulla from the
bas of each renal pyramid and traverse the
cortex, The medullary rays are referred to as
pars radiata. There are 400-S00 medullary
rays per pyramid.
A medullary ray consists of the proximal
segments of a group of collecting tubules'. The
distal segments ofthese collectingtubules are in
the renal pyramids.Hence, the term "medullary'
refersto the destination of the tubules.not their
origin. In addition to the collecting tubules, the
medullary rays also contain segments of the
loops of Henle. These latter structures will be
described later in this chapter.
The tubules in the medullary rays diminish
In a short distance from the capsule, they
completelydisappear.
The region in the cortex in between th
medullary rays is referred to as pars convoluta.
(cortical labyrinth). It contains all the renal
corpuscles' and parts of the. renal tubules,'
specificallythe proximal and distal convoluted
tubules. Italso contains blood vessels,minimal
connective tissue, and the arched segments of
the ollecting tubules. These structures will
also be described later in this chapter.
The renal pyramids, as mentioned above,
tubules drain; the segments of the loop of Henle
that dip into the medulla, and blood vessels and
some connective tissue elements. The renal
columns (of Bertin). on the other hand, contain
segments of the loop of Henle, blood vessels,
nerves, and connective tissue elements.
The terminal segments of the papillary
ducts form the renal papilla which fits into a
cup-shaped structure, the minor calyx. Several
minor calyces unite to form 2-3 bigger tubular
structures, the major calyces. The major.
calyces, in turn, unite to form the renal pelvis
which is simply the expanded initial portion of
the ureter.
lobes and lobules of the Kidney
A renal pyramid and the cortical tissue that
overlies its base and surrounds its sides (i.e.,
associated renal column) constitute a renal
lobe. Thus, a kidney has 10 to 15 lobes because
it has 10 to 15pyramids.
A renal lobe is made up of numerous
conical structures called renal lobules that are
partly in the cortex and partly in the medulla.
A renal lobule consists of a papillary duct:
the collecting tubules that drain into it: the
nephrons (i.e., renal corpuscles and renal
tubules) that drain into the collecting tubules,
and the blood vessels and connective tissue
elements associated with the papillary duct,
collecting tubules, and nephrons.
There are no. connective tissue septa that
separate the renal lobules from each other. In
the cortex, the peripheral boundary of the renal
lobules is formed by the interlobular blood
vessels while their center is occupied by a
medullary ray.
Blood Vessels, lymphatics, and
Nerves of the Kidney
The arteries that supply the left and right
kidneys are the left and rtght renal arteries,
respectively. They arise from the abdominal!
aorta. On the average,they deliver about 1.2 L
of blood/min to the kidneys for filtering.
At the hilus of the kidney, the renal artery .
divides into three to five branches that enter
Fig. XV-3. Renal Lobe. The renal lobe is made
up of a renal pyramid. In this section, the pyramid
occupies the whole medulla (me) and the cortical
tissue (eo) that overlies it. The corticomedullary
junction is designated by the arcuate blood
vessels, arcuate artery (ar), and arcuate vein (av).
Groups of parallel tubes that traverse the cortex
form what are called medullary rays (mr). The
tubes that comprise the medullary rays diminish
in number as they approach the capsule (ea).
In the region between the medullary rays are
the renal corpuscles (rc). The apex of the renal
pyramid, called renal papilla (pa), fits into the
cup-shaped minor calyx (me). Kidney, H&E x1.
URINARY SYSTEM ..

the renal sinus where they give rise to a variable
number of interlobar arteries. These arteries
penetrate the medulla, creep between the renal
pyramids, and proceed to the cortex without
giving off any branches. In other words, the
interlobar arteries pass through the medulla via
the renal columns of Bertin.
At the corticomedullary junction, the
interlobar arteries end by giving rise to arcuate
arteries that course parallel to the capsule. The
are no anastomoses among arcuate arteries.
An arcuate artery branches off at right
angles and gives off interlobular arteries-
straight vessels that (as mentioned earlier)
form the outer boundaries of the renal
lobules. The interlobular arteries proceed
ESTEBAN& GOND\LES' TEXTBOOK OF HISTOLOGY
Fig. XV-4. Corticomedullary Junction (A).
In the kidney, the arcuate blood vessels,
arcuate vein (av), and arcuate artery (aa)
occupy the corticomedullary junction. In the
photomicrograph, the cortex-where the renal
corpuscles (rc) are embedded-occupies the
region to the right of the blood vessels while the
medulla occupies the region to the left. Kidney,
H&E x100.
Fig. XV-So Renal Cortex (8). The cortex-the
region of the kidney that is immediately below
the capsule (c}-is characterized by the presence
of the medullary rays (mr) which collectively
comprise the pars radiata. The area between
the medullary rays makes up the pars convolute
which contain the renal corpuscles (rc). Kidney,
H&E x40.
Fig. XV-6. RenalCortex (C).The photomicrograph
demonstrates the same structures shown in the
previous figure-medullary rays(mr) and the pars
convolute (pc) which contain the renal corpuscles
(rc}-at higher magnification. Kidney, H&E x100.
towards, and ultimately supply, the renal
capsule. However, along the way, they give
off afferent arterioles that course parallel to
the capsule and enter the lobules. Within the
lobules, each afferent arteriole breaks up into a
clump of capillaries (glomerular capillaries).
The glomerular capillaries drain into a single
efferent arteriole.
Fig. XV-7. Uriniferous Tubule. In the
schematic diagram, the pink region below
the capsule (e) represents the cortex while
the blue area represents the medulla. The
uriniferous tubule is partly in the cortex
and partly in the medulla. It is made up of
the nephron and the intrarenal ducts. The
nephron consists of the renal corpuscle and
renal tubule. The renal corpuscle is made up
of the glomerulus (g)-where the afferent
arteriole (aa) and efferent arteriole (ee) can
be seen to enter and leave, respectively-and
Bowman's capsule (Be). The renal tubule is
made up of the proximal convoluted tubule
(pet), thick descending limb (tdl) , thin limb (tl)
and thick ascending limb (tal) of Henle, and
distal convoluted tubule (det). The intrarenal
ducts consist of the collecting tubules (et) and
papillary ducts (pd).

Efferent arterioles are short portal vessels.
Most break up to form a peritubular capillary
network that supplies the proximal and distal
tubules, and the segments ofthe loops Henle of
that are around them. Meanwhile,those that are
associated with the juxtamedullary nephrons
(see page 234) break up to form a series of
straight capillaries (vasa recta) that take a,
descending path (descending vasa recta) into
the medulla-which they supply-then loop
and ascend baok (ascending vasa recta) to the
corticomedullary junction.
Venousblood from the capsuleand the outer
cortex drains into the superficial cortical veins
that empty into bigger veins (stellate veins)
found on the surface of the kidney. Venous
blood from the rest of the cortex drains into
the deep cortical veins. The stellate and deep
cortical veins empty into the interlobular veins
that run parallel and close to the interlobular
arteries. The interlobular veins, in turn, drain
into the arcuate veinJ.
Venous blood from the medulla, on the
other hand, drain into the medullary veins
(i.e.,ascending vasa recta). These veins empty
either into the interlobular veinsjust before they
join the arcuate veins or directly to the arcuate
veins. The arcuate veins are tributaries of the
interlobar veins that unite to form the renal
vein which drains into the inferior vena cava.
From the arcuate veins to the renal veins,
the veins accompany their corresponding
arteries.
In the kidney, the lymphatic vessels and
sympathetic nerves follow the pattern of
distribution of the arteries.
Uriniferous Tubule
The term uriniferous tubule is a collective
term for the nephron and the intrarenal ducts
(i.e., collecting tubules and papillary ducts).
It was originally intended to make a distinction
between the structures that produce urine
(nephrons) and the tubes (intrarenal ducts)
that conduct urine to the main excretory ducts,
the initial segments of which are the calyces.
Recent evidence, however, has shown that the
intrarenal ducts are not merely conduits for,but
are also participants, in the formation of urine.
Nephron
The nephron is the functional unitof
the kidney. Each kidney has between 1 to 1.S
million of them. The nephron has two parts:
renal corpuscle and renal tubule. Urine is
formed by filtering blood which occurs in the
renal corpuscle, and then modifying the filtrate
(glomerular filtrate) which occurs largelyin the
renal tubule.

URINARY SYSTEM +Ill

Renal Corpuscle (Malpighian Corpuscle)
The renal corpuscle forms the proximal
end of the nephron. It is a spherical structure
that is about 200 ~m in diameter and has two
components: glomerulus and Bowman's
capsule (glomerular capsule).
The renal corpuscle has two poles:
vascular and urinary. The vascular pole is
where tlIe afferent arteriole enters and the
efferent arteriole leaves the corpuscle. The
urinary pole is where tlie renal tubule begins.
All the renal corpuscles are located in, and
account for the gra_inyappearance of, the cortex.
capillaries (glomerular capillaries) that form 20
to SO highly convoluted loops that anastomose
with each otheu
The glomerular capillaries which comprise
the bulk of the glomerulus are fenestrated.
However, unlike fenestrated capillaries
elsewhere in the body, their fenestrae or pores
are not covered by a diaphragm, but their
endothelial cells rest on a very thick basal
lamina.
The spaces between the glomerular
capillaries are filled with mesangial matrix, an
amorphous material that is similar to basement

Glomerulus
The glomerulus is a ball-like structure made
up of capillaries (glomerular capillaries),
mesangial matrix (mesangium), and
glomerular mesangial cells.
Upon entering the vascular pole of the renal
corpuscle, the afferent arteriole breaks up into

Fig. XV-B. Renal Corpuscle, Vascular Pole.

The parts of the renal corpuscle shown in the
photomicrograph are the glomerulus (g), the
parietal layer of Bowman's capsule (p), .and the
Bowman's space (Bs).Also shown are the afferent
(aa)and efferent (ee) arterioles as they enter and
leave the corpuscle and segments of the renal
tubule-proximal convoluted tubules (pct) and
distal convoluted tubules(dct). Kidney, H&E x400.

Fig. XV-9. Renal Corpuscle,

Urinary Pole. As in the
previous figure, the parts of
the renal corpuscle shown in
the photomicrograph are the
glomerulus (g), the parietal
layer of Bowman's capsule
(p), and the Bowman's space
(Bs) which is continuous with
the initial segment of the
proximal convoluted tubule
(pcv). Kidney, H&E x400.
membrane and serves to bind the loops of the
glomerular capillaries. Itis also in the mesangial
matrix where the glomerular mesangial cells are
embedded.
. The glomerular mesangial cells are most
numerous in the vascular pole of the renal
corpuscle. They are stellate cells similar to
pericytes in that they are contractile and could
influence the luminal diameter of the glomerular
capillaries. Unlike pericytes, however) they are
capable of phagocytosis and have cytoplasmic
processes that crawl between endothelial cells
to reach the capillary lumen. They are generally
credited with helping maintain the glomerula]j
filtration barrier by keeping it free from debris.
They are also responsible for the production of
mesangial matrix.
" called foot processes or pedicels. The pedicels
/capillary walls where they interdigitate with the
Fig. XV-10. Relationship of Podocytes to
Glomerular Capillaries. The pedicels of the
podocytes and the endothelial cells of the
capillaries share a basal lamina. The spaces
between the podocytes comprise the filtration
slits.
Bowman's Capsule (Glomerular Capsule)
The Bowman's capsule is a double-walled
sac that envelops the glomerulu . Its structure
is analogous to an inflated rubber balloon that
has been pushed inwards on one side by the
glomerulus to form a cup.
Both walls of Bowman's capsule consist
of a single layer of flattened epithelial cells.
The inner wall (visceral layer) envelops the
glomerulus intimately while the outer wall
(parietal layer) forms the outer boundary of
the renal corpuscle. Between the visceral and
parietal layers of Bowman's capsule is a narrow
cavity known as Bowman's space (capsular
space)subcapsular space)urinary space).
At the vascular pole of the renal corpuscle)
the parietal layer of Bowman's capsule is
continuous with the visceral layer. Meanwhile
at the urinary pole) it is continuous with the
epithelium of the renal tubule.
The cells that comprise the parietal layer
of Bowman's capsule form a simple squamous
epithelium while those that form the visceral
layer are highly modified and are called
podocytes.
Podoeytes are stellate cells that have
cytopla(mic processes (major processes) that
vary in size and shape. These major processes
branch further and give rise to minor processes
wrap themselves around the glomerular
pedicels of neighboring podocytes. They are the
only parts of the podocytes that rest on the basal
lamina which they share with the endothelial
cells of the capillaries.
Fig. XV-11. Glomerular Filtation Barrier. The
barrier consists of the fenestrated endothelium
(f) of the glomerular capillaries, the basal lamina
(bl) common to the endothelial cells and pedicels,
and the filtration slits between the pedicels (pe)
of the podocytes (p). x1O,OOO.
URINARY SYSTEM WI
 The narrow gaps that exist between the
interdigitating pedicels are called filtration
slits. They are covered by a thin electron-
dense membrane, the slit membrane or slit
diaphragm, which is dotted by very small pores.
Glomerular Filtration Barrier
In the renal corpuscle, a very thin sheet
of tissue called glomerular filtration barrier
separates the blood in the glomerular capillaries
from the Bowman'sspace.This filtration barrier
consists of the 1) fenestrated endothelium of
the glomerular capillaries j 2) basal lamina
that is common to the endothelial cells
and pedicels, and 3) slit membrane (slit
diaphragm) of the filtration slits between the
pedicels.
The fenestrae of the glomerular
endothelium, as previously mentioned, are not
covered by a diaphragm. Their diameter varies
from 60 to 100nm.
The basal lamina, on the other hand, is
330 nm thick. It is much thicker than the basal
lamina of fenestrated capillaries elsewhere in
the body.
The slit membrane, the third component of
the filtration barrier, is about 6 nm thick. The
filtration slit that it bridges is about 2S nm wide.
As blood passes through the glomerular
capillaries, filtration (glomerular filtration)
occurs. In this process, the glomerular filtration
barrier acts as a sievethat allows an ultrafiltrate
of blood (glomerular filtrate) to seep into
Bowman's' space.
Renal Tubule
From Bowman's space, the glomerular
filtrate flows into and traverses the very long
renal tubule where it is modified-most of
the water and solutes are reabsorbed (tubular
reabscrption) and some other solutes are added
(tabular secretion)-before it finally becomes
urine.
ESTEf3AN& GCNZi\LES' TEXTBOOK OF HISTOLOGY
Fig. XV-1.2.Cell of Proximal Convoluted Tubule.
The electronmicrograph shows the basal lamina
(bl) and basal infoldings of the plasmalemma
(bi) of an epithelial cell that lines a proximal
convoluted tubule. x3,OOO.
The renal tubule consists of several
segments. From the most proximal to the most
distal, they are the proximal convoluted
tubule (PCT), loop of Henle, and distal
convoluted tubule (DCT).
The wall of all the segments of the renal
tubule is made up of a simple epithelium.
However, the shape and surface modifications
~~the epithelial cells vary from segment tq
segment.
Proximal Convoluted Tubule
The proximal convoluted tubule (peT) is
the direct continuation of Bowman's capsule. It
is the longest segment of the renal tubule and is
highly tortuous.
The PC'I's, like the renal corpuscles, are
confined to the cortex. In histologic specimens,
they comprise the majority of the tubules seen
in the cortex.
The PC'I' starts at the urinary pole of the
renal corpuscle. Here, the simple squamou..s
epithelium that comprises the parietal layer
of Bowman's capsule gives way to the simple
cuboidal epithelium of the peT.
In routine histologic preparations, the cells
of the PC'I' are seen to have a centrally located
nucleus and acidophilic cytoplasm due to the
presence of numerous mitochondria. Their
luminal surfaces are provided with thousands
of microvilli (brush border) that, at times,
practically occlude the lumen. Their lateral
borders, on the other hand, are not distinct
because the cell membranes of adjacent cells
interdigitate. Their basal surfaces, meanwhile,
have discernible striations due to infoldings
of the plasmalemma. These basal infoldings
which are loaded with mitochondria increase
the surface area and, therefore, the absorbing
capacity of the cells.
The main function of the PCT is to reabsorb
as much as 70% to 80% of the water and sodium
ions in the glomerular filtrate. This segment of
the renal tubule also reabsorbs other substances,
notably glucose, amino acids, and chloride ions.
The Loop of Henle
The loop ofHenle refersto the section ofthe
renal tubule between ~heproximal convoluted
and distal convoluted tubules. It starts in the
cortex as the continuation of the PCT then
dips into the medulla where it makes a hairpin
turn and then returns to the cortex. It has three
segments: thick descending limb, thin limb, ..
and thick ascending limb.
Thick Descending Limb of the Loop of
Henle (Descending Straight Tubule;
Proximal Straight Tubule)
The thick descending limb is the initial
segment of the loop of Henle. It is a straigh
pipe that is partly in the cortex and partly in the
medulla. It has the same histologic structure
and functions (although it is not as permeable to
ions) asthe PCT and can be regarded asmerelya
part of the PCT that has straightened out.
Thin Limb of the Loop of Henle __
The thin limb of Henle's loop is the
continuation of the thick descending limb. It is
confined to the medulla. It has a much narrower
diameter (15 urn) than the descending thick
limb (60 l-lmon the average).Its wall consists of
a simple squamous epithelium whose cells are
so flat that their nucleus bulges into the lumen
of the tube.
The main function of the thin limb of
Henle's loop, especially in the long-looped
nephrons, is to further concentrate glomerular
filtrate.
Thick Ascending Limb of the Loop of Henle
(Ascending Straight Tubule; Distal Straight
Tubule)
The thick ascending limb of Henle's loop is
the continuation of the thin limb. It starts in the
medulla then heads for the cortex where it seeks
and gets into contact with the vascular pole of
the renal corpuscle of its parent nephron.
Histologically and functionally, the thick
ascending limb of Henle's loop is identical with
the distal convoluted tubule.
Distal Convoluted Tubule
The point of contact of the ascending thick
limb of the loop of Henle with the vascular pole
of the renal corpuscle of its parent nephron
marks the end of the loop of Henle. It is the
beginning of the distal convoluted tifbulc
(DCT), the last segment of the nephron.
Like the renal corpuscles and PCTs, the
DCTs are entirely within the cortex. They are
shorter and less convoluted, but have a bigger
lumen than the PCTs.
In routine histologic preparations, it is
noted that like the PCT, the wall of the DCT is
formed by a single layer of cuboidal epithelial
cells. But in the DCT, the epithelial cells are
shorter and their cell boundaries are distinct.
Furthermore, they do not possess brush borders
and their cytoplasm is less eosinophilic.
The DCT reabsorbs a little amount ofwater
and sodium ions from, and secretes potassium
and hydrogen ions into, the glomerular filtrate.
Classification of Nephrons According to the
Length of their Loop of Henle
N ephrons are classified into two types
based on the length oftheir loop ofHenle: short-
looped and long-looped. The short-looped
nephrons are otherwise known as cortical
URINARY SYSTEM _
nephrons while the long-looped nephrons are
referred to as juxtamedullary nephrons.
Cortical nephrons comprise the great
majority of nephrons in the kidney. Their renal
corpuscles are located in the outer portion of
the cortex. Their loops of Henle which form
part of the medullary rays barely make it to the
medulla. Their loops of Henle consist of a thick
descending limb, a short descending thin limb,
and a thick ascending limb. Their "hairpin loop"
or arch is formed by the initial portion of their
thick ascending limb.
The juxtamedullary nephrons, on the
other hand, comprise about a seventh of
the nephrons in the kidney. Their renal
corpuscles are also in the cortex but near the
corticomedullary junction. Their long Henle's
loops that extend deep into the medulla
consist of a thick descending limb, a long thin
limb that forms the "hairpin loop" or arch,
and a thick ascending limb. Inasmuch as the
"hairpin loop" in these nephrons is formed by
the thin limb of Henle, the thin limb has both
descending and ascending segments.
The EM reveals morphological differences
between the squamous epithelial cells that
Fig. XV-13. Components of the Medullary Ray.
A medullary ray consists of descending thick
limbs (dlt) and ascending thick limbs (atl) of
the loops of Henle, and collecting tubules (ct).
Kidney, H&E x40.
ESTEBAN & CiONZ/\LES' TEXTBOOK OF HISTOLOGY
form the wall of the thin loop of Henle in short-
looped and long-looped nephrons. In short-
looped nephrons, the epithelial cells are of a
single cell type (type I cells). Meanwhile, in the
long-looped nephrons, three cell types different
from type I cells are distinguishable. These cell
types have been labeled tYJ?esII, III, and IV.
The morphological differences among these
cell types (which are beyond the scope of basic
histology textbooks) mainly account for the
functional differences between the long-looped
and short-looped nephrons.
Juxtaglomerular Complex (JG Complex; JG
Apparatus)
At the vascular pole of the renal corpuscle,
there are three groups of atypical cells closely
associated with each other. these cells are
collectively referred to as the juxtaglomerular
complex. These include: 1) JG cells in the
tunica media of the afferent arteriole; 2)
cells that comprise the macula densa in the
distal convoluted tubule (ncr), and 3)
.extraglomerular mesangial cells that occupy
the space between the macula densa and the
afferent arteriole.
Fig. XV-14. Afferent Arteriole. The
electron micrograph shows an afferent arteriol
(aa)with a JG cell (jg) on its wall. x2,500.
 The JG complex helps regulate systemic
blood pressure. In addition, some of its cells
secrete hormones.
JG Cells (Juxtaglomerular Cells)
Just before it enters the vascular pole of
the renal corpuscle, the wall of the afferent
arteriole gets modified. The internal elastic
lamina disappears; the tunica adventitia thins
out; and the smooth muscle cells in the tunica
media are replaced by polyhedral cells called
JG cells. These cells are larger than ordinary
smooth muscle cells.
JG cells have a spherical nucleus and
contain myofilaments and numerous secretory
granules in their cytoplasm. They produce
renin and possibly thrombopoietin. Renin, also
known as an angiotensinogenase, is an enzyme
that participates in the body's renin-angiotensin
system (RAS) that regulates the body's mean
arterial blood pressure.
Macula Densa
In the segment ofthe DCT that isin contact,
with the vascular pole of the renal corpuscle of
its parent nephron, the epithelial cells on the
side of the tubule that abuts on the afferent
arteriole are atypical. They are more crowded
and narrower than those in the other segments
ofthe DCT and rest on avery thin basal lamina.
In routine histologic preparations, their nucleus
appears to be more intensely staining than the
other epithelial cells. These atypical cells are
collectivelyreferred to as the macula densa.
The cells of the macula densa are sensitive
to the sodium ion concentration and water
volume of the fluid in the distal convoluted
tubule. When appropriate, they generate
molecular signals that promote renin secretion
by the JG cells.
The macula densa in the DCT and the
JG cells in the afferent arteriole are in close
proximity with each other. They are separated
only by extraglomerular mesangial cells.
Fig. XV-1S. Macula Densa. The photomicrograph
shows a macula densa (md) that abuts on
an afferent arteriole (aa). The section also
demonstrates a glomerulus (g), Bowman's space
(Bs), parietal layer of Bowman's capsule (pB),
proximal convoluted tubules (pcv), and distal
convoluted tubules (dct). Kidney, H&E x400.
, Extraglomerular Mesangial Cells (Pole-
cushion cells,Lacis cells, Goormaghtigh
cells,Polkissen)
The extraglomerular mesangial cellsform a
conical mass between the macula densa and the
afferent arteriole. In routine histologic sections,
they are seen as flattened and light-staining
cells. Their function has not been definitely
established yet. They are probably involved in
signal transmission between the macula densa
and the glomerular mesangial cells.
Intrarenal Ducts
The intrarenal ducts consist of the
collecting tubules and the papillary ducts (of
Bellini).
A collecting tubule is the continuation of a
DCT. It starts in the cortex as a short segment
that curves towards the medulla (arched
collecting tubule) then straightens (straight
collecting tubule) and heads for the medulla.
URINARY SYSTEM +B
Fig. XV-16. Renal Medulla. The main components
of the renal medulla are segments of the renal
tubules and intrarenal ducts including thin limbs
(tl), descending thick limbs (dt), and ascending
thick limbs (at) of Henle's loop, and collecting
tubules (ct). Kidney, H&E x400

The proximal segments (i.e., segments in

the cortex) of the straight collecting tubules are
the main components of the medullary rays.
Their average initial diameter is 40 ~m.
On reaching the medulla, straight collecting
tubules unite to form larger collecting tubules.
These tubules then merge to form still larger
straight collecting tubules. This convergence of
straight collecting tubules occurs several times
until a large tube (100 to 200 ~m in diameter),
called papillary duct, is formed. the openings of the papillary ducts is called the
area cribrosa.
The straight collecting tubules and papillary
ducts are the main occupants of the renal The lining epithelium of the collecting
pyramids. The only other elements present in tubule is simple cuboidal in its initial segment.
the pyramids are portions of the loops ofHenle, It progressively becomes taller as the tubule
vasa recta, nerves, and minimal amount of becomes bigger. In the papillary duct, the
connective tissue. Each pyramid has about epithelium is tall columnar.
2S papillary ducts whose terminal portions . . In routine histologic preparations, the
come together to form the renal papilla or apex epithelium of the collecting tubule is noted to
of the pyramid, which fits into a minor calyx. consist of cells that have relatively distinct lateral
The region on the renal papilla that-contains borders and faintly eosinophilic cytoplasm that

Fig. XV-17. Papillary

Ducts. Note the simple
columnar epithelium
that lines the wall of the
papillary ducts (pd) in
the photomicrograph.
Kidney, H&E x200.

ESTEBi\\j & GO\.jZALES' TEXTBOOK OF HISTOLOGY

contains a minimal number of organelles. In Histologic Organization of the
the segment of the collecting tubule that is in Urinary Passages and Bladder
the cortex} a second type of epithelial cell} the
intercalated cell} is distinguishable. This cell The wall of the urinary passages and the
is darker-staining than the typical lining cell. urinary bladder has the same basic histologic
structure. It consists of a mucosa}a muscularis
The typical epithelial cells of the collecting
(muscular layer), and an adventitia/serosa.
tubules have been shown to secrete potassium
while the intercalated cells probably playa role
in maintaining the acid-base balance of body
fluids. Thus} the collecting tubules are not mere
conduits for urine.

Interstitial Tissue of the Kidney

The kidney} especially its cortex}
contains very little interstitial tissue.
Nevertheless} some amount of connective
tissue exists in the peritubular and periarterial
spaces. This connective tissue consists of
glycosaminoglycan-rtch ground substance.
Hence} collagenous fibers} capillaries} and
cellular elements that include fibroblasts}
mononuclear cells that probably belong to
the mononuclear phagocyte system} and
fibroblast-like cells-called interstitial cells
(peritubular fibroblasts) -are embedded.
The interstitial cells which vary in shape
possess long processes that are thinner than
those of fibroblasts} and lipid-containing
granules in their cytoplasm. They synthesize
erythropoietin. Some investigators believe
that they also produce prostaglandins. Others
say they secrete a hormone whose effects are
opposite those of renin.

URINARY PASSAGES D
URINARY ,LADDE
From the papillary ducts}urine flows into
the intrarenal urinary passages (i.e., minor
calyces}major calyces}renal pelvis) and then
to the ureter. The ureter conveys the urine
into the urinary bladder where it is stored
temporarily. During micturition} urine is
expelled to the external environment via the
urethra.
Fig. XV-18. Ureter (top). The section illustrates
the histologic layers of the ureter: epithelium (e),
lamina propria (lp), muscularis(m),and adventitia
(a). H&E x1 00.
Fig. XV-19. Transitional Epithelium (above).
Umbrella cells (at arrows) characterize the
transitional epithelium that lines the urinary
bladder and urinary passages. Urinary Bladder,
H&E x400.

URINARY SYSTEM 6ii

Mucosa of the Urinary Passages longitudinally-arranged while those in the
and Bladder outer layer are mostly circularly-arranged.
Many muscle bundles, however, are irregularly-
The mucosa of the urinary passages and arranged and anastomose with each other
urina~y bladder consists of ari epithelium extensively.
that rests on a relatively thin basal lamina and
lamina propria. In the calyces and pelvis, the muscular layer
is thin. In the ureters, it is relatively thicker and
In the ureter, the mucosa forms longitudinal the two layers can be appreciated in routine
folds that impart a stellate appearance to the histologic preparations. In the lower portion
lumen of the organ. In the empty urinary of the ureter, a third muscle layer made up of
bladder, the mucosa forms irregular longitudinal predominantly longitudinally-arranged smooth
folds. muscle cells is present external to the circularly-
The epithelium of the urinary passages and arranged muscle cells.
urinary bladder is transitional. It is essentially In the urinary bladder, the muscular layer
a stratified epithelium where the number is well-developed. It consists of three still
of cell layers increases as one goes from the indistinct but definite layers: two layers of
calyces to the urinary bladder. In the calyces, mostly longitudinally-arranged muscle fiber
the epithelium is f~om 2- to 3-cell thick; in the layers that sandwich a thick layer of circularly-
ureters, it is 4- to S-cell thick; and in the urinary arranged muscle fibers.
bladder, it is up to 8-c~1l thick. Incidentally, At the junction of the bladder and the
transitional epithelium is sometimes referred urethra (i.e., bladder neck) in the male, the
to as urethelium because it is unique to the smooth muscle cells form a definite circular
urinary passages and urinary bladder.
The lamina propria in the calyces and the
pelvis is relatively thin, but in the ureters and the
urinary bladder, it is thick. It consists. of loose
connective tissue that is rich in elastic fibers. It
has a fair supply of MALT and small lymphoid
nodules are occasionally present. It is devoid of
glands except in a limited area near the internal
urethral sphincter located in the neck of the
organ where some mucus-secreting ones are
present.
In the urinary passages and bladder, there is
no distinct submucosa. The connective tissue of
the lamina propria blends with the connective
tissue that encloses the bundles of muscle fibers
in the muscular layer.

Muscular Layer of the Urinary

Passagesand Bladder
In most of the urinary passages, the Fig. XV-20. Urinary Bladder. The low-
magnification photomicrograph shows the
muscular layer consists of two poorly-defined histologic layers of the urinary bladder:
layers of smooth muscle cells that form bundles. transitional epithelium (e), lamina propria (lp),
The muscle cells in the inner layer are mostly very thick muscularis (m), and serosa (5). H&E x40.

ESTEB;\\i & GO\iZ/\L_[S' TEXTBOOK OF HISTOLOGY

collar to comprise the internal urethral
sphincter. In the female, however, the muscle
cells in the bladder neck are arranged obliquely
and longitudinally. Because of this, many
authors say the female does not have a true
internal urethral sphincter.
Adventitia/Serosa of the Urinary
Passages and Bladder
The urinary passages and bladder are
largely retroperitoneal and only the upper part
of bladder is covered by peritoneum. Hence,
the external layer of the walls of the urinary
passages and-most of the bladder are made up
of adventitia that blends with the surrounding
connective tissue. Only the layer in the upper
part of the urinary bladder is made up of serosa.
The adventitia/serosa contains blood vessels,
lymphatic vessels,and nerves.
URETHRA
The urethra is the terminal portion of
the urinary system. In the female, it belongs'
exclusivelyto the urinary system. In the male,
however, it is a tube common to the urinary
and reproductive systems. Aside from serving
as a passageway for urine, it also serves as a
passageway for sperms and the secretions of
the accessory glands ofthe male reproductive
system.
Male Urethra
The male urethra is about 20 em in length
and has three segments. The first segment
(prostatic urethra) traverses the prostat~
gland and is 3 to 4 cm long.The second segment
(membranous urethra) passes through
the sphincter urethrae muscle (external.
urethral sphincter) and is about 1 em long.
The third and longest segment (spongy
urethra, penile urethra, cavernous urethra)
traverses the penis. It is about 15 ern long and
terminates in the external urethral orifice
(meatus).
Prostatic Ureth ra
The prostatic urethra, like the urinary
bladder, is lined with transitional epithelium.
Its lamina propria is a poorly developed loose
connective tissue layer that has many elastic
fibersand veins.It contains afewsmallbranched,
tubuloalveolar, mucus-secreting glands called
urethral glands (of Littre) that empty into the
lumen of the urethra. Incidentally, the urethral
glands (of Littre) are also present in the other
segments of the male urethra.
External to the lamina propria are smooth
muscle fibers which are mostly circularly-
arranged, External to the smooth muscles is
prostatic tissue.
The prostatic urethra also receives the
ducts ofthe prostate gland and the ejaculatory
duct (i.e., the duct that is formed by the union
of the ductus deferens and the duct of the
seminal vesicle).
Membranous Urethra
The membranous urethra differs
histologicallyfrom the prostatic urethra because
its epithelium is pseudostratified columnar or
stratified columnar. The membranous urethra
is also surrounded not by prostatic,tissue, but by
Circularly-arrangedskeletal (voluntary) muscle
fibers that belong to the sphincter urethrae
muscle.
Spongy Urethra (Penile Urethra;
Cavernous Urethra)
The spongy urethra has the same type of
epithelial lining as the membranous urethra,
except in certain areas and where terminally
the epithelium is nonkeratinized stratified
squamous.
The urethral glands (of Littre) in the
lamina propria of the spongy urethra are
decidedly more numerous than those in the
other urethral segments. They empty into
recesses in the mucosa called lacunae of
Morgagni. Incidentally, in the epithelium in
URINARY SYSTEM _

the lacunae ofMorgagni, many of the cells form
intraepithelial glands.
External to the lamina propria is erectile
tissue (see section on penis, Chapter XVI) that
comprises most of the substance of the penis.
The spongy urethra also receives the ducts
of the bulbourethral glands (of Cowper)
which are a pair of small (about 1 em in diameter
each), mucus-secreting glands embedded in the
sphincter urethrae muscle on either side of the
membranous urethra.
Female Urethra
The female urethra is much shorter than
the male urethra. It is only 4 em long. This is
the reason why urinary tract infections which
are usually ascending infections (i.e., the
pathogens enter via the urethral meatus) are
more common in women than in men. The
female urethra is attached to the anterior wall
of the vagina throughout its length. It opens in
ESTE3r\N& C()NZ/\LES' TEXTBOOK OF HISTOLOGY
Fig. XV-21. Spongy Urethra.
The photomicrograph shows the
stratified columnar epithelium (e)
and lamina propria (Ip) that exhibits
a urethral gland of Littre (gL). H&E
x100.
front of the vaginal opening on the vestibule of
the external genitalia.
The mucosa of the female urethra is
thrown into longitudinal folds. Its epithelium
is transitional in most of its length, but becomes
nonkeratinized stratified squamous near the
urethral orifice. Its lamina propria is a loose
connective tissue that contains numerous elastic
elements and many small, mucus-secreting
urethral glands that open into the lumen. The
more external part of the lamina propria, like the
corpus cavernosum of the male penis, contains
a plexus of veins associated with smooth muscle
fibers.
The muscular layer of the female
urethra consists of an internal coat of mostly
longitudinally and obliquely-arranged smooth
muscle cells and an outer coat of circularly-
arranged skeletal (voluntary) muscle fibers that
comprise the sphincter urethrae muscle.
The tunica adventitia consists of a thin layer
ofloose connective tissue.
 apter ~\l1

Male
Reproductive
System

R
eproduction or procreation, the
production of a new individualis arguably
the most imporlant of the biological
processes that humans undertake because it
ensures perpetuation of the species.
Humans procreateby sexual reproduction'
aprocessthat requires the gametes (germ cells)
of the male and the female to unite to form a
zygote'or fertilized ovum. The male gamete
is called sperm cell (spermatozoon) while the
female gamete is called egg cell (ovum).
The gametes are unique among the
numerous cell types in the body in that
they ane haploid (i.e., they possess only 23
chromosomes) while all the others are diploid
(i.e., they contain 46 chromosomes). When a
sperm celland an ovum unite-i-a process called
fertilization-the resulting cell (zygote) is
diploid because it inherits all the chromosomes
in both gametes. Shortly after it has formed, the
zygote undergoes mitotic divisions (cleavage)
and so on becomes an embryo (i.e., the
developing human individual from the time of
implantation to the end of the eighth week after
fertilization). Later, it becomes a fetus (i.e.,the
developing human individual from the end of
the eighth week after fertilization until birth).
The male and female reproductive
systems are responsible for gametogenesis,
the production of male (spermatogenesis) and
femalegametes (oogenesis). They also produce
the hormones that account for the profound
anatomical and physiological differences
between the sexes. In addition, in the male,
the organ system provides a means for the
male gametes to be deposited into the female
genital tract while in the female, it provides the
appropriate milieu for a successful fertilization
and pregnancy.
In males, the reproductive system consists
of a pair of testes, the corresponding duct
system of each testis, a copulatory organ
(penis), and some accessory glands.
DEVELOPMENT OF THE MALE
GAMETES
The earliest recognizable stem cells of the
male and female gametes are calledprimordial
germ cells. They arise from the endoderm of
the yolk sac from the 2nd to the 8th week 0
intrauterine life. Starting on the 4th week of
intrauterine life, these cells begin to migrate
to the developing gonads, undergoing
mitosis along the way. They start to reach the
developing gonads by the end of the 5th week
of intrauterine life. In the developing gonads
(ovaries) of genetic females, these cells soon
differentiate into oogonia, the precursor cells
of ova while in the developing gonads (testes) the male becomes sexually mature, dramatic
of genetic males, they promptly differentiate increase in the production of the male hormone
into spermatogonia, the precursor cells of (testosterone) induces the spermatogonia to
spermatozoa. start to multiply continuously and for some of
I
their progenies to enter th spermatogenic cell
Spermatogenesis cycle (i.e., undergo differentiation.)
.. There are several types of spermatogonia in
Differentiation of the spermatogonia
the testes of the sexually mature male, but the
into sp erm atoz.o a, which occurs in the,
main types are dark type A spermatogonium
seminiferous tubules of the testes, is called
(type Ad spermatogonium), pale type A
spermatogenesis ..It is a process that starts at
spermatogonium (type Ap spermatogonium),
puberty and continues until old age.
and type B spermatogonium.
Spermatogenesis consists of thr e
The type Ad spermatogonium is a relatively
stages) spermatocytogenesis, meiosis,
small cell (about 12 ~m in diameter) that has
and spermiogenesis. Spermatocytogenesis
not entered the spermatogenic cell cycle yet. It
refers to the development of spermatogonia
is so-called because its flattened ovoid nucleus
into primary spermatocytes. Meiosis refers
contains dense chromatin material that makes
to the two successive cell divisions that the
it appear relatively dark in routinely prepar d
primary spermatocytes and their daughter cells
histologic specimens. Type Ad spermatogonia
respectively undergo to give rise to haploid
rarely divide in adults and are considered as
cells called' spermatids. Spermiogenesis
dormant reserve stem cells, but when needed,
refers to the transformation of spermatids into
they can mitose either to renew their numbers
spermatozoa.
or to produce type Ap spermatogonia
. . Type Ap spermatogonia, on the other hand,
primordial germ cell
ale cells that have entered the spermatogenic
cycle (i.e., they have started to differentiate).
type Ad spermatogonium
They divide (mitose) actively either to
renew their numbers or to produce type B
type Ap spermatogonium
spermatogonia.

type B spermatogonium When type Ap spermatogonia undergo

mitosis to produce type B spermatogonia,
the cytokinesis or division of the cytoplasm
primary spermatocyte
that occurs during telophase (the last stage
..... meiosis I
.of mitosis) is not completed such that the
secondary spermatocyte
daughter cells remain connected to each other
..... meiosis II
by cytoplasmic bridges. This incomplete
spermatid cytokinesis also occurs in all subsequent
..... spermiogenesis
cell divisions of these daughter cells and
spermatozoon their progenies. Hence, all the cells in the
spermatogenic series that arise from the same
Fig. XVI-1.Stages of Spermatogenesis type Ap spermatogonium remain connected
to each other (and presumably communicate
-Spermatocytoqenesis with each other) until their connections are
In the sexually immature male, the testes severed shortly before they are released into
contain relatively few spermatogonia and the lumen of the seminiferous tubules as sperm
they divide infrequentl . But at puberty, when cells.

ESTEBAN & GONZi\LES' TEXTBOOK OF HISTOLOGY

 Under the LM, type Ap spermatogonia look
like type Ad spermatogonia except for their paler
nucle s-on account of their lightly-staini g,
finely granular chromatin-that usually
contains a couple of nucleoli that are often
associated with the nuclear membrane. Type
B spermatogonia, on the other hand, resemole
type Ap spermatogoni in appearance except
that their nucleus is more rounded.
A type B spermatogonium undergoes
further mitosis after which all its progenies
increase in size and differentiate into primary
spermatocytes.
The primary spermatocyte is the largest cell
in the spermatogenic cycle (i.e., male germ cell
lineage). In routine histologic preparations, it is
seen as having a large nucleu whose chromatin
material is in the form of fine threads or clump-s,
and abundant cytoplasm.
Meiosis
Meiosis involves two successive cell
divisions (Meiosis I and II), but during this
process, the chromosomes replicate only once'.'
Meiosis I is undertaken by the diploid
primary spermatocyte and results in the
formation of two haploid cells (secondary
spermatocytes). Each secondary spermatocyte
then undergoes meiosis II to produce two still
haploid cells (spermatids) each.
First Meiosis (Meiosis I)
A primary spermatocyte, like all
diploid cells, has 22 pairs of homologous
somatic chromosomes and a pair of male sex
chromosomes (X and Y).
Soon after a primary spermatocyte has
formed and while it is still at interphase
(preleptotene stage), its chromosomes
replicate resulting in the production of two
sister chromatids (as the replicas are called)
per chromosome. Then, it starts meiosis 1.
Meiosis, like mitosis, consists of fou
stages: prophase, metaphase, anaphase, and
telophase.
Prophase of meiosis I consists of five stages
that are distinguishable under the LM: 1)
leptotene stage-during which the chromatin
material starts to condense and form fine threads
within the nucleus; 2) zygotene stage-during
which the chromatin material condenses further
and the homologous and sex chromosomes
approach each other to form 23 pairs (with
each chromosome, because it replicates in the
preleptotene stage, consisting of a pair of sister
chromatids); 3) pachytene stage-during
which a) the chromosomes become thicker byj
condensing even further, rendering the sister
chromatids recognizable as distinct entities,
and, b) corresponding segments of the non-
sister chromatids of each pair of homologous
chromosomes are exchanged, a process called
crossing-over or genetic recombination,
which ensures that the genome of the resulting
offspring is uniqu (note: the sex chromosomes
also exchange genetic material but to a much
lesser extent than the somatic chromosomes);
4) diplotene stage-during which the
homologous and sex chromosomes begin to
separate; and 5) diakinesis-during which
the nuclear membrane disappears and the
homologous and sex chromosomes move a little
further from each other.
Ittakes about 22 days to complete-prophase
of meiosis 1. Following this long prophase
stage" the primary spermatocyte goes through
metaphase, anaphase, and telophase rapidly.
During metaphase, the homologous and
sex pairs of chromosomes align themselves in
parallel fashion at the center of the cel .
At anaphase, one member of each of the
22 pairs of homologous somatic chromosomes;
and the Y chromosome move to one pole of
the primary spermatocyte while the other
member of each of the 22 pairs of homologous
somatic chromosomes and the X chromosome
move to the opposite pole of the cell. Thus,
during anaphase the homologous and sex
pairs of chromosomes part company, but the
sister chromatids of each chromosome stay
togethe.
MALE REPRODUCTIVESYSTEM __
 At telophase, each of the two groups of
chromosomes that have migrated to the opposite
ends of the cell acquires a nuclear envelope.
Also, the cytoplasm of the mother cell is divided
equally between the two daughter cells, but as in
the mitoses of the type B spermatogonium, the
resulting two daughter cells, called secondary
spermatocytes, remain connected to each
other by cytoplasmic bridges. One secondary
spermatocyte has 22 somatic chromosomes
plus the Y chromosome while the other has 22
somatic chromosomes plus the X chromosome,
but all these chromosomes come in duplicate
pairs called chromatids.
A secondary spermatocyte is only about
half the size of a primary spermatocyte.
Second Meiosis (Meiosis II)
As soon as it has formed, the secondary
spermatocyte starts meiosis II, which it
completes in just a matter of hours. Thus, in
routine histologic preparations, secondary
spermatocytes are difficult to find.
In meiosis II, the chromosomes condense
during prophase and move to the equatorial
phase during metaphase. At anaphase, the
sister chromatids of each chromosome are
pulled away from each other. One set of sister
chromatids migrates to one pole ofthe cellwhile
the other set migrates to th~ opposite pole. At
telophase, each set of chromatids acquires a
nuclear envelope and the cytoplasm of the
mother cell is divided equally between the two
daughter cells, but as in the first meiosis, the
resulting daughter cells, called spermatids,
remain connected to each other by cytoplasmic
bridges.
Spermatids contain 23 chromosomes,
but none of them is identical to any of the
chromosomes of their original primary
spermatocyte because of the crossing-over that
occurs during meiosis. In the seminiferous
tubules, the spermatids are located near the
lumen. They are easy to spot in routine LM
preparations because they are smallest and most
numerous of the immature gametes.
ESTES/\N& GONZ!,LES' TEXTBOOK OF HISTOLOGY
Spermiogenesis
During spermiogenesis, the haploid
spermatid which, incidentally, is no longer
capable of cell division, undergoes radical
change in form and becomes a sperm cell
(spermatozoon): its nucleus condenses and
elongates; it forms an acrosome (acrosomal
cap); and it acquires a tail (flagellum).
Spermiogenesis consists of three stages:
Golgi phase, acrosomal phase, and maturation
phase.
In early Golgi phase, the Golgi complex
of the spermatid produces numerous small,
---. .
membrane-bound granules (proacrosomal
granules) that later coalesce to form a Single,
large, membrane-bound structure (acrosomal
vesicle). Meanwhile,. the nucleus starts to
condense and elongate. At the same time, the
centrioles move towards the surface of the cell
that is opposite the aerosomal vesicle. There,
they align with the long axis of the nucleus and
start to elongate to form a tail (flagellum).
" Throughout the Golgi phase, there is
continuous production of proacrosomal
granules by the Golgi complex and fusion of
these granules with the acrosomal vesicle.
Towards the end of the Golgi phase, the
aerosomal vesicle gets attached to one pole of
the nucleus.
During the acrosomal phase, the aerosomal
vesicle transforms into an acrosome-a cap-
like structure that coversa bigpart ofthe nuclear
surface. At the same time, the nucleus elongates
further and its chromatin material condenses
some more while the tail continues to lengthen.
The cytoplasm, on the other hand, migrates,
taking with it all the mitochondria, to surround
the first part of the tail, where it forms a thick
segment called middle piece.
During the maturation phase, the
transformation of the spermatid into a
spermatozoon is completed-residual
cytoplasm is shed and phagocytosed by
the Sertoli cells (see page 247) and the
mitochondria get arranged in a helical manner
around the middle piece forming what is known
as a mitochondrial sheath.
Spermiogenesis ends with spermiation
(i.e., release of the spermatozoon into the
fluid-filled lumen of the seminiferous tubule).
At spermiation, the cytoplasmic bridges that
have connected the progenies of a type Ap
spermatogonium to each other throughout their
development are severed.
When they are released into the lumen
of the seminiferous tubule, the spermatozoa
are anatomically mature, but physiologically
immature. They are not capable of fertilizing
the ovum and are not motile yet. They undergo Fig. XVI-2. Testis and Its Duct System
additional maturation in the ductus epididymis
by a fibrous sheath. The end piece is the short,
where they stay for some time after leaving the
terminal segment of the tail that is not enclosed
seminiferous tubules, but they do not become
by a fibrous sheath.
physiologically mature until after they are
deposited in the female genital tract. The tail has a basic structure that is
identical to that ofa cilium-its core (axoneme)
Ittakes about 64 daysfor the spermatogonia consists of20 microtubules that exhibit the 9+2
to develop into spermatozoa. arrangement (i.e., there is a central pair that is
" surrounded by nine peripheral doublets).
Spermatozoon
The spermatozoon has two parts: head and TESTES
tail. The testes, the organs where
The head of the spermatozoon is made spermatogenesis and synthesis of the male
up of an elongated, highly condensed nucleus hormone (testosterone) occur are a pair of
that is covered on its anterior two-thirds by oval bodies that are lodged in th scrotum, a
the acrosome. The acrosome contains several sac under the penis that is made up of skin ana
hydrolytic enzymes that, during fertilization, subcutaneous tissue. Each testis is 4 to 5 ern
disperse the cells of the corona radiata and long, 3 cm wide and 2 cm thick, and weighs
digest the zona pellucida that surround the about 14g.
ovum. The testis is enclosed by a tough, fibrous
The very long tail of the spermatozoon has capsule, the tunica albuginea, which is made
four segments: neck, middle piece, principal up of dense irregular connective tissue. At
piece, and end piece. The neck or connecting the posterior surface of the testis, the tunica
piece is the segment that is attached to the albugineaisthickened to form the mediastinum
head. Immediately distal to the neck is the testis. It is at the mediastinum testis where
middle piece, which is about 5 ~m in length blood and lymphatic vessels enter and/or exit,
and surrounded by the mitochondrial sheath. and the ductuli efferentes leave,the testis.
Distal to the middle piece is the principal piece. At the mediastinum testis, the tunica
This segment is thinner, but is much longer albuginea sends connective tissue septa
(50 urn) than the middle piece. It is enclosed (septulae testis) into the substance ofthe testis

MALE REPRODUCTIVESYSTEM +£Ii

that divides the organ, although incompletely,
into about 250 pyramidal compartments called
lobules (lobuli testis).

Testicular Lobules (Lobuli Testis)

The testicular lobules, as previously
mentioned, are occupied by highly-coiled
tubes (seminiferous tubules) that are
supported by a stroma consisting of a highly
vascular reticular tissue that fills the spaces
between the seminiferous tubules. Embedded
in this intertubular connective tissue stroma
are eticular cells, mesenchymal cells, and
macrophages. Also scattered either singly
or in clusters in this connective tissue are the
interstitial cells (of Leydig).
Interstitial Cells (of Leydiq)
The interstitial cells (of Leydig) are
large ovoid cells that, in routine histologic
Fig. XVI-3. Architecture of the Testis. The testis
is enveloped by a capsule called tunica albuginea
(ta) which, on the posterior surface of the organ,
is thickened to form the mediastinum testis (mt).
From the mediastinum testis, connective tissue
septa, referred to as septulae testis (st), invade
the organ to divide it into lobules called lobuli
testis (It). Testis, H&E x40.
Fig. XVI-4. Testis. This section of the testis shows
the tunica albuginea (ta) and part of a lobule.
The main occupants of-the testicular lobules are
highly coiled tubes called seminifeorus tubules
(smt). Testis, H&E x100.
preparations, exhibit a round nucleus tha
contains fine chromatin and one or two
peripherally located nucleoli.The cytoplasm of
these cells is copious and eosinophilic. It does
not contain secretory granule because the cells
release their secretions constitutive y, but it
contains lipid droplets and elongate .crystals
(of Reinke). These crystals, which can be I
-----
seen in well-prepared LM specimens, increase
in number with age, but their significance is
unknown.
The interstitial cells (of Leydig) produce
testosterone, the hormone that isresponsiblefor
the secondary sexualcharacteristicsofthe male.
Testosteroneisalsoessentialto the maintenance
of the epithelium of the seminiferous tubules,
the control of spermatogenesis-in the
absence of testosterone, spermatogenesis does
not proceed beyond the meiosisstage-and the'
proper functioning of the accessory glands of
the male reproductive system.
Production oftestosteroneby the interstitial
cells (of Leydig) is regulated primarily by the
luteinizing hormone (LH) that is secreted by
the pituitary gland.

ESTE8AN& GONZ!\LES' TEXTBOOK OF HISTOLOGY


Fig. XVI-S. Interstitial Cells of Leydig. The cells
in the testis that produce testosterone are called
interstitial cells of Leydig '(at arrows). They have membrane. The nucleus has a visible nucleolus
eosinophilic cytoplasm and are located in the
tissue between seminiferous tubules (smt) in the
lobules. Testis, H&Ex100.
Seminiferous Tubules
" smooth endoplasmic reticulum. The cytoplasm
There are up to four seminiferous tubules
in each testicular lobule. They are small (150
to 250 ~m in diameter) but long (30 to 70 cm)
and highly-coiled tubes that are filled with fluid.
They start as blind or as anastomosing tubes at
the base and terminate at the apical portion of
the lobule, which is located in the mediastinum
testis, by becoming continuous with the tubuli
recti.
It is in the seminiferous tubules where the
male gametes (spermatozoa, sperm cells) are
produced. Because the seminiferous tubules are
numerous and long, the number of spermatozoa
produced by the testes is staggering. Each testis
is estimated to produce 94.6 x 106 spermatozoa
per day.
The wall of the seminiferous tubules
consists of an outer fibrous sheath and an
inner epithelium that has a well-developed
basal lamina. The fibrous sheath contains
contractile cells (myoid cells) that resemble
smooth muscle cells and whose contraction
helps propel the spermatozoa into the duct
system. The epithelium, on the other hand, is
a stratified epithelium whose cells consist of
spermatogenic cells (i.e., gametes in various
stages of development) and supporting cells
called Sertoli cells.
Sertoli Cells
Sertoli cells are large, tall cells. They have
broad bases that rest on the basal lamina but
they taper somewhat as they extend into the free
surface of the epithelium. Their apices project
into the lumen of the seminiferous tubule while
their lateral surfaces form folds that partia-lly
enclose the developing male gametes.
Sertoli cells have an elongated nucleus that
is positioned at right angle to the basement
in routine histologic preparations. They are
well-supplied with cytoplasmic organelles-
abundant mitochondria and lysosomes, a well-
developed Golgi complex, and an extensive
of Sertoli cells also exhibits crystal inclusions
that are similar to those found in the interstitial
cells (of Leydig).
Near their bases, Sertoli cells form
tight junctions with each other. These tight
junctions divide the lumen of the seminiferous
tubule into two compartments: basal and
adluminal. The basal compartment is
occupied by the spermatogonia and this is
where spermatocytogenesis takes place.
Once formed, the primary spermatocytes
migrate to the adluminal compartment where
they undergo all the additional development
necessary to become spermatozoa.
The tight junctions the Sertoli cells form
with each other also act as a blood-testes
barrier that presumably prevents the cells
in the adluminal compartment, which are
antigenically different from somatic cells, from
getting into contact with, and being attacked by,
the cells of the immune system.
MALE REPRODUCTIVESYSTEM __
 The other functions of the Sertoli cells
include nutritional support and protection
of the developing gametes; production of the
fluid that fills the lumen of the seminiferous
tubules, which facilitates the transport of the
spermatozoa through the excretory ducts;
phagocytosis of excess cytoplasm that are shed
by spermatids during spermiogenesis; and,
synthesis and secretion of a number of proteins
and the hormone inhibin.
The activity of the Sertoli cells (and in
effect, spermatogenesis) is increased by follide-
stimulating hormone (FSH) that is secreted
by the pituitary _gland. The inhibin that the
Sertoli cells produce, on the other hand, has
a negative feedback effect on the pituitary
gland. It commands the gland to reduce FSH
secretion. The Sertoli cells are also target cells
of testosterone.
Fig. XVI-6. Sertoli Cells and Developing
Germ Cells. Sertoli cells (Se) are broad and
tall cells whose bases rest on the basal lamina
(bl) and whose apices extend into the free
surface of the epithelium of the seminiferous
tubules. Their lateral surfaces have invaginations
where developing gametes including primary
spermatozoa (ps), spermatids (sd), and
spermatozoa (sp) are lodged. Near their bases,
the Sertoli cells form tight junctions (tj) with
neighboring Sertoli cells to effectively divide the
lumen of the seminiferous tubules into a basal
compartment (be) occupied by spermatogonia
(sa) and an adluminal compartment (ac) where
all the other developing gametes are located.
s
EBM---J CiONZ!\LES' TEXTBOOK OF HISTOLOGY
The developing gametes form four to eight
layers of cells on the lateral surfaces of the
Sertoli cells.The spermatogonia, as previously
mentioned, occupy the basal compartment of
the seminiferous tubules. As the progenies ofthe
spermatogonia differentiate, they move inward
towards the lumen of the seminiferous tubule
and get to occupy successively higher folds on
the lateral surfaces of the Sertoli cells. Thus, the
more mature the developing gamete, the closer
it is to the lumen of the seminiferous tubule.
Spermatozoa that are ready for spermiation
occupy the apices of the Sertoli cells.
Blood and Lymphatic Vessels of
the Testes
The left and right testes are supplied with
blood by the left and right testicular arteries,
respectively, that arise from the abdominal
aorta just below the origins ofthe renal arteries.
Before reaching the testis, the testicular
artery divides into several branches. A few of
~~esebranches supply the epididymis; some of
them anastomose with branches of the artery of
the ductus deferens and cremasteric artery;
Fig. XVI-7. Seminiferous Tubule. The
photomicrograph shows a seminiferous tubule
whose wall consists of a fibrous sheath (fs) and
a stratified epithelium made up of Sertoli cells
(Se)and developing gametes in various stages of
development. The stages include spermatogonia
(sa),primary spermatocytes (ps), spermatids (sd),
and spermatozoa (sy). Testis, H&E x400.
 and the rest penetrate the tunica albuginea at
the mediastinum testis and enter the testis.
Some of the arterioles that enter the testis
course towards the rete testis, which they
supply with capillaries. The others take the
opposite direction and follow the connective
tissue elements through the interlobular areas.
From there, their branches enter the lobules
where, in the connective tissue stroma between
the seminiferous tubules, they break up into
intertubular capillaries that anastomose
extensively.
The venules that drain the capillaries
unite to form collecting venules. Some of the Fig. XVI-B. Tubuli Recti. A tubuli recti (tr) is a
collecting venules drain into the veins in the short tubule lined by a simple epithelium that
tunica albuginea, others empty into the venous connects a seminiferous tubule (st) to the rete
plexuses associated with the rete testis, but all testis. Testis, H&E x100.
the veins converge and exit at the mediastinum Intratesticular Genital Ducts
testis.
The intratesticular genital ducts consist
After emerging 'from the testis, the veins
of the tubuli recti, rete testis, and ductuli
form the pampiniform plexus of veins that
efferentes.
surround the ductus deferens (see page 252)
in the spermatic cord (see page 253). The
Tubuli Recti (Straight Tubules)
pampiniform plexus empties its blood into two ..
to three larger veins which later fuse to form a The tubuli recti are the immediate
single testicular vein. continuation of the seminiferous tubules. They
are short, narrow, and straight tubes that are in
The right testicular vein drains into the
the mediastinum testis. Their wall consists of a
inferior vena cava while the left joins the left
simple epithelium that is supported externally
renal vein.
by dense irregular connective tissue.
Lymphatic vessels of the testis start at
In the initial segment of the tubuli recti,
the intertubular (i.e., between seminiferous
the epithelium is composed simply of Sertoli
tubules) areas. They unite with other lymphatic
cells that have no associated gametes. Distally,
vessels at the interlobular connective. tissue.
the Sertoli cells are replaced by columnar or
,.
,.--..
Lymph vesselsfrom the interlobular connective
cuboidal cells that contain numerous microvilli
tissue drain into larger lymphatic vessels in
on their luminal surface. The tubuli recti empty
the capsule, which converge and exit at the
into the rete testis.
mediastinum testis.
Rete Testis
Ducts of the Testis
The rete testis consists of a network of
The sperm cells that are produced in the anastomosing canals that have irregular lumens
seminiferous tubules are conveyed to the within the mediastinum testis. The canals are
outside by a system of excretory ducts. Some of lined by a simple epithelium where the cells
these ducts are within (intratesticular genital are squamous or cuboidal. The cells rest on a
ducts) while some are outside (extratesticular well-developedbasal lamina and in routine light
genital ducts) the testis. microscopic slides, they are darkly-staining.

MALE REPRODUCTIVESYSTEM +JiI

Fig. XVI-9. Rete -Testis. The rete testis refers to
a network of canals in the mediastinum testis
that is lined by a simple squamous or cuboidal
epithelium (e). Testis, H&E x100.
Fig. XVI-10. Ductulus Efferens. Note the
scalloped appearance of the lumen that lines
the wall of the tube. It is shaped as such because
of the way the two types of epithelial cells-one
type is columnar and ciliated while the other is
cuboidal and nonciliated-are arranged.

The epithelium is enveloped externally by the epididymis by their ciliary movement (their
vascular connective tissue that blends with the ciliary
I
beat is towards the epididymis) while
connective tissue of the mediastinum testis. the nonciliated cells absorb, by endocytosis,
some of the fluid that bathe the spermatozoa
Ductuli Efferentes (Efferent Ductu/es) .and which is secreted by the Sertoli cells in the
seminiferous tubules.
The canals of the rete testis drain into
12 to 20 fine tubules (ductuli efferentes) that External to the epithelium, a thin layer
penetrate the tunica albugineaand exitthe testis of circularly-arranged smooth muscle cells
at its posterosuperior surface. Thus, the distal surrounds each ductulus efferens.
segment of each ductulus efferens is actually
extratesticular.
Immediately after leaving the testis, the
ductuli efferentes become highly tortuous as
they travel the short distance to the head .ofthe
epididymis where they merge to form a single
tube, the ductus epididymis.
The wall of the ductuli efferentes consists
of an epithelium that is supported externally
by a thin layer of circularly-arranged smooth
muscle cells. The epithelium has a "scalloped
appearance" in routine LM preparations
because it is made up of two types of cells
(one type is columnar and ciliated while the
other is cuboidal and nonciliated but contains Fig. XVI-11. Ductulus Efferens. The segment
of this duct within the testis is embedded in
microvilli) that form groups that line the duct connective tissue (ct). Its wall contains some
in an irregularly alternating manner. Th-e smooth muscle cells (sm) external to the
ciliated cells help propel the spermatozoa into epithelium (e). Testis, H&E x200.

ESTEBAN& GONZ!\LES' TEXTBOOK OF HISTOLOGY

 Fig. XVI-12. Epididymis, Head. The head of
the epididymis contains the distal ends of the
ductuli efferentes (ef) and the initial segment
of the highly coiled ductus epididymis (ep).
H&E x40.
Fig. XVI-13. Ductus Epididymis.' The wall
of this duct consists of a pseudostratified
epit': elium (e) whose principal cells possess
stereocilia (s) that is supported externally
by smooth muscle cells (mf) and connective
tissue (ct). H&E x400.

Extratesticular Genital Ducts

The extratesticular genital ducts serve as
passagewaysfor the sperm cells from the testis
to the outside. They consist of the ductus
epididymis, ductus deferens (vas deferens),
ejaculatory duct, and urethra.
the blood vessels that supply the ducts and the
The first three ducts are paired (left and' ,
connective tissue that binds the coils __.
6f the
right) while the urethra, which has alreadybeen
ducts.
discussed in the previous chapter, is-unpaired,
The ductus epididymis is a long but highly
Epididymis and Ductus Epididymis , coiled tube. When uncoiled, it is more than 6
meters long, but its diameter is only 1 mm. Its
The epididymis, which should not be wall is formed by a pseudostratified columnar
confused with the ductus epididymis, is a epithelium that is surrounded externally by
C-shaped structure that occupies an area about smooth muscle and connective tissue elements.
7.S em in length on the superior and posterior
surfaces of the testis. It has three parts: a thick The pseudostratified epithelium of the
head that is associated with the superior pole of ductus epididymis is made up of two types of
the testis, a slender body that is associated with cells-principal cells and basal cells.
the posterior surface of the testis: and a short The principal cellsare more numerous than
tail that is associated with the inferior pole of the basal cells. They are tall columnar in the
the testis. segment of the duct that is in the head of the
The head of the epididymis contains the epididymis but progressivelydecrease in height
distal portions of the ductuli efferentes and along the ductus epididymis such that distally,
the initial segment of the highly coiled ductus they are simply cuboidal. Their apical surfaces
epididymis into which the ductuli efferentes are provided with stereocilia.
drain. The body and tail contain the rest of the The principal cells are responsible for
ductus epididymis. Aside from these ducts, absorbing as much as 90% of the fluid that
the other constituents of the epididymis are originates from the seminiferoustubules (which

MA.LEREPRODUCTIVE SYSTEM Wi
is secreted by the Sertoli cells). In addition,
they may have a secretory function because
the complement of cytoplasmic organelles that
they possess (as seen in electron micrographs)
is sug-gestive of such. But the nature of their
secretion, if indeed they have any, is not known
yet.
The basal cells, on the other hand, are
small, rounded, or pyramidal cells that rest on
the basal lamina in between the bases of the
principal cells. Under the LM, they have a pale-
staining cytoplasm and coarse nuclear material.
The basal cells have few cytoplasmic organelles.
Their function is'probably to serve as stem cells
for the principal cells.
The smooth muscle cells that are located
external to the basal lamina of the epithelium
of the ductus epididymis progressively become
more numerous along the duct. In the initial
segment, they form a thin layer and are mostly
circularly-arranged while in the distal segment,
they form three layers: inner and outer layers of
mostly longitudinally-arranged, and a middle
layer of mostly circularly-arranged fibers.
Fig. XVI-14. Spermatic Cord. This structure
is made up of blood and lymphatic vessels,
nerves, and the vas deferens bound together by
connective tissue to form a cord-like structure. In
the photomicrograph, the histologic layers of the
vas deferens can be appreciated. These are the
epithelium (e), lamina propria (lp), inner muscle
layer (iml), middle muscle layer (mml), outer
muscle layer (oml), and adventitia (ad). H&E x40.
Fig. XVI-1S. Vas Deferens. The epithelium of
this duct (e) is pseudostratified columnar with
stereocilia. Deep in the lamina propria (Ip) is a
very thick muscularis layer (m).The most external
layer is an adventitia (a). In the lumen of the duct
is a bunch of spermatozoa (sc). H&E x100.
The ductus epididymis also serves for
accumulation and storage of spermatozoa.
Likewise, it is in this duct where the spermatozoa
become motile.
In humans, the spermatozoa spend 2 to
6 days in the ductus epididymis, but in other
mammals, the transit time of the sperm cells in
the epididymis could be up to 2 weeks. When a
spermatozoon leaves the ductus epididymis, its
tail is fully motile but it still needs to undergo
capacitation before it can fertilize an ovum.
Capacitation, a biochemical process that
involves changes in the cell membrane of
the spermatozoon, occurs within the female
reproductive tract.
Spermatozoa are fast swimmers; when
released in the vaginal canal during sexual
intercourse, they are able to cover the distance
between the vagina and the abdominal cavity in
about 15 minutes.
Ductus Deferens (Vas Deferens)
The ductus deferens is a fibromuscular tube
that is about 45 cm long. It starts at the most
inferior portion of the tail of the epididymis as
the continuation of the ductus epididymis. From

[STEB/\\J & CiO\jZN ..[S' TEXTBOOK OF HISTOLOGY

the tail of the epididymis, it turns upwards along The adventitia is composed of connective
the posterior surface of the testes and enters the tissue that contains the blood and lymphatic
abdominopelvic cavity via the inguinal canal. vessels, and nerves that supply the duct.
Then it crosses the posterior portion of the The distal segment of the ductus deferens
urinary bladder where, after passing the ureter, is dilated and is called the ampulla. In the
it descends towards the prostate gland. ampulla, in contrast to the more proximal
The ductus deferens, from its initial portions of the duct, the mucosal folds are more
segment and until it emerges from the deep numerous, deeper, and more complex. The
inguinal ring to enter the abdominopelvic lining epithelium of the ampulla is sometimes
merely simple columnar, and its muscular layer
cavity is embedded in the spermatic cord.
is much thinner than that in the other segments
The spermatic cord consists of several of the organ.
structures that enter or leave the testis and
which are bundled together by connective Ejaculatory Duct
tissue. Its constituents are the ductus deferens
The ductus deferens terminates by uniting
and its artery and vein, testicular artery,
with the duct of the seminal vesicle to form a
pampiniform plexus of veins, and lymphatic
short tube, about 2 ern in length, the ejaculatory
vessels and nerves of the testes. The structures
duct, which penetrates the substance of the
that comprise the spermatic cord are enveloped
prostate gland to empty into the posterior part
by three layers of tough dense connective tissue of the prostatic urethra.
(internal spermatic fascia, cremasteric fascia,
and external spermatic fascia). The middle of
these layers (i.e., the cremasteric fascia) contains
longitudinally-arranged skeletal muscle fibers
that belong to the cremaster muscle whose
contraction in response to cold brings the testes
closer to the body to prevent damage to the
temperature-sensitive developing gametes.
The wall of the ductus deferens consists of
three histologic layers: mucosa, muscularis,
and adventitia.
The mucosa forms longitudinal folds that
impart a highly irregular shape to the narrow
lumen of the ductus deferens. It is composed of a
pseudostratified columnar (with stereocilia)
epithelium that is similar to that of the ductus
epididymis except that its height is slightly
lower. The lamina propria, which is made up of Fig. XVI-16. Cross Section of the Penis. The
loose connective tissue, has a lot of elastic fibers. organ consists largely of three cavernous
bodies: two corpora cavernosa (cc) and a corpus
External to the mucosa is a very thick spongiosum (cs) whose central area is occupied
muscularis layer composed of smooth muscle by the spongy urethra (u).The cavernous bodies
fibers that are arranged in three distinct layers. are made up of erectile tissue that consists of
blood vessels (bv) embedded in connective
The muscle fibers in the outer and inner layers
tissue (ct). They are encased by tough connective
are longitudinally oriented while those in the tissue called tunica albuginea (ta), external to
middle layer are circularly oriented. which is skin (s).

MALE REPRODUCTIVESYSTEM WI

Fig. XVI-17. Penis. Thisscanning photomicrograph
shows the skin (s) and subcutaneous tissue (ct)
that encase the penis externally. Deep in the
subcutaneous tissue is tunica_albuginea (ta) that
surrounds the two corpora cavernosa (cc) and
the corpus spongiosum (cs)whose central area is
occupied by the spongy urethra. H&E x40.
The mucosa of the ejaculatory duct also
exhibits folds that project into the lumen. Its
lining epithelium is simple columnar.
In the initial segment of the ejaculatory
duct, the mucosa is surrounded externally by a
thin layer of smooth muscle fibers, but within the
prostate, the ejaculatory duct has no muscular
layer-instead, its mucosa is surrounded by
fibromuscular tissue that is part of the stroma
of the prostate gland.
Urethra
In males, the urethra is the last segment
of the duct system of both the reproductive
and urinary systems. Its structure has been
discussed in the chapter on the urinary system.
PENIS
The penis, the male copulatory organ,
consists of three cylindrical masses (cavernous
bodies) that are bound together by connective
tissue and covered externally by skin. Two of
these cavernous bodies, the corpora cavernosa
ESTEB/\\j & GO\iZN_ES TEXTBOOK OF HISTOLOGY
penis, are structurally identical; they occupy
the dorsum of the penis where they lie side by
side. The third, the corpus spongiosum penis,
lies on the median plane, ventral to the corpora
cavernosa penis. Its enlarged distal portion
forms a conical structure, the glans penis,
which is covered by a fold of skin, the prepuce.
Throughout its length, the central portion of
the corpus spongiosum penis is occupied by the
spongy urethra (penile urethra; cavernous
urethra).
A tough layer of dense regular collagenous
connective tissue that is well supplied with
elastic elements, the tunica albuginea, binds
the three cavernous bodies together and forms
a capsule around each one. In the flaccid penis,
the tunica albuginea of the corpora cavernosa
penis is about 2.0 mm thick, but it thins out to
about 0.5 mm when the penis erects fully.
The cavernous bodies are made up of
erectile tissue. This tissue consists of a
labyrinthine system of anastomosing blood
channels that are lined by endothelium and
separated from each other by connective tissue
that contains elastic and smooth muscle fibers.
The vascular channels in erectile tissue
are ordinarily collapsed, but in the presence of
Fig. XVI-18. Erectile Tissue. This type of tissue
which comprises the cavernous bodies in the
penis consists of interconnecting vascular
channels (vc) that are lined by endothelium (e)
and separated from each other by connective
tissue. Penis, H&E x100.
erotic stimuli, they rapidly fill up with blood.
Penile erection occurs when the smooth
muscles in the erectile tissue relax and allow
blood from the two deep central arteries (also
known as the cavernosal arteries) that run down
the length of each cavernous bodies to fill the
vascular channels of the cavernous bodies.
As the corpora cavernosa fill with blood, they
press against the veins in the outside walls of
the corpora cavernosa, compressing them and
stopping the blood from flowing back out of the
penis. As more blood flows in and less flows out,
the penis erects.
External to the tunica albuginea is loose
connective tissue that serves as hypodermis
of the skin that envelops the penis. The
hypodermis is richly supplied with blood Fig. XVI-19. Prostate. The prostate is divided
vessels and it contains some smooth muscle into poorly defined lobules by connective tissue
cells, but is devoid of fat-cells. septa (s).The lobules contain the acini or alveoli
(a) that are lined by simple or psudostratied
The skin that covers the penis has no hair epithelium (e). In some alveoli or acini, lamellated
follicles and contains a limited number of sweat solid concretions called prostatic concretions (pc)
may be seen. H&E x40.
glands.
In the undersurface of the prepuce and in the seminal vesicles account for 60%while that
the proximal portion of the glans penis, there ofthe prostate gland account for 30%of seminal
are sebaceous glands (glands of Tyson) that volume. The bulbourethral glands contribute
are atypical because they are not associated an insignificant amount to semen, only a drop
with hair follicles. In the uncircumcised penis, orso.
the secretion of these glands often accumulates
and forms a cheesy material (smegma) in the Prostate Gland
space between the prepuce and glans penis.
The chestnut-shaped prostate gland
ACCESSORYGLANDS OF THE is the largest of the accessory glands of the
male reproductive system. Its base, which
MALE REPRODUCTIVE SYSTEM
is about 4 cm in diameter, lies immediately
The accessory glands of the male under the apex of the urinary bladder. Its
reproductive system consist of the unpaired vertical diameter is about 3 em while its
prostate gland, and the paired seminal anteroposterior diameter is about 2 cm.
vesicles and bulbourethral glands (glands Its substance is traversed by the prostatic
of Cowper). The combined secretions of these urethra that passes through the center of the
accessoryglands account for most ofthe volume gland. It is also within the gland that the right
of male ejaculate (semen). and left ejaculatory ducts join the urethra.
Semen averages 2 to 5 mL per ejaculation. The stroma of the prostate gland consists
It contains 20 to 250 million spermatozoa per of dense irregular connective tissue that is
mL, but these cells constitute only about lO% richly supplied with smooth muscle fibers-
ofthe volume of the ejaculate. The secretions of the reason why the prostate is often referred to

MALE REPRODUCTIVESYSTEM +Jd


Fig. XVI-20. Prostate. This low-magnification
photomicrograph of the prostate show the
muscle fibers (mf) that are numerous in the septa,
the endothelium (e) that lines the alveoli (a), and
prostatic concretions (pc) that occupy the lumen
of some of the alveoli. H&E x100.
The submucosal glands, on the other hand,
occupy practically the whole inner third of the
prostate gland. Their ducts also open into the
prostatic sinuses.
Meanwhile, the mucosal glands comprise
a small group of glands that occupy the area
that immediately surrounds the urethra. Their
ducts open directly on the luminal surface ofthe
prostatic urethra.
The epithelium that lines the alveoli of the
prostate gland is often simple columnar but in
some areas it is sometimes simple squamous,
simple cuboidal, or pseudo stratified columnar.
In some of the alveoli, the lumen contains
lamellated solid concretions (prostatic
concretions; corpora amylacea) that are of
variable size, some reaching 2 mm in diameter.
The prostatic concretions are composed of
glycoproteins and other substances that are
deposited around cell fragments. They increase
in size and number, and calcify,with age.

as a fibromuscular organ. The stroma forms a The secretion of the prostate gland is
an alkaline fluid that contains enzymes,
capsule that envelops the gland. The capsule
fibrinolysin, prostaglandins, and a compound
gives off septa that incompletely subdivide the
with antibiotic properties.
prostate gland into about 50 poorly defined
lobules. The blood vessels and nerve's of the
gland ramify in the septa. From the septa, Seminal Vesicles
connective tissue elements and smooth muscle
The seminal vesicles are a pair (left and
cells invade the lobules and create a supporting
right) of sac-like structures that are about 5 em
framework around and between the alveoli.
The parenchyma of the prostate gland
consists of three groups of compound
tubuloalveolar glands that are arranged
concentrically around the urethra: 1) main
prostatic glands; 2) submucosal glands; and,
3) mucosal glands. The ducts of the three
groups of glands all open into the prostatic
urethra.
The main prostatic glands comprise the
bulk, and occupy the peripheral two-thirds of
the prostate gland. Their secretion is collected
by about 20 ducts that open independently of
each other into the prostatic sinuses on the sides Fig. XVI-21. Seminal Vesicle. The illustration
of the urethral crest on the posterior aspect of shows the histologic layers of the seminal vesicle.
the prostatic urethra. Note the intricate folds that the mucosa forms.

ESTESf.\hl & COhlZJ\LES' TEXTBOOK OF HISTOLOGY

long each. They lie between the fundus of the
urinary bladder and the rectum immediately
above the prostate gland on either side of
midline.
Each seminal vesicle consists of a narrow
and long (10 to IS cm) tube whose numerous
<toils are bound together by connective tissue,
which externally, also forms a capsule for the
organ externally.
The wall of the tube has three histologic
layers: mucosa, muscularis, and adventitia.
The mucosa forms numerous intricately
branched folds that give the lumen the
appearance of a maze. Its lining epithelium
is nonciliated pseudostratified columnar Fig. XVI-22. Seminal Vesicle. The scanning
photomicrograph shows several coils of the
(or cuboidal). The cells comprising the
seminal vesicle in cross section. In each coil, the
epithelium are mostly columnar or cuboidal mucosa (rna)forms complex anastomosing folds
cells that contain lipochrome pigments in their that give the lumen (I) a maze-like appearance.
cytoplasm. In between the bases of these cells External to the mucosa are a muscularis layer
are a few small, round cells which are likely stem (mu) and an adventitia (a)which is often common
to adjoining coils. H&E x40.
cells.

Fig. XVI-23. Wall of the

Seminal Vesicle. This high
magnification photomicrograph
shows the epithelium (e) of
the seminal vesicle and the
underlying lamina propria (lp) in
greater detail. The epithelium
is pseudostratified cuboidal
(as in this section) or columnar.
In between the cuboidal or
columnar principal cells are a
few small cells that are probably
stem cells. Muscle fibers (mf) in
the muscularis layer can also be
identified in the section. H&E
x400.

MALE REPRODUCTIVESYSTEM +a
 The muscularis of the seminal vesicle
consists of a thin inner layer of circularly-
arranged and a thicker outer layer of
longitudinally-arranged smooth muscle fibers.
The adventitia is connective tissue with
elastic elements.
The epithelial cells of the seminal vesicle
produces a secretion that contains fructose (an
important source of energy for the spermatozoa),
prostaglandins, and fibrinogen.
Bulbourethral Glands (of Cowper)
The bulbourethral glands are a pair (left and
right) of pea-sized (1.0 cm diameter), yellowish
organs that are embedded in the sphincter
urethrae muscle where they lie behind and
lateral to the membranous urethra.
Each bulbourethral. glana is a compound
tubuloalveolar mucous gland that is enclosed
by a thin connective tissue capsule that contains
collagenous and elastic fibers, and some smooth
and striated muscle cells. Connective tissue
septa from the capsule divide the gland into
ESTEBAN & CONZALES' TEXTBOOK OF HISTOLOGY
small lobules that contain the alveoli and ducts
of the gland.
The alveoli are lined by a simple cuboidal
epithelium. The nucleus of the epithelial cells is
typically displaced towards the base of the cell
because of the presence of numerous secretory
granules in the cytoplasm.
The alveoli drain into ducts that are lined
by simple tall cuboidal or columnar epithelium
that contains patches of mucus-secreting cells.
A confluence of the ducts ultimately gives rise
to the main excretory duct that is lined by a
pseudostratified columnar epithelium that, as
in the smaller ducts, contains patches of mucus
cells.
The main excretory duct is relatively long
(2.5 to 3.0 em), It opens into the proximal part
of the spongy urethra.
The secretion of the bulbourethral
glands consists of a clear, viscous fluid that is
discharged shortly before ejaculation. It helps
to lubricate the urethra for easier passage of the
spermatozoa.
Female
Reproductive
System
he organs that comprise the female

T
reproductive system are grouped into the
internal and external genitalia (vulva).
Internal genitalia refers to the ovaries,
oviducts, uterus, and vagina while external
genitalia refers to the clitoris, labia majora
and minora, and some glands whose ducts open
into the vestibule (i.e., the space into which the
orifices of the urethra and vagina open). "<, '.
The mammary gland, although a
modified sweat gland, is better considered part
of the female reproductive system because of
its function, so it shall likewise be discussed
in this chapter. Another added material in this
chapter is a discussion on implantation and
placentation.
MENS UALCYCLE
The menstrual cycle varies in length from
individual to individual and even in the same
woman from cycle to cycle, but the average
is 28 days. Its most notable manifestation
is menstruation or menstrual flow (i.e.,
vaginal bleeding that lasts for three to five
days). It is customary to refer to the first day of
menstruation as the first day of the menstrual
cycle simply because this is the day in the cycle
that is easiest to ascertain.
The reproductive life of females is
considerably shorter than that of males. It starts
at puberty and is heralded by menarche (first
menstruation), which usually occurs when a
girl is between 11 to 14years of age, and ends at
menopause (cessation of menstruation), which,
on the average, occurs at age 52. From menarche
to menopause, the menstrual cycle is repeated
over and over, interrupted only by pregnancy.

In sexually mature non-pregnant

women, the organs of the reproductive
system, especially the ovaries and the -uterus,
undergo cyclical anatomical and physiological
changes in response to a complex system of
hormonal and neural stimuli. These cyclical ... meiosisll
changes comprise the menstrual cycle.
They are designed to ensure that an ovum .... meiosis I
(female gamete) is periodically available for
fertilization, and that if fertilization occurs, it
results in a successful implantation (see page Fig. XVI-1.Developmental Stages ofthe
277) and pregnancy. Female Gamete
DEVELOPMENT OF THE
FEMALE GAMETE
As discussed in the previous chapter, the
earliest recognizable stem cells of both male
and female gametes are called p.rimordial germ
cells. They arise from the endoderm of the yolk
sac between the second and eighth weeks of
intrauterine life (IUL). Starting on the fourth
week ofIUL, these cells begin to migrate to the
developing gonads, undergoing mitosis along
the way.'
In the female, the primordial germ cells
start to reach the developing ovaries by the end
of the fifth week oflUL where they differentiate'
into the precursor cells (oogonia) of the female
gametes or ova.
In the developing ovaries, oogonia undergo
further mitosis before the daughter cells start
the process of oogenesis that will transform
them into ova.
Oogenesis
Differentiation of the oogonia into ova in
the ovaries is referred to as oogenesis.
The initial stage of oogenesis involves
differentiation of oogonia into bigger cells called
primary oocytes that are no longer capable of
mitosis. Oogonia start to transform into primary
oocytes at the end of the third month ofIUL. By
the seventh month ofIUL, this initial process is
completed, which means all the oogonia in the
ovaries of a female fetus have transformed into
primary oocytes. ( cytoplasm. The ovum is capable of further
Developing female gametes, like developing
male gametes, need to undergo two successive
meiotic divisions (meiosis I & II). It is the
primary oocytes that go through meiosis 1.
In fact, as soon as they have formed, primary
oocytes, like the primary spermatocytes in the
male, replicate their chromosomes, then start
the first meiotic division.
Meiosis I in oogenesis is essentially similar
to meiosis I in spermatogenesis, except that
in oogenesis, the process is suspended in
ESTEBr\N& GOND\LES' TEXTBOOK OF HISTOLOGY
the: diplotene stage of prophase and is not
completed by any of the cells until the female has
reached sexual maturity. The primary oocytes
therefore go into a meiotic resting period called
dictyotene phase that lasts for many years.,The
other main difference of meiosis I between the
sexes is that-as will be elaborated on a few
paragraphs from now-the resulting daughter
cells in oogenesis are not identical and are not
bound to each other by cytoplasmic bridges:
At birth, a female has about two million,
primary oocytes in her ovaries..All these oocytes
are in dictyotene phase. Many of these cells,
will degenerate and die even before the female
reaches sexual maturity. Since primary oocytes
are no longer capable of mitosis, their number
cannot be renewed. Consequently, at puberty,
the number of primary oocytes a female has in
her ovaries has dwindled to only about 400,000.
As a rule, starting at puberty and until
menopause, one-and only one-primary
oocyte is allowed to resume and complete
meiosis I every menstrual cycle. The completion
of the first meiotic division in the chosen oocyte
occurs a few hours before ovulation (see page
265).
Completion of meiosis I produces two
haploid daughter cells: However, practically all
the cytoplasm of the mother cell goes to only one
daughter cell, the secondary oocyte or ovum.
The other daughter cell, the first polar body,
ebds up with nuclear material that is identical to
that of the ovum but without or with minimal
development while the first polar body is not,
and is later extruded, though in some lower
forms of animals, the first polar body has been
observed to undergo meiosis II.
Immediately after it has formed, the ovum
starts meiosis II, which is also similar to meiosis
II of spermatogenesis, except that in oogenesis,
the process is suspended at the metaphase stage
and resumes only if the cell is fertilized.
Ifit is not fertilized by a spermatozoon, the
ovum undergoes no further development and
\ \
 germinal epithelium because early histologists
thought that it is the source of the female germ
cells. Beneath the epithelium is a layer of
Fallopian
tube
dense irregular connective tissue, the tunica
albuginea, which forms a capsule, albeit
indistinct, for the organ.
ovary
The substance of the ovary consists of
a thick peripheral zone called cortex that
surrounds a very vascular inner zone, the
medulla.

Fig. XVII-2. Organs of the Female Internal The cortex contains numerous ovarian
Genitalia follicles that are supported by a stroma of
collagenous connective tissue while the medulla
degenerates. If it is fertilized, it hastily completes
consists ofloose connective tissue that is richly
meiosis IIto produce two very unequal daughter
supplied with blood vessels. An ovarian follicle
cells called second polar body and fertilized
consists of a developing female gamete and the
ovum (zygote).
cells and other tissue elements that encase it.
The second polar body, like the first
polar body, receives nuelear material from its Ovarian Cycle
mother cell but notmuch else. Like the first
polar body, it is later extruded. In contrast, the Ovarian cycle refers to the structural and
zygote receives practically all the cytoplasm physiological changes that some ovarian follicles
of its mother cell. In addition, it is diploid undergo during the menstrual cycle in response
because its nucleus contains not only 23 but 46. to the gonadal hormones (FSH and LH) from
chromosomes since it was formed by the fusion the pituitary gland.
of the male pronucleus (i.e., nuclear material of The ovarian cycle has two phases,
the spermatozoon that fertilized the ovum) and follicular and luteal. The first two weeks of
the female pronucleus (i.e., nuclear material of the cycle, which ends with ovulation, comprise
the ovum). the follicular phase. It is governed by FSH. The
last two weeks, which starts with ovulation,
OVARY
The ovaries are a pair (left and right) of
slightly flattened, ovoid organs, each of which
is about 3 cm x 1.5 cm x 1 ern in size. They are
the sites of oogenesis and they produce a few
hormones.
Each ovary is enveloped by a simple
squamous or cuboidal epithelium called

Fig. XVII-3. Ovary. The parenchyma of the

ovary consists of two regions: a peripherally-
located cortex (c) that surrounds a centrally-
located medulla (m). The cortex contains the
ovarian follicles. In the photomicrograph, the
follicles are in various stages of development:
primordial follicles (plf), primary follicles (pyf),
and secondary follicles (sf). H&E x40.

FEMALE REPRODUCTIVESYSTEM ..
 one l<lyer
~ follicular cells

ffO-'"- ovum
severall"'y'e(~of
follicular ceJl~

prImordial folHcl·(,

fJtimary follicle

Gr.lafi.ln futlid('

Fig. XVII-4. Developmental Stages of the Ovarian Follicle

comprise the luteal phase. It is governed by LH.
During the follicular phase, several ovarian
follicles undergo varying levels of development
while during the luteal phase, the corpus
luteum (see page 266) develops and becomes
functional.
Ovarian Follicle
Three types of ovarian follicles may be seen
in the cortex of the ovaries of sexually mature
females: primordial, primary, and secondary.
follicular epithelial cells) that rest on a thin
basal lamina.
A primary oocyte, the granulosa cells
that surround it and the basal lamina on which
the granulosa cells rest comprise a primordial
follicle. The primary oocyte in a primordial
follicle is about 15 to 30 ~m in diameter while
the whole follicle is about 40 ~m. The primary
oocyte has a large, eccentrically placed,
vesicular nucleus with a large nucleolus. It also
I

Primordial Follicle
As mentioned in the section on oogenesis,
shortly after the oogonia have reached the
developing ovaries during early embryonic life,
they transform into primary oocytes. In the
ovaries, all the primary oocytes get enveloped
by a single layer of flattened epithelial cells
(granulosa cells, squamous epithelial cells,

Fig. XVII-S. Pr irn o d ia l Follicles. The

photomicrograph shows a cluster of primordial
follicles (at unlabeled arrows) in the cortex, a
short distance from the germinal epithelium
(ge). The follicles are made up of an oocyte (0)
that exhibits a prominent nucleus (nu) and a
single layer of flattened granulosa cells (gc) that
surrounds the oocyte. Ovary, H&E x400.

ESTEBAN& GONZALES' TEXTBOOK OF HISTOLOGY

Fig. XVII-6. Primary Ovarian Follicle, Early.
A primary ovarian follicle in its early stage of
development is shown in the section. The
granulosa cells (gc) form a simple columnar
epithelium around the oocyte (0) which has
a centrally-located nucleus (nu). Between the
oocyte and the granulosa cells is a deeply staining
membrane, the zona pellucid (zp). External to the
granulosa cell layer, the cells have differentiated
to form a theca folliculi (tf). The photomicrograph
also shows some primordial ovarian follicles (plf).
Ovary, H&E x400.
Fig. XVII-7. Primary Ovarian Follicle. In this
section, the granulosa cells (gc) around the
oocyte-which still displays a centrally located
nucleus (no)-already form a stratified epithelium
while the theca folliculi (tf) is much thicker than in
the previous figure. Ovary, H&E x400.
A primordial follicle that is destined to
produce an ovum becomes initially)a primary
follicle) and later)a secondary follicle.

has a prominent Golgi apparatus) numerous Primary Follicle

mitochondria) ribosomes) and endoplasmic At the start of an ovarian cycle) several
reticulum. primordial follicles (up to 15) begin
Granulosa cells) on the other hand) development)which is heralded by an increase
are flattened cells that resemble ordinary in size of the flattened granulosa cells. The
connective tissue cells. They are supporting granulosa cells initially become cuboidal-
cells that provide the developing gamete with at which point the follicle is already termed a
nutrients and oxygen. primary follicle-then columnar) and finally)
At birth) the number of primordial follicles as a consequence of repeated mitosis) they
in the two ovaries) which is obviously equal stratify. When there are already several layers
to the number of viable oocytes, is about two ofgranulosa cells)fluid (follicular fluid; liquor
million. Early in life) many of these follicles folltcult), whose exact origin is still unclear)
and the oocytes they contain degenerate; starts to appear between the cells.
many others undergo some amount of Meanwhile) the primary oocyte in the
development) but these too) will not develop follicle gradually increases in size. When
fully and ultimately degenerate. It is only at the oocyte is already about twice its original
puberty) when the number of ovarian follicles diameter) a thick) deeply-staining)glycoprotein
has already dwindled down to about 400)000) membrane-the zona pellucida-develops
that the ovaries become fully functional and a around it. This membrane is probably a joint
few follicles are given the opportunity to fully product of the oocyte and the granulosa cells.
develop. It is through it that the granulosa cells provide

FEMALE REPRODUCTIVESYSTEM ..
nourishment for the oocyte. In electro-
micrographs, microvilli from the oocyte are
seen to extend into the zona pellucida. These
microvilli are in contact, via gap junctions, with
the cytoplasmic processes of the innermost layer
of granulosa cells.
While the oocyte and the granulosa cells
are undergoing the changes described above, the
stroma that immediately surrounds the ovarian
follicle becomes organized to form a sheath, the
theca folliculi, that envelops and becomes part
of the follicle.
The theca folliculi later differentiates into
two layers: an inner cellular and vascular layer
called theca interna, and an outer fibrous layer
called theca externa.
The theca intern a is richly supplied with
capillaries. Its cells are. large and loaded with
lipid. Under the influence of LH, they secrete
androstenedione (a precursor of testosterone),
which seeps into the granulosa cell area where it
is transformed into estrogen through the help
of aromatase, an enzyme that is activated by
the granulosa cells under the influence ofFSH.
Then, estrogen diffuses into the capillaries of
the theca interna to be carried by blood to its
target organs, i.e., uterus and the other organs
of the female reproductive system.
The theca externa, on the other hand, is
essentially a connective tissue layer that can be
regarded as a capsule that envelops and separates
the follicle from the ovarian stroma.
Secondary Follicle (Antral Follicle)
As the liquor folliculi in the primary follicle
increases in amount, the irregular spaces
between the granulosa cells, which contain
the liquor folliculi, become confluent to form
a single crescentic, fluid-filled cavity called
antrum. Once an antrum has developed, the
ovarian follicle has transformed into an antral
follicle or secondary follicle.
A typical secondary follicle is oval in
shape. It has a stratified epithelium consisting
of granulosa cells that displays a thickening
(cumulus oophorus) on one pole. The cumulus
oophorus protrudes into the fluid-filled antrum
and its central area is occupied by the oocyte,
which incidentally stays as a primary oocyte
until shortly before ovulation.
Graafian Follicle (Mature Follicle)
As ovulation approaches, the connection
of the ovum and the granulosa cells that abut
on the zona pellucida with the rest of the

Fig. XVII-B. Secondary Ovarian

Follicle. An ovarian follicle with
an antrum (a) that is filled with
liquor folliculi is referred to as a
secondary follicle. At one pole
of the follicle, note the cumulus
oophorus (co) which consists of
granulosa cells. In the center
of the cumulus oophorus is the
oocyte (0) with its distinctive
nucleus (no). The nucleus is
separated from the granulosa
cells by the zona pellucid (zp).
External to the granulose cell
layer, the two layers of the
theca folliculi-theca interna (ti)
and theca externa (te)-can be
distinguished. Ovary, H&E x100.

ESTEBAN & GONZALES TEXTBOOK OF HISTOLOGY

granulosa cells in the cumulus oophorus is
loosened by the development of new liquor-
filled intercellular spaces.
At about the time that the ovarian follicle
has matured, a sudden surge in LH secretion
by the pituitary gland occurs. This LH surge
causes the primary oocyte in the secondary
follicle to resume and complete its first meiotic
division. As discussed in the section on
oogenesis, completion of meiosis I produces two
haploid daughter cells: the secondary oocyte or
ovum, which practically gets all the cytoplasm
of the mother cell; and the first polar body,
which has nuclear material but with little or no
cytoplasm. The secondary oocyte immediately
starts meiosis II, but this process is suspended
at the metaphase stage and resumes only if
the cell is fertilized. The first polar body, on
the other hand, is tempo-rarily consigned to
the perivitelline space (Le., space that forms
between the cell membrane of the secondary
oocyte and the zona pellucida) prior to its
extrusion.
, .for it to travel from the abdominal cavity into
A fully developed or mature secondary
ovarian follicle is called a Graafian follicle.
It is the final stage in the differentiation of an
ovarian follicle, one that is ready to expel its
ovum or secondary oocyte. It is a big structure
that reaches 1.5 to 2.5 cm in diameter and
it often occupies the whole thickness of the
ovarian cortex and may even bulge out of the
free surface of the organ.
It takes about 14 days for an ovarian follicle
to mature. Normally, as previously mentioned, at
the start of every ovarian cycle, several follicles
in both ovaries begin to develop in response
to FSH (actually the first few days of follicular
growth is independent of FSH), but only one,
the dominant follicle,reaches maturity. All the
others degenerate at various stages. It is not clear
what makes the dominant follicle dominant, but
one plausible explanation is that it elaborates a
protein called follicular regulatory protein
that prevents the other follicles from getting
stimulated by FSH.
Ovulation
The LH surge that triggers completion of
meiosis I by the primary oocyte also causes
the Graafian follicle to burst a few hours later
and release its ovum into the abdominal cavity.
This discharge of the ovum into the abdominal
cavity is what is referred to as ovulation. From
the abdominal cavity, and aided by the ciliary
action of the oviductal mucosa, the ovum finds
its way into an oviduct.
Ovulation occurs on or about the 14th day
of an ideal 28-day ovarian cycle.
When the ovum is released by the ovary,
it carries with it the zona pellucida and one
to several layers of granulosa cells that cling to
the zona pellucida. These granulosa cells are
collectively referred to as corona radiata.
Fertilization
An ovum is viable only for about 24 hours
after being ovulated, but it takes 3 to 4 days
the uterus via an oviduct. Hence, fertilization
(i.e., the union of the male and female
gametes), if it has to occur, has to take place in
the abdominal cavity or the initial part of the
oviduct. In fact, it usually occurs in the distal
third of the oviduct.
Spermatozoa that are deposited in the
vagina are still incapable of fertilizing an ovum.
They still need to undergo capacitation within
the female genital tract. Capacitation is a
biochemical process that involves changes on
the surface of the spermatozoa. It is completed
by the spermatozoa on a staggered basis-
some soon after getting deposited in the female
genital tract while others as long as 5 to 6 hours
later. This is evidently to keep the chance
for fertilization alive over a relatively long
time inasmuch as the exact moment that the
oocyte and spermatozoon will meet cannot be
determined ahead of time.
During fertilization, the acrosome of the
capacitated spermatozoa breaks down-by
FEMALE REPRODUCTIVESYSTEM _
Fig. XVII-9. Fertilization. The series of diagrams illustrates the union of the male and female
gametes that usually occurs in the distal third of the oviduct. Sperm cells approach the oocyte (a) in
the metaphase stage of meiosis II. Once a sperm cell has penetrated the zona pellucida, it becomes
impermeable to the other sperm cells. The entry of the sperm cell triggers the completion of meiosis
II in the oocyte. Then the male pronucleus approaches and unites with the female pronucleus (b, c,
and d) to form a zygote (e).

a process called aero somal reaction-and Corpus Luteum

releasesits hydrolytic enzymes. These enzymes
disperse the cells of the corona radiata by
depolymerizing their intercellular s1;lbstance.
They likewise enable a sperm cell to digest and
penetrate the zona pellucida.
Once a sperm cell has penetrated the zona
pellucida, the zona becomes impermeable to
any other sperm cell.This ensures that only one
sperm cell fertilizes an ovum.
As discussed in the section on oogenesis,
entry of the spermatozoon into an ovum
triggers the completion of meiosis II by the
secondary oocyte that results in the formation
of a diploid fertilized ovum or zygote and a
second polar body, which, like the first polar
body, is consigned to the perivitelline space
prior to extrusion.
Aside from restoring the normal (diploid)
number of chromosomes in the zygote,
fertilization also determines the sex ofthe fetus
and initiates cleavage (i.e., mitotic division of
the zygote).
"
During ovulation, the ovum and liquor
folliculi spill into the abdominal cavity.
Consequently, the wall of the ovarian follicle
collapses. The granulosa cell layer and theca
interna are thrown into folds and their cells,
under the influence of LH, undergo certain
changes-they enlarge, become plump, pale-
staining and polygonal, and they accumulate
lipids. Henceforth, they are called lutein cells
and what used to be the ovarian follicle is now
the corpus luteum.
The cells of the corpus luteum that are
derived from the granulosa layer are known
as granulosa lutein cells, while those derived
from the theca interna are called theca lutein
cells. The granulosa lutein cells comprise about
80% of the substance of the corpus luteum.
They are lighter-staining and larger than the
theca lutein cells (30 ~m to SO ~m vs. 15 ~m, in
diameter).
The two cell types of the corpus luteum
also differ functionally. The granulosa lutein

ESTEBAN & CiCP--JD\LES'TEXTBOOK OF HISTOLOGY

Fig. XVII-10. Corpus Luteum. The scanning Fig. XVII-11. Corpus Luteum. This high
photomicrograph shows a corpus luteum (cl). magnification photomicrograph shows the pale-
Ovary, I-' ~ '- x40. staining, plump, and lipid-loaded lutein cells.
Ovary, H&E x400 .

.cells produce, and are the main source of pituitary gland, the corpus luteum becomes
progesterone, a hor?IoIie that is necessary considerably bigger and remains functional for
for preparing the mucosa of the uterus or about six more months and becomes known
endometrium (see page 270) for reception of as corpus luteum of pregnancy. This is
the conceptus (i.e., the product of conception because another hormone, human chorionic
after fertilization). It is also necessary for the gonadotropin (heG), which is secreted initially
conduct of a succes~ful pregnancy. The theca' . by the embryo and later by the developing
lutein cells, on the other hand, like the theca placenta, takes over from LH in maintaining
interna cells from which they aro~e, secrete and further developing the corpus luteum.
testosterone precursors, which seep into In addition to estrogen and progesterone,
the granulosa lutein cell area where they are the corpus luteum of pregnancy also elaborates
transformed into estrogen. In addition, the another hormone, relaxin. Relaxin is most
theca lutein cells also secrete some amount likely also secreted by the granulosa lutein cells.
of progesterone. The amount of estrogen It helps maintain pregnancy by inhibiting the
produced by the corpus luteum during the contractions of the myometrial muscle fibers. It
second half of the menstrual cycle is almost also aids during delivery by relaxing the pelvic
equal to the amount that is produced by the ligaments and cervix.
ovarian follicle during the first half of the cycle.
Towards the end of the menstrual cycle, Corpus Albicans
LH levels decrease markedly and unless
When the blood level of the hormone that
implantation occurs, the corpus luteum, which
maintains the corpus luteum (of menstruation
is maintained by LH, degenerates. Hence,
or of pregnancy) drops, the lutein cells
unless a particular menstrual cycle results in
.become loaded with lipid and degenerate. In
a pregnancy, the corpus luteum lasts for only
the succeeding months, the corpus luteum is
about 10to 14days and is called corpus luteum
reduced to a white scar in the ovarian cortex
of menstruation.
called corpus albicans. Over aperiod ofmonths
However, if implantation and pregnancy or years, the corpus albicans gradually involutes
occur, despite waning LH secretion by the until it finally disappears.

FEMALE REPRODUCTIVESYSTEM 'fB


Fig. XVII-12. Corpus Albicans. The corpus
albicans (ca) consists of scar tissue. Ovary, H&E
x100.
Atresia of Follicles
More than 99.9% 'of ovarian follicles do
not reach maturity. They instead undergo
degenerative processes at various stages of
development. Out of the approximately two
million follicles a woman has at birth, she will
ovulate less than 500 in her lifetime.
The process of degeneration of ovarian
follicles is referred to as atresia. It occurs
continuously throughout a woman's lifetime.
Atresia can occur at any stage in the
development of a follicle. In a primary follicle,
atresia is first noted in the oocyte-the cell
shrinks. This is followed by degeneration of the
granulosa cells.
In small secondary follicles, usually the first
sign of abnormality is the eccentric location of
the egg nucleus. However, in bigger secondary
follicles, the process is appreciated in the wall
Q{ the, follicle before changes in the oocyte are
noted. Initially, connective tissue strands invade
the spaces between granulosa cells; as a result,
the granulosa cells get loosened and fall into
the follicular cavity (antrum). Meanwhile, the
cells of the theca interna become bigger while
the basal lamina between the theca interna and
the granulosa cells thickens, and is sometimes
ESTES,\\ & GO\Zl\L.ES' TEXTBOOK OF HISTOLOGY
referred to as glassy membrane. Thereafter,
the oocyte slowly deteriorates and after a while,
the only telltale sign that an oocyte was once
present in the area is a collapsed zona pellucida.
Ultimately, the glassy membrane as
well as the zona pellucida degenerate and
disappear, but while they persist, they serve to
distinguish atretic follicles from similar-looking
degenerating corpora lutea under the light
microscope.
Interstitial Gland of the Ovary
There are epithelioid cells in the stroma
of the ovary that cytologically resemble theca
lutein cells. These cells, which gather in clusters,
are numerous during the first few years of life
but are hardly seen in the adult ovary. They
evidently secrete estrogen and are collectively
referred to as interstitial gland of the ovary.
OVIDUCTS (UTERINE TUBES;
FALLOPIAN TUBES)
The oviducts are a pair (left and right) of
muscular tubes (about 12ern long) that serve as --
passageways for the ovum or the conceptus on
its way to the uterus, and for the sperms on their
way to fertilizing the ovum.
Grossly, an oviduct has four segments:
1) infundibulum-the funnel-shaped area
that is related to the ovary and that opens
into the peritoneal cavity and whose margins
are provided with numerous processes
called fimbriae; 2) ampulla-the expanded
intermediate portion that comprises 2/3 of the
length of the tube; 3) isthmus-the narrow and
slender part that connects the Fallopian tube to
4;\ 1}a.u Interstittalis-c- the 1]act
tb,.l!.1J.tl!.~1J.~)a.tAd
of the tube within the uterine wall.
Histologic Layers of the Oviducts
Histologically, an oviduct has three layers,
from within outwards these are mucosa,
muscularis and serosa.

The oviductal mucosa, which consists of
an epithelium and lamina propria, forms
numerous folds in the ampullary area. But as
one goes proximally, the mucosal folds become
much shorter and less complex. In. the pars
interstitialis, they merely form ridges.
The epithelium of the oviduct is generally
simple columnar. The cells are tall in the
ampulla but diminish in height towards the
uterus. Two types of cells are distinguishable
in the epithelium. One type is ciliated. The
ciliated cells are numerous in the infundibulum
and in the ampulla and their cilia beat toward
the uterus. They play an important role in
transporting the ovum or conceptus to the
uterus. The other cell type is called peg cell.
It is nonciliated but secretory. The secretion of
Fig. XVII-13. Oviduct (A). The diagram shows
the histologic layers of the organ.
Fig. XVII-1.4. Fallopian Tube (B). This section
of the ampullary region of the organ shows a
mucosa that forms numerous intricate folds.
External to the mucosa is muscularis layer (m)
and a serosa (5). H&E x40.
Fig. XVII-1S. Fallopian Tube (C). The mucosa
of the oviduct consists of a ciliated simple
columnar epithelium (e)and an underlying lamina
propria (lp), The muscularis layer is made up two
layers of smooth muscle cells. In the inner layer
(im), the muscle cells are circularly- or spirally-
arranged, while in the outer layer (om),they are
longitudinally-arranged.The outermost histologic
layer of the organ is serosa (5). H&E x100.
peg cells provides the conceptus with nutritive
material as it traverses the length of the oviduct
on its way to the uterus.
The lamina propria of the oviduct is very
cellular connective tissue that is richly supplied
with blood and lymphatic vessels. It is devoid of
true glands.
The muscularis of the oviduct consists of
an outer layer of longitudinally arranged and
an inner layer of circularly or spirally arranged
smooth muscle cells that gather in bundles. In
between the muscle bundles is an abundant
amount ofloose connective tissue.
The outermost histologic layer of the
oviduct except for that in the pars interstitialis,
which is within the uterus, is serosa. It contains
a plexus of nerves from where fibers pass inward
FEMALE REPRODUCTIVESYSTEM +B
to supply the muscle and mucosal layers. Like
the lamina propria, the serosa is richly supplied
with blood and lymphatic vessels.
Effects of Ovarian Hormones on
the Oviducts
During the first half of the menstrual cycle,
the ciliated cells of the oviductal epithelium
increase in number and height in response
to the estrogen that comes from the ovarian
follicles.
At about mid-menstrual cycle, just about
the time of ovulation, the ciliated cells are at
their tallest and their ciliary beat at their fastest
while the peg cellsbecome secretory.
Later in the cycle, progesterone favors the
loss of cilia and a decrease in the secretory
activity of the epithelial cells. During the last
few days of the cycle, very few cells of the
oviductal epithelium are still ciliated and many
of the secretory cells are already atrophic.
UTERUS
The uterus is a pear-shaped, dorsoventrally
flattened, hollow, pelvic organ that receives
the conceptus a few days after fertilization
and nourishes and nurtures it throughout its
development. In the non-pregnant adult female,
it is about 6.5 cm long, 3.5 em wide, and 2.5 ern
thick.
The uterus has two regions: an expanded
upper region comprising much of the organ
referred to as body or corpus uteri, whose
cavityis called uterine cavity, and a cylindrical
inferior region referred to as cervix, whose
cavity is called cervical canal (endocervical
canal). The two regions are demarcated
from each other by a constricted area, the
isthmus. The part of the corpus uteri above the
attachment ofthe oviducts is called fundus. The
uterine cavity is continuous with the cavity of
the vagina (vaginal canal) inferiorly through
the cervical canal.
L;TEB/,f\: (!, CiO\iZ/LES' TEXTBOOK OF HISTOLOGY
Corpus Uteri
Three histologic layers are distinguishable
in the corpus uteri. The innermost layer is a
mucosa called endometrium. The middlelayer,
the myometrium, is the thickest of the three
layers and it consists mainly of smooth muscle
cells. The outermost layer, which is referred to
as perimetrium, is serosa/adventitia.
Endometrium
The endometrium is where the conceptus
implants and develops initially into an
embryo and later into a fetus. It consists of
an epithelium and an underlying lamina
propria. The endometrial epithelium is simple
columnar and some of the cells are ciliated.
From the epithelium, simple tubular glands
(endometrial glands) invaginate into the entire
thickness of the lamina propria where they are
embedded in a connective tissue stroma that
has an abundance of reticular but a paucity of
elastic fibers. The cells in the lamina propria
ace mainly fibroblasts but lymphocytes
and other types of leukocytes are also -.
present.
The endometrium varies in thickness from
0.5 to 6.0 mm during the course ofthe menstrual
cycle. Its superficial portion, which comprises
2/3 of the endometrium when it is at its thickest,
is shed during menstruation and is aptly referred
to asfunctional layer while its deeper 1/3, which
is responsible for regenerating the functional
layer after menstruation is referred to as basal
layer.
The blood supply of the endometrium
comes from branches of the uterine arteries
called arcuate arteries. T~e arcuate arteries,
after penetrating the myometrium, give off
branches that supply the basal layer of the
endometrium but the main vessels continue
inward to supply the functional layer. In the
functional layer, the branches of the arcuate
arteries are very tortuous and are called coiled
or helicine arteries.
Endometrial Cycle
The endometrial cycle refers to the
recurring morphological and physiological
changes that the endometrium undergoes
during the menstrual cycle in response to the
ovarian hormones. These changes are designed
to ensure that the endometrium is prepared for
implantation of the conceptus in the event of a
successful fertilization.
The endometrial cycle is divided into
three stages: menstrual phase, proliferative
or follicular phase, secretory or luteal phase,
and menstrual phase.
The proliferative phase is governed by
estrogen that is secreted by the ovarian follicles.
In relation to the ovarian cycle, it coincides
with the growth of the ovarian follicle. The
secretory phase starts after ovulation and is
governed by progesterone that is produced by
the corpus luteum in' the ovary. The menstrual
phase, meanwhile, results from the withdrawal
of the ovarian hormones during the latter part of
the menstrual cycle.
" over the denuded endometrium. When re-
Proliferative Phase (Follicular Phase)
The proliferative phase of the endometrial
cycle starts at the end of menstruation (i.e.,about
3 to 5 days into the menstrual cycle)-when
the endometrium consists simply of the basal
layer and is merely 0.5 mm to 1.0 mm thick-
and ends at ovulation. During this phase, the
functional layer of the endometrium which
Fig. XVII-16. Early Proliferative Endometrium.
In this section, the endometrium has been partly
restored. The epithelium (e) has been completely
re-established and some glands (g) have formed.
Deep in the endometrium is the myometrium
(my). Uterus, H&E x100.
has been shed off in the preceding few days (i.e.,
during the menstrual phase) is restored.
Shortly after the menstrual phase has
begun, the cells in the remaining portions of
the glands in the basal layer of the endometrium
start dividing to form a surface epithelium
epithelialization of the endometrium has been
completed, menstruation ends, and circulation
in the superficial areas of the endometrium is
re-established and the proliferative phase starts.
During the proliferative phase, the
endometrial mucosa undergoes several fold
increase in thickness because of continuous
cellular proliferation and activity i 1 the glands
and stroma. The glands increase markedly in
number and in length; the ground substance
becomes abundant; and the blood vessels
elongate and become tortuous. Towards the
end of this phase, just before ovulation, the
glandular cells start to accumulate glycogen.
In routine LM preparations of the
proliferative endometrium, many mitotic
figures are seen in both glandular and stromal
Fig. XVII-17. Proliferative Endometrium. This
section shows an endometrium that is almost
completely restored. The glands (g) are already
rather long and the lamina propria (lp) under the
epithelium (e) is copious. Uterus, H&E x40.
FEMALE REPRODUCTIVESYSTEM ..
cells and the endometrial glands are noted to be
relatively straight and smooth contoured and
with narrow lumens.
Secretory Phase (Luteal Phase)
The secretory phase of the endometrial
cycle starts immediately after ovulation and
lasts until the start of the menstrual phase of
the next menstrual cycle.
Fig. XVII-18. Secretory Endometrium. Note
the abundant lamina propria (lp) that underlies
the epithelium (e). The glands (g) look like
"corkscrews" and some show secretory-material
in their lumen. Uterus, H&E x100.
During the secretory phase, the endometrial
glands become tortuous and secretory, while
the coiled arteries elongate further and become
more convoluted. The endometrium is at its
thickest during this phase, but its increase in
thickness is not due to mitoses because hardly
any is occurring, but rather to the hypertrophy
of the glandular cells caused by accumulation
of glandular secretion, edema, and increased
vascularity of the connective tissue stroma.
Thus, in routine LM preparations of the
secretory endometrium, the glands are noted to
be large and highly coiled (corkscrew-shaped)
and their lumens filled with secretions.
The secretion of the endometrial glands
consists of a carbohydrate-rich fluid that
serves as nourishment for the embryo before it
implants.
ESTE8r\N& GONZALES'TEXTBOOK OF HISTOLOGY
Menstrual Phase
At about day 25 or 26 of the menstrual
cycle, in the absence of fertilization and
implantation, the blood levels of the ovarian
hormones (l.e., estrogen and progesterone)
drop. This drop results in marked vascular
changes in the endometrium. The coiled
arteries constrict intermittently for variable
periods of time, compromising the blood
supply of the functional layer. Consequently,
the glands stop their secretory activity and
degenerate. After about a couple of days of
intermittent constriction, the coiled arteries
close down completely for several hours. This
causes necrosis of the whole functional layer.
When the arteries reopen, the damaged vessels
in the functional layer burst and blood pours
into the necrotic area, which then exfoliates and
flows out of the uterus with the extravasated
blood, heralding the start of the menstrual
phase of the next menstrual cycle.
The average blood loss per menstrual flow
is only about 35 mL to SOmL. However, the
volume of menstrual flow is much more than
SOmL because it includes not only arterial and
venous blood, but also epithelial and stromal
cells, and the secretions of the uterine and
cervical glands.
Myometrium
The myometrium, the thickest layer of the
corpus uteri, is made up of bundles oflarge and
long (40 ~m to 90 um) smooth muscle fibers.
The muscle bundles are separated by connective
tissue and form four ill-defined layers.
The myometrial muscle fibers increase in
size in the presence of estrogen and are smallest
immediately after menstruation.
In pregnancy, when the uterus increases in
size dramatically, new muscle cells are formed
and the existing ones enlarge and lengthen to as
long as 500 ~m.At the same time, the connective
tissue between the muscle fibers and bundles
also increases in amount.
 After parturition (i.e., delivery), most of
the muscle cells go back to their original size
while some degenerate.
Perimetrium
The perimetrium is a typical serosa that
consists of a thin loose connective tissue layer
that is overlaid by mesothelium over the fundus
and much of the posterior aspect of the uterus
where the organ is covered by peritoneum, but
elsewhere, there is no mesothelium and the layer
is simply an adventitia.
Cervix
The cervix is the cylindrical inferior
portion of the uterus whose proximal end
projects into the vagina. The part of the cervix
that protrudes into the vagina is referred to
as portio vaginalts (exocervix; ectocervix),
while the rest of the cervix is called endocervix.
Its cavity, the cervical canal (endocervical
canal), communicates superiorly with the
uterine cavity via the internal os and inferiorly
with the vaginal canal through the external os,
Although part of the uterus, the cervix markedly
differs from the corpus uteri in structure and
function.
Fig. XVII-19. Endocervix. Note that the mucosa
forms folds in this section of the cervix. The
epithelium (e) is simple columnar. It invaginates
into the lamina propria (Ip) to form branched
tubular cervical glands (cg). Cervix, H&E x100.
Histologically, the innermost layer 'of the
cervix is a mucosa that forms folds called plicae
palmatae in the cervical canal. It consists of an
epithelium and a lamina propria. The epithelium
is simple columnar in the cervical canal (like in
the corpus uteri), but nonkeratinized stratified
squamous in the portio vaginalis. The transition
between the two types of epithelia occurs
abruptly at or near the external os.
Fig. XVII-20. Transitional Area between
Endocervix and Exocervix. Note that at the
region indicated by the unlabeled arrow, the
simple columnar epithelium that characterize
the endocervix abruptly changes into the
nonkeratinized stratified squamous epithelium
of the portio vaginalis of the cervix (exocervix).
Also take note of the cervical glands (cg) in the
lamina propria of the endocervix. H&E x100.
The lamina propria, especially in the
area of the external os, is heavily infiltrated
with lymphoid elements. It contains mucus-
secreting tubular glands (cervical glands) that
branch extensively. The ducts of the glands
often get clogged and the glands become cystic
(Nabothian cysts).
External to the mucosa, the substance of the
cervix, unlike that in the corpus uteri, contains
very few smooth muscle fibers. It consists
mainly of collagenous connective tissue that is
well-supplied with elastic fibers.
In the cervix, the epithelium does not
exfoliate. The cyclical changes that occur in
 response to the ovarian hormones are more
pronounced in the glands and connective tissue
stroma. The secretion of the cervical glands is
copious, thin, and watery during mid-cycle (i.e.,
about the time of ovulation), but viscous during
the rest of the cycle. The character and quantity
of the secretion of the glands during the mid-
menstrual cycle is favorable to the migration of
the sperm cells.
During parturition, the cervix forms part
of the birth canal. At other times, it possibly
helps prevent the entry of microorganisms
into the uterus. The secretions of its glands
help lubricate the vaginal canal and facilitate
the passage of the spermatozoa into the upper
genital tract.
VAGINA
- due to transudation from the capillaries in the
The vagina is a fibromuscular tube that
extends from the vestibule of the external
genitalia to the cervix.Normally, it is a collapsed
tube and its anterior and posterior walls are in
contact with each other.
The vaginal wall has three histologic layers:
, mucosa, muscularis, and adventitia.
Fig. XVII-21. Vagina. The photomicrograph
shows the vaginal mucosa made up of a
non keratinized stratified squamous epithelium
(e) and lamina propria (lp), and the muscularis
layer (m). The lamina propria of the vagina is
highly vascular as attested to by the numerous
arteries (a) and veins (v). H&E x100.
ESTEBAN & CCI\lD\LES' TEXTBOOK OF HISTOLOGY
The mucosa, which is thrown into
transverse folds called rugae, consists of a
surface epithelium and lamina propria. The
epithelium is continuous with and similar to the
epithelium of the portio vaginalis of the cervix,
,i.e., nonkeratinized stratified squamou . The
lamina propria is made up of highly vasculan
dense connective tissue that contains many
elastic fibers and MALT that include occasional
lymph nodules.
The vagina has no glands. The epithelial
surface is kept moist by secretions coming from
the cervical and endometrial glands.
Incidentally, the increase in the amount
of fluid that occurs in the vaginal canal in
response to erotic stimulation cannot be totall
accounted for by the secretion of the cervical
and endometrial glands. Evidently, some of it is!
lamina propria.
The vagina has no submucosa. The lamina;
propria blends with the muscularis, which is
composed of smooth muscle fibers that are
arranged into two poorly-defined layer&-an
inner layer where the muscle fibers are circularly
arranged and an outer layer where the muscle
fibers are longitudinally arranged.
The adventitia of the vagina is thin. It is
made up of dense connective tissue that blends
with the loose connective tissue that connects
the vagina to the surrounding structures. It also
contains nerve bundles and an extensive venous
plexus.
The vagina undergoes minor changes in
response to the ovarian hormones, but notably,
the desquamation of the epithelial cells is
greater during the second half of the menstrual
cycle and during menstruation than during the
first half of the menstrual cycle.
EXTER Al GE ~TALlA(VULVA)
The external genitalia are the structures of
the female reproductive system that are external
to the vagina. They include the clitoris, labia
majora, labia minora and the minor and major
vestibular glands whose ducts open into the
vestibule of the external genitalia.
The clitoris is the homologue of the penis.
It consists of two cavernous bodies (corpora
cavernosa) that are made up of erectile tissue
.nipple
and that both end distally in a small rounded
structure, the glans clitoridis. The clitoris is
covered by stratified squamous epithelium and
has numerous specialized nerve endings. areola

The labia minora are membranous folds

that form the lateral walls of the vestibule. They
are made up of a stratified squamous epithelium
and an underlying highly vascular connective
tissue that contains numerous sebaceous
glands, but has no hair follicles.
The labia majora cover the labia minora.
They are the homologues of the scrotum and
are basically made up of highly pigmented skin
except in their inner surfaces where they are
smooth and hairless.
Several (minor) vestibular glands are
located around the urethral opening and near
the clitoris. They are mucus-secreting and
resemble the urethral glands (of Littre) of the
male.
The major vestibular glands (Bartholin's
glands) are a pair of larger glands in the
lateral walls of the vestibule. They are about
1.0 ern each in diameter and they open on the
inner surface of the labia minora. They are
branched tubuloalveolar and mucus-secreting,
and resemble the bulbourethral glands (of
Cowper) of the male.
IMAMMARY GLAND (BREAST)
In humans, a pair of mammary glands is
normally present in both sexes,but they undergo
marked development only in the female and
only starting at puberty.
The mammary glands of females attain
their highest degree of development during and
immediately after parturition. The mammary
glands are modified apocrine sweat glands
that, in mammals, have evolved to provide
Fig. XVIl-22. Mammary Gland
nourishment for their offspring during the
initial stages of growth and development.
lobes of the Mammary Gland
In the sexually mature female, the
mammary gland is made up of IS to 20 lobes
that are separated from each other by interlobar
connective tissue. Each lobe is drained by
a single lobar duct (lactiferous duct) that
opens into the nipple independent of the other
lactiferous ducts. Thus, the mammary gland can
be regarded as consisting of IS to 20 separate
compound tubuloalveolar glands.
The interlobar connective tissue sends
connective tissue septa into the lobes to divide
each lobe into lobules. The lobules consist of
secretory alveoli and tubules, and their ducts
that are supported by a connective tissue stroma.
The amount of connective tissue and the
number and sizes of the alveoli and ducts in the
lobules depend on the state of activity of the
organ.
The secretory alveoli and tubules are
drained by intralobular ducts that unite to
form interlobular ducts which in turn merge to
form the lactiferous duct. Near its termination,

FEMALE REPRODUCTIVESYSTEM +a
the lactiferous duct has a dilated portion called
lactiferous sinus. The lactiferous duct and
sinus are lined by stratified cuboidal epithelium.
Areola and Nipple
The nipple is a small skin protrusion on
the mammary gland that contains the orifices
of the lactiferous duct. The skin of the nipple
is highly pigmented and richly supplied with
sensory nerves. The dermis of the nipple has
many smooth muscle fibers.
The nipple is surrounded by a highly
pigmented area of skin, the areola, whose
dermis contains some hair follicles, sweat
and sebaceous glands, and the glands of
Montgomery-large, branched glands of
the apocrine type that, structurally, are a
cross between a sweat gland and a mammary
gland.
Fig. XVII-23. Inactive Mammary Gland. The
glandular tissue (g) consists mostly of ductal
elements. There is also an abundance of adipose
tissue (a). H&E x100.
Inactive Mammary Gland
The bulk of the inactive mammary gland
is connective tissue, much of which is adipose
tissue. In fact, the amount of adipose tissue
in an inactive mammary gland determines its
size. Alveoli, if present, are in the form of small
buds. The ductal system is, however, extensive.
The lining epithelium is simple cuboidal in the
small ducts but progressively grows taller until
it becomes stratified in the main ducts.
[STES;\N & GONZ!\LES' TEXTBOOK OF HISTOLOGY
Mammary Gland in Pregnancy
During pregnancy, the cells of the
intralobular ducts proliferate and give rise to
alveoli. At the same time, the interlobular and
intralobular connective tissue decreases in
amount to give way to the expansion of the
lobules. In the latter part of pregnancy, the
alveolistart to elaborate some secretory material
called colostrum; and the nipple and the areola
become highly pigmented.
Lactating Mammary Gland
In the lactating breast, the alveolar cells
undergo a cyclicalprocess of secretion and rest.
Typically, many of the alveoli are dilated and
distended by milk so that their epithelium is
flattened while many other alveoli are resting
and have smaller lumens and taller epithelial
cells.
The initial secretory product of the
lactating breast is referred to as colostrum.
Aside from nutrients, colostrum contains
immunoglobulins that confer passive
immunity on the newborn. A few days after
parturition, the secretion of colostrum by the
alveoli of the mammary gland stops, giving
way to the secretion of true milk.
In the mammary glands, the versatile
cells of the alveoli produce all the constituents
of milk. The synthesis of the protein
components is merocrine in nature. The
proteins are synthesized in the granular
endoplasmic reticulum and packaged into
small vacuoles within the Golgi complex. The
packaged secretory materials then move to
the apical cytoplasm where they are released
by exocytosis. The production of the fat
components, on the other hand, is apocrine in
nature. The fats start as small droplets within
the cytoplasm. These droplets later coalesce to
form larger fat globules that move to the apex
of the cell. When the globules are released, the
apical cell membrane and some cytoplasm go
with them (apocrine secretion).

Fig, XVII-24-, Lactating Mammary Gland,
Connective tissue septa (s) divide the gland
into lobules. The lobules are filled with dilated
and distended alveoli (a), many of which contain
secretory material in their lumen. In the septa,
some interlobular ducts (ild) can be seen. H&E
x100.
In the alveoli and small ducts of the
mammary gland, myoepithelial cells are
present between the lining epithelial cells
and the basal lamina. Their contraction helps.
in expressing milk from the alveoli and small
ducts.
Mammary Gland after Weaning
and Menopause
After the baby has been weaned, the
mammary gland regresses and reverts to its
inactive state. However, the gland usually does
not return completely to the pre-pregnant state
because many of the alveolipersist.
After menopause, the mammary gland
involutes. The alveoli disappear completely.
Likewise, the duct system progressively
dissipates such that only very few ducts persist
into old age.The connective tissue and adipose
tissue likewise progressively diminish in
amount.
Implantation and Placentation
If the ovum released into the abdominal
cavity during ovulation is fertilized, the
resulting conceptus (zygote) immediately
travels to the uterus and undergoes a series of
mitotic divisions (cleavage).
By the time it reaches the uterine cavity
about three to four days after fertilization, the
conceptus already consists of a lump of cells
(blastomeres) that looks like a mulberry and is
called a morula.
In the uterine cavity, the conceptus floats
freely, subsisting on the secretions of the
uterine glands, for another two to three days.
During this time, it transforms from a morula
into a fluid-filled sphere called blastocyst,
which consists of a) an enclosing wall known
as trophoblast that is formed by a Singlelayer
of cells; and, b) an aggregation of cells called
inner cell mass on one pole (embryonic
pole) of the blastocyst. The inner cell mass
gives rise to the embryo proper while the
trophoblast forms the covering membranes
of the fetus and participates in the formation
of the placenta.
IMPLANTATION
About six to seven days after fertilization,
implantation, which refers to the embedding
of the blastocyst into the endometrium,
starts. At this time, the endometrium is in the
secretory or luteal phase and is at its thickest.
It is very edematous and the glands are actively
secreting. Implantation in humans is referred
to as interstitial implantation, which involves
complete embedding of the embryo within the
endometrium.
At the start ofimplantation, the trophoblast
at the embryonic pole of the blastocyst attaches
to the luminal surface of the endometrial
epithelium of the anterior or posterior wall
(usually towards the fundus) of the uterus.
Then, the cells of the trophoblast that get in
contact with the endometrium proliferate
rapidly and soon two distinct layers can be
distinguished in the trophoblast: an outer layer,
the syncytiotrophoblast, a multinucleic layer
without cellboundaries; and an inner layer,the
FEMALE REPRODUCTIVESYSTEM ..
cytotrophoblast (Langerhans layer), which is
made up of cells that are distinctly separated by
cell membranes.
The syncytiotrophoblast secretes enzymes
that erode the endometrium and enable the
conceptus to embed slowly into the endometrial
stroma.
By the ninth to 11th day after fertilization,
the conceptus, which at this time has two
germ layers in the embryo proper already,
has completely embedded in the endometrial
stroma. The defect in the endometrial surface
that resulted from the entry of the conceptus
is temporarily sealed by a closing coagulum
made up of fibrin and cellular debris. Later, the
epithelium is restored.
PLACENTATION
After implantation, 'an elaborate system
of delivering nutrients and oxygen to the
embryo is required to support its rapid growth
and development. This is the reason why the
placenta is formed.
The placenta is offetal and maternal origin,
but more of the former. When fully developed,
it is a flat, discoid organ that measures about 15
cm in diameter and 3 em in thickness.
During pregnancy, the blood of the fetus
and that of the mother do not mix and do not
come in actual contact with each other. Instead,
substances in the blood of the mother diffuse
into the blood of the fetus, and vice-versa; via
the placenta.
Fetal blood -is brought to and from the
placenta by the blood vessels of the umbilical
cord. At birth, the placenta is expelled from
the uterus several minutes after the fetus has
been delivered. After delivery of the fetus, the
umbilical cord is normally ligated and cut by the
birth attendant (often, a physician) before the
placenta is delivered.
The placenta is also an endocrine organ.
It produces several hormones that are vital to a
successful pregnancy and delivery.
ESTEBi1J4 & GONZ!\LES' TEXTBOOK OF HISTOLOGY
Development of the Placenta
During the second week after fertilization,
cavitiesappear within the syncytiotrophoblast.
These cavities are called lacunae. They
soon get filled with blood from maternal
capillaries that have been eroded by the
syncytiotrophoblast. The lacunae progressively
increase in size until only cords of trophoblast
(i.e., a core of cytotrophoblast covered by
syncytiotrophoblast) can be seen dipping
into them. These trophoblastic cords cover
the whole conceptus and are termed primary
trophoblastic villi.
At about the 15th day after fertilization,
loose connective tissue from the
extraembryonic mesoderm forms a lining
layer on the inner aspect of the trophoblast.
This extraembryonic mesoderm and the
trophoblast are collectively referred to as
chorion. Mesodermal tissue soon penetrates
the cores of the primary trophoblastic villi,
and at this point, the villi are referred to as
secondary villi.
Blood vessels later differentiate from the
mesodermal tissue at the cores of the secondary
villi. Shortly thereafter, the blood vessels fuse
-I
Fig. XVII-2S. Umbilical Cord. The umbilical
cord consists of mucous connective tissue called
Wharton's jelly (wj) where three blood vessels-
two umbilical arteries (a) and an umbilical vein
(v}-and a small tubular structure called allantois
(al), which is vestigial in humans, are embedded.
H&E x5.

Fig. XVII-26. Placenta. Photomicrograph
shows cross section of placental villi (Vi) that
are enveloped by trophoblast (T). The villi
contain numerous capillaries (C). In life, they are
immersed in maternal blood that fills the lacunae
(L). H&E x400.
to form a continuous vascular channel from the
placenta to the fetal heart, and vice-versa. By
the end of the third week of development, fetal
blood already circulates within the capillaries
of the villi.
Once a villus has acquired a blood vessel,
it becomes known as a tertiary or definitive' ,
villus. Initially, definitive villi completely
surround the embryo.
Decidua and Chorion Frondosum that is lined by typical endothelium. It is also
During implantation, as previously
mentioned, the conceptus embeds entirely
within the endometrium, which during
pregnancy is referred to as decidua.
When implantation has been completed, the
portion of the endometrium that underlies the
implantation site is called the decidua basalis.
The endometrial tissue between the implanted
product of conception and the uterine cavity is
called the decidua capsularis. The rest of the
endometrium is called the decidua vera.
As the fetus grows within the endometrium,
it bulges into, and later obliterates, the uterine placenta, produces several hormones that
cavity. In the process, the decidua capsularis
meets and merges with the decidua vera of the The more important of these hormones are
opposite wall of the uterus.
When definitive villi appear, they 'are
initially equally distributed all around the
embryo. However, at about the eighth week
of development, the definitive villi associated
with the decidua capsularis degenerate leaving
a bald area called chorion laeve. Meanwhile,
the definitive villi associated with the decidua
basalis enlarge, rapidly increase in number, and
branch extensively. These definitive villi are
collectively referred to as chorion frondosum.
The chorion frondosum comprises the
fetal contribution while the decidua basalis
represents the maternal contribution to the
definitive placenta.
Histology of the Placenta
The fully formed placenta is partitioned
into 15 to 20 parts (cotyledons) by incomplete
septa that project from the decidua basalis
into the intervillus spaces. Each cotyledon is
supplied with a large definitive villus that has
several generations of branches.
Terminally, placental villi have identical
histologic characteristics. They have a core
of loose connective tissue that contains some
smooth muscle cells and phagocytes. The
connective tissue core is supplied with a capillary
enveloped by the two layers of the trophoblast,
i.e., cytotrophoblast and syncytiotrophoblast.
The cytotrophoblast degenerates as the
pregnancy nears term. At delivery, it has mostly
disappeared and only a few cytotrophoblast
cells persist internal to the syncytiotrophoblast.
Placental Hormones
The syncytiotrophoblast, which, as
previously discussed, appears in the developing
conceptus when it starts to implant and later
becomes part of the developing and definitive
are essential for a successful pregnancy.
human chorionic gonadotropin (heG),
FEMALE REPRODUCTIVESYSTEM +M
progesterone, estrogen, and, human
chorionic somatomammotropin (hCS;
chorionic growth hormone-prolactin; CGP;
human placental lactogen). In some mammals,
but probably not in humans, relaxin is also
secreted by the placenta.
Human chorionic gonadotropin (hCG)
is a glycoprotein hormone that is detectable
in maternal blood as early as six days after
fertilization. Itis excreted via the kidneys and its
presence in urine very early in pregnancy forms
the basis of many pregnancy tests.
Human chorionic gonadotropin (hCG)
is similar in chemical structure and function
to LH. It enlarges and transforms the corpus
luteum in the ovary to a corpus luteum of
pregnancy.
ESTEBAN& GONz/\LES' TEXTBOOK OF HISTOLOGY
By the sixth week of gestation, the placenta'
produces enough estrogen and progesterone
eighth week of pregnancy, the corpus luteum
of pregnancy starts to decline functionally, but
it persists for several months (sometimes up to
term) although no longer needed.
The maternal requirement for estrogen and
progesterone during pregnancy is so huge that
the blood levels of these hormones at term are
several-fold higher than during the luteal phase
of the menstrual cycle.
Human chorionic somatomammotropin is
a protein hormone that has a similar structure
to human growth hormone and probably
functions as a "maternal growth hormone of
pregnancy."
Endocrine
System

he endocrine system and the nervous messengers. Hormones are secreted into

T system comprise the two major capillariesand carried by blood to target organ/s
integration and control systems of the or tissue/s that contain/s the cells (target cells)
body. The two systems are able to receive and that possess appropriate receptors for them.
respond to stimuli, but whereas the nervous The endocrine cells in the body occur: 1)
system responds rapidly and precisely to embedded singly or in clusters in the various
stimuli, the endocrine system responds more organs such as the JG cells in the kidneys,
slowly and more diffusely although the effects enteroendocrine cells in the gastrointestinal
of its responses are usually longer lasting.' . tract, and interstitial cells (of Leydig) in the
Furthermore, many of the responses of the testes; 2) as distinct organs called endocrine
nervous system are consciously generated, but glands, i.e., pituitary, thyroid, parathyroid,
the responses of the endocrine system are all adrenal, and pineal; and; 3) as well-defined
done unconsciously. component structures of certain organs, i.e.,the
As a rule, the nervous and endocrine endocrine portion ofthe hypothalamus and the
systemsworkin parallelwith, but independently islets of Langer hans of the pancreas.
of, each other. However, some anatomic and
functional overlaps exist between the two
systems. Someparts of the nervous system, such
as the hypothalamus, are also endocrine organs.
Likewise, there are certain endocrine responses
that are generated by nervous impulses and vice-
versa.

The endocrine system is composed of

all the endocrine cells in the body, i.e., cells
that produce substances (steroids, peptides or
amines) called hormones that act as chemical Fig. XVIII-1. Hypothalamus and Pituitary Gland
 The hormone-producing cells that are
embedded in the various organs have been
discussed in connection with the organs where
they are located. Hence, the discussion will
be limited to the endocrine portions of the
hypothalamus and pancreas, and the endocrine
glands-glands that do not poss~ss any
excretory duct, but which are richly supplied
with blood capillariesinto whichtheir secretions
diffuse.
YPOTHA A U
The hypothalamus is part of the brain. It
is a component of the diencephalon,' which
together with the telencephalon comprises
the forebrain or prosencephalon. The
hypothalamus is located at the base ofthe brain,
behind the optic chiasm,' and it forms the floor
and part of the wall of the third ventricle. It
consists of 11 major nuclei, nuclear areas and
tracts.'
The hypothalamus controls numerous
bodilyfunctions includingmanyvegetativeones
such as thirst, hunger and satiety, temperature,
sexual behavior, and circadian rhythms. In
addition, it produces two groups of hormones:
posterior pituitary and hypophysiotropic. '
Posterior Pituitary Hormones-
The posterior pituitary hormones are
oxytocin and antidiuretic hormone (ADHj
vasopressin), They are synthesized by neuron
(magnocellular secretory neurons) that
are in the supraoptic and araventricular
nuclei of the ypothalamus, but stored in the
posterior lobe of the pituitary glan~, thus,
their collective name. The supraoptic nucleus
is located above and lateral to the optic chiasm
whil1 e th e paraven trincu 1ar nuc 1eusf· IS In
. th e
the hypothalamu but they are mostly in th
thi d tri 1 1\ t . • arcuate, paraventricular and periventricular
1at era 1 wa 110f th e If ven nc . vxy, OG-In
. pro d uce d b y one ce 11 typ wh'l1 e AT'\H
IS '.lJ nuclei'
is produced by another cell ty e, but bot The function of the hypophysiotropic
cell types are present in the supraoptic an hormones is to re ulate the activity of th
paraventricular nuclei. secretory cells of the pjtJJjla-qLgla:ndP
The main target organs of oxytoci in
women are the uterus and the breasts. In the
uterus, it stimulates contraction of the smoot
muscles in the myometrium d thus, aids in
parturition i.e., delivery of the fetus). In the
mammary glands, it promotes ejection of mil
during lactatio by stimulat,ing contraction 0
the myoepithelialcellsthat surround the alveoli.
Oxytocin's function in non-pregnant women
has not been definitely established yet but it
may playa role in various social behaviors and
evenwound healing. In males, current evidence
suggests that oxytocin facilitates sperm
transport within the male reproductive system.
The target organs of ADH, on the other
hand, are the kidneysflt increasespermeability
of the distal and collecting tubule of the
kidneys, leading to the formation of more
concentrated urine.
I;IYPDphys.iotropicHormones
The hypophysiotropic hormones
(ileurohormones), hose names describe their
main functions, include: 1) codk9trophin-
releasing hormone (CRH),'2) thyrotropin!
releasing hormone (TRH), 3) growth'
hormone-releasing hormone (GRH), 4)
gonadotropin-releasing hormone (GnRHj
luteinizing hormone-releasing hormone,
LHRH), 5) growth hormone-inhibiting
hormone (GHIHj somatostatin, SSI), and 6)
prolactin-inhibiting hormone (PIH).
The hypophysiotropic hormones
are synthesized by secretory neurons
(parvocellular secretory neuro!s) that are
smaller than the neurons that produce the
posterior pituitary hormone .The parvocellular
secretory neurons are widelydistributed within

ESTEBI\[\I& GCNb'\LES' TEXTBOOK OF HISTOLOGY

PITUITARY GL ND
pars tubatal1$
(HYPOPHYSIS CEREB I)
pituitary stalk
The pituitary gland is a small but very
important endocrine gland. It is an ovoid body
that weighs merely SOOmg and is only about 12
mm in transverse and 8 mm in AP diameters.
It is attached to the inferior surface of the
hypothalamus and is lodged in a concavity
(hypophyseal fossa) in the sella turcica of the
sphenoid bone.
The pituitary gland consists of two
distinct parts, neurohypophysis and
adenohypophysis. These two parts
differ developmentally, structurally, and
functionally.

magnocellularo;:,_ .. 'I.C''''' 1

neurons -

Fig. XVIlI-3. Pituitary Gland. The scanning

photomicrograph shows the posterior lobe
and pituitary stalk of the neurohypophysis; and
the anterior lobe, intermediate lobe, and pars
" tuberalis of the adenohypophysis. Pituitary
Gland, H&E x40.

the other hand, arises from oral ectoderm, it

is derived from Rathke's pouch, an upgrowth
of the oral mucosa. During embryonic
development, the two ectodermal outgrowths
meet and fuse to form the pituitary gland. The
connection of Rathke's pouch to the oral cavity
is subsequently severed, but the connection of
the neurohypophysis to the base of the brain,
specifically to the hypothalamus, persists.

Fig. XVIII-2. Pituitary Gland
Development of the Pituitary
Gland
The neurohypophysis arises from neural
ectoderm, it develops as a downgrowth of
the diencephalon. The adenohypophysis, on
Neurohypophysis
The neurohypophysis has three regions:
'1) median eminence, 2) pituitary stalk
(infundibulum, infundibular s t em:
infundibular stalk, hypophyseal stalk) j and
3)posterior lobe (pars nervosa, infundibular
process).
The median eminence is the proximal
portion of the neurohypophysis that is attached

ENDOCRINE SYSTEM Wi
---- ---- ------------------------------------------------------~
to the hypothalamus. The slender downward The axons of the magnocellular secretory
continuation of the median eminence is the neurons have dilatations, not just at their ends
pituitary stalk while the expanded inferior but along their entire length. The dilatations are
continuation of the pituitary stalk is the posterior the storage sites for the secretory granules that
lobe. Incidentally, some authors consider the are synthesized in the cell bodies of the neurons.
median eminence as the most inferior part of They are associated with capillaries into
the hypothalamus, instead of being the proximal which their secretions ate released, as needed.
portion of the neurohypophysis. Aggregations of secretory granules in the
The easiest way to understand the histology axonal dilatations are often seen in routine LM
of the neurohypophysis is to simply consider it as preparations of the posterior lobe and pituitary
the downward extension of the hypothalamus. stalk as 'deeply-staining, basophilic structures
Why and how is this so? called Herring bodies. The posterior lobe is
much bigger than the pituitary stalk simply
The cell bodies of the secretory neurons because most of the axonal dilatations are in
in the hypothalamus (i.e., parvo cellular the former.
and magnocellular secretory neurons) are
located in the nuclei and nuclear areas, but In the neurohypophysis, the axons of the
their unmyelinated axons travel downwards, secretory neurons are surrounded by numerous
leave the hypothalamus and terminate in the non-secretory supporting cells called pituicytes
neurohypophysis. These axons comprise the that are morphologically similar to astrocytes.
bulk of the neurohypophysis. Aside from the Pituicytes are stellate cells whose slender
axons, the only other structures that are present cytoplasmic processes are interconnected with
in the neurohypophysis are capillaries and
those of other pituicytes by gap junctions. Aside
supporting cells.
from pituicytes, the only other cellular elements
'in the neurohypophysis are the endothelial
The unmyelinated axons of the cells and the blood cells in the sinusoids.
parvocellular secretory neurons are relatively
short. They terminate in the 'median
eminence. The axonal terminations are where
the hypophysiotropic hormones that are
synthesized by the cells are stored. The axons,
together with the numerous capillaries and
supporting cells that are associated with them
comprise the median eminence.
The unmyelinated axons of the
magnocellular secretory neurons, on the
other hand, are much longer than those of
the parvocellular secretory neurons. They
originate from the nuclei (i.e., supraoptic and
paraventricular) where the cell bodies are
located, pass through the pituitary stalk and end
in the posterior lobe of the pituitary gland.
In the pituitary stalk, the proximal segments
Fig. XVIII-4. Pituitary Gland, Posterior Lobe.
of the axons comprise the hypothalamo-
Most of the cells are supporting cells called
hypophyseal tract, the main component of the pituicytes. Among the cellular and fibrillar
pituitary stalk. In the posterior lobe, the distal elements are deeply staining aggregations of
segments of the axons comprise the bulk of the secretory granules called Herring bodies (Hb)
lobe. and numerous blood vessels (bv). H&E x400.

[5TEB/\\: fA GOf~Z/\LES'TEXTBOOK OF HISTOLOGY

 In brief, the neurohypophysis is simply the Histologically, the parenchyma of the
downward continuation of the hypothalamus. It anterior lobe consists of epithelial cells, most of
consists of: a) axons of the secretory neurons which are secretory in nature. The parenchymal
whose cell bodies are in the hypothalamus, b) cells form irregularly-arranged anastomosing
pituicytes, and c) sinusoids. cords and clusters that are surrounded by
The neurohypophysis is not a gland. It fenestrated sinusoids. A minimal amount of
does not produce, but only stores and releases reticular tissue exists between the epithelial
(secretes) hormones. cells and the sinusoids. It serves as the
supporting structure (stroma) for the cell cords
and sinusoids.
Adenohypophysis
The adenohypophysis is the part of the Parenchymal Cells of the Auterior Lobe
pituitary gland that produces hormones. of the Pituitary Gland
Whereas the neurohypophysis is more fibrous In LM preparations that use a set of acid
than cellular, the adenohypophysis is more dyes, the parenchymal cells in the anterior lobe
cellular than fibrous. The adenohypophysis has of the pituitary gland can be classified into two
three regions: a) anterior lobe (pars distalis), generaltypes-chromophils, whose cytoplasm
b) pars tuberalis (pars infundibularis): and c) is intensely colored and chromophobes, whose
intermediate lobe (parsIntermedia). cytoplasm is pale staining. The chromophobes
The pars tub era lis forms a thin and comprise about 65% while the chromophils
incomplete sleeve around the pituitary stalk of comprise 35%of the parenchymal cells.
the neurohypophysis. Itis inferiorly continuous The chromophils are further classifiedinto
with the anterior and intermediate lobes. 1) acidophils (alpha cells) and 2) basophils
" (beta cells). The cytoplasm of acidophils stains
Anterior Lobe (Pars Distalis) red with acid dyes while that of the basophils
The anterior lobe comprises about 70% of stains blue or purple. In addition, the nucleus of
the pituitary gland. Nearly all the hormones the basophils is less dense, and is thus, lighter-
produced by the pituitary gland come from this staining than that of the acidophils.
lobe. The staining characteristics of the
chromophobes and chromophils in LM
preparations have nothing to do with true
acidophilia or basophilia because all the dyes
used are acid dyes. Furthermore, the dyes are
taken up mostly by the secretory granules of the
cells and not by the organelles.

Fig. XVIII-S. Pituitary Gland, Anterior Lobe.
The three types of epithelial cells that populate
the anterior lobe of the pituitary gland are
demonstrated in the photomicrograph. They are
acidophils (a), basophils (b), and chromophobes
(c). Note that the cords of epithelial cells are
surrounded by sinusoids (5). H&E x400.
Chrornophils
Electron microscopy and histologic
studies utilizing special techniques have shown
that there are actually five-not just two-
types of chromophils. The classification of
the chromophils into five types is based on
the hormone that the cells secrete and their
appearance in electron micrographs. The cell
types,whichhavebeen named afterthe hormones
they produce, are the: 1) somatotrophs (STH

ENDOCRINE SYSTEM Cd
cells), 2) mammotrophs (lactotrophs),
3) thyrotrophs, 4) corticotrophs, and S)
gonadotrophs. The somatotrophs and the
mammotrophs are the acidophils while the
thyrotrophs, corticotrophs, and gonadotrophs
are th~ basophils of traditional light microscopy.
Acidophils
Somatotrophs and mammotrophs look alike
in routine LM histologic preparations. They all
possess a reddish yellow-staining cytoplasm,
a dense nucleus, and distinct cell outline. But
the two cell types can be distinguished from
each other in electron micrographs-the
mammotroph has larger cytoplasmic secretory
granules.
Somatotrophs comprise the majority
(about SO%)of the chromophils in the anterior
lobe. They secrete somatotropin (growth
hormone), one of the few hormones that do not
have specific target cells:'_it affects practically
all cells in the body.
Mammotrophs comprise about 20% of the
chromophils in the anterior lobe where they
are scattered singly. During pregnancy, they
increase in number and in size. They secrete
the hormone prolactin (mammotropin) that
stimulates growth and activity of the mammary
glands during pregnancy and lactation. In non-
pregnant women, the role of prolactin is not
clear yet. In men, prolactin probably plays a role
in regulating testicular function.
Most acidophils secrete only one type of
hormone, but a few,the somatomammotrophs,
secrete both somatotropin and prolactin.
Basophils
Basophils are less numerous but are larger
than acidophils, The three cell types that
comprise basophils cannot be distinguished
from each other under the LM using acid dyes or
H&E stains, but their morphological differences
are evident in EMs.
Thyrotrophs comprise about S% of the
chromophils in the anterior lobe. They are
polygonal cells that are provided with long
cytoplasmic processes. Their secretor~
[STEB;\\: & GO\:z/\L.ES'TeXTBOOK OF HISTOLOGY
granules, which are concentrated close to the
cell membrane, are the smallest among all the
secretory cells of the anterior lobe. Thyrotrophs
produce thyrotropin (thyroid-stimulating
hormone, TSH) which stimulates the cells
in the thyroid gland that are responsible for
producing thyroid hormones.
Corticotrophs are ovoid cells that
constitute 20% of the chromophils in the
anterior lobe. Their secretory granules are
relatively few and are just slightly larger than
those of thyrotrophs. Corticotrophs secrete
corticotropin (adrenocorticotropin,
adrenocorticotropic hormone, ACTH) which
stimulates certain cells in the adrenal cortex to
produce hormones.
Gonadotrophs are large, round cells that
constitute around S% of the chromophils in
the anterior lobe. They are distributed singly
throughout the anterior lobe. Their secretory
granules are moderate and variable in size.
Gonadotrophs produce gonadotropins,
~..e., luteinizing hormone (LH) and follicle
stimulating hormone (FSH), whose functions
have been discussed in the chapters on the
male and female reproductive systems. It is
not known yet whether there are two types of
gonadotrophs (one producing LH and the other
producing FSH) or just one type that produces
both hormones.
Chromophobes
Electron microscopy and histologic studies
utilizing special techniques have shown that
most chromophobes are actually chromophils
(mostly corticotrophs), but they have pale
cytoplasm in routine LM preparations because
they are resting, or have just recently released
most of their secretory granules, or are still in
the process of producing new ones. In other
words, the few secretory granules they possess
are inadequate to impart enough cytoplasmic
coloration to distinguish them as chromophils
under the LM with the use of traditional acid
dyes.
 There are, however, two kinds of
chromophobes that are distinct from
chromophils. One type, the folliculostellate
cell (FS cell), has long, branching processes. It
isnon-secretory and performs some supportive
role for the other cells. The second type is an
undifferentiated stem cell.

Pars Tuberalis (Pars Infundibularis)

The tubular sleeve that the pars tuberalis
forms around the pituitary stalk is not uniform
in thickness, it ranges in thickness from 2 to
60 lAmand is thickest anteriorly. The pars Fig. XVIII-6. Pituitary Gland, Intermediate
tuberalis i\ separated from the pituitary stalk Lobe. The intermediate lobe of the pituitary
by connective tissue that is continuous with the , gland is between the anterior and posterior
lobes. In this specimen, most of the intermediate
pia-arachnoid membrane of the brain.
The pars tuberalis is more vascular than
the anterior lobe because the blood vessels that
comprise the hypophyseoportal system (see
page 288) traverse it. Its parenchymal cells are
mostly gonadotrophs and thyrotrophs that are
arranged parallel to the blood vessels in cords,
clusters, and sometimes, in the form of follicles
that contain colloid.
Intermediate Lobe (Pars Intermedia)
The intermediate lobe of the pituitary
gland is the thin and poorly developed region
of the gland that is in between the posterior
lobe and the anterior lobe. Its boundary with
the anterior lobe is demarcated by a groove that
represents the 6riginallumen ofRathke's pouch.
The intermediate lobe is better developed in
the fetus than in adults, where it is rudimentary.
In routine histologic sections, it is easy to
recognize because of the presence of follicles
(Rathke's cysts) that contain some eosinophilic
colloidal material.
The parenchymal cells of the intermediate
lobe form irregular clusters. They contain
some fine secretory granules. They synthesize
melanocyte-stimulating hormone (MSH)
whose production in adults is minimal to nil.
The physiologic action of MSH in humans
is unknown. In addition to MSH, evidence
lobe is occupied by follicles called Rathke's cysts,
H&E x100.
indicates that the cells of the intermediate lobe
also produce fJ endorphins.
Hypothalamic Control over the
Anterior Lobe of the Pituitary
Gland
As discussed in the preceding
paragraphs, the anterior lobe of the pituitary
gland secretes six important hormones: 1)
somatotropin, 2) prolactin, 3) thyrotropin,
4) corticotropin,S) FSH, and 6) LH. The
secretion of these hormones is mainly
regulated by the hypothalamus through its
hypophysiotropic hormones.
As mentioned earlier in this chapter,
the cell bodies of the secretory neurons that
produce the hypophysiotropic hormones are
widely distributed in the hypothalamus, but
their axonal terminations are in the median
eminence. The hypophysiotropic hormones,
after they are produced in the cell body of the
neurons, are stored in the terminations of the
axons in the median eminence. When needed,
the hormones are secreted into the capillaries
and transported by the blood to their target cells
in the anterior lobe of the pituitary gland via the
hypophyseoportal system.

ENDOCRINE SYSTEM €fI


Fig. XVIII-7. Pineal Gland. In the pineal gland,
most of the cells are pinealocytes. Among the
cells-which are arranged in cords or clusters
around in between capillaries-are calcified
concentric lamellar bodies called brain sands
(bs). H&E x1 00.
The hypophyse oporta] system refers
to the plexus of veins that travels from the
median eminence to the anterior lobe of the
pituitary gland. This venous plexus is a portal
system because it receives blood from a set
of capillaries and drains into another set of
capillaries. Specifically,the veins comprising the
hypophyseoportal system receive blood from
the capillaries that supply the median eminence,
and after traveling downward along the pars
tuberalis, drain into another set of sinusoidal
capillaries that supply the anterior and posterior
lobes of the pituitary gland.
As a rule, each hypophysiotropic hormone
exerts control over more than one anterior
pituitary cell type. As examples, TRH stimulates
production of both thyrotropin and prola~tin
while GnRH stimulates the secretion of lfuth
FSHandLH.
PINEAL GLAND (EPIPHYSIS
CEREBRI; PINEAL BODY)
The pineal gland is a small (8 mm long;
about 120 mg in weight) conical structure
whose base is attached to the posterior portion
of the roof of the third ventricle of the brain by
two short stalks.
ESTE8!\N & GCNZl\LES' TEXTBOOK OF HISTOLOGY
Development of the Pineal Gland
The pineal gland arises in the embryo
as a hollow evagination of the roof of the
diencephalon. As it develops, its cavity getsfilled
with cells and eventually the gland transforms
into a solid organ. The pineal gland is well-
developed in children. At puberty, it starts to
involute.
Histology of the Pineal Gland
The pineal gland is enveloped by a thin
connective tissue capsule that is derived from
pia mater. Connective tissue septa arise from
this capsule and incompletely divide the gland
into irregular lobules. The tissue septa also
serve as passageways for blood vessels and
unmyelinated nerve fibers.
The parenchymal cells that fill the lobules
of the pineal gland are mostly pinealocytes.
Pinealocytes are modified neurons that
comprise 95%of the cell population of the gland.
In the lobules, they are arranged in short cords
orin clusters that are surrounded by fenestrated
capillaries. In routine histologic preparations,
pinealocytes have poorly defined borders and
basophilic cytoplasm. Their large nucleus,
often irregular or even lobated, has a prominent
nucleolus. Their branching processes, which
are not easily appreciated in H&E preparations,
terminate in expansions near the fenestrated
capillaries.
Dispersed among the pinealocytes are
supporting cells that are called interstitial
cells. The interstitial cells are smaller than
pinealocytes. They are morphologically similar
to astrocytes.
Aside from pinealocytes and interstitial
cells, mast cells are also often seen in the pineal
gland, a fact that may explain the organ's high
histamine content.
One striking histologic feature of the pineal
gland is the presence of extracellular bodies
called brain sands (acervuli, psammoma
bodies, or corpora arenacea) within the
lobules. These are calcified bodies that have
a concentric lamellar structure. They tend
to increase in number with age. They are
radiopaque. This is why the pineal gland is
visible and is sometimes used as a landmark in
X-ray studies of the brain.
As one grows older, the interstitial cells
increase in number and hyalinization of parts
of the pineal gland occurs.
Hormone Produced by the Pineal
Gland
The pineal gland produces the hormone
m~J.atonin whose secretion is stimulated by
darknes and inhibited by light.
Melatonin is synthesized by the
pi)l,ea.locytes.It is a ve~y important hormone
in animals that breed seasonally because it
regulates the animals' sexual develop men
reproductive cycle (seasonal breeding),
hibernation, and other metabolic processe .
In humans, however, the physiologic
function of melatonin is not yet known. There
are speculations that it affects circadian
Fig. XVIII-B. Thyroid and Parathyroid Glands. The thyroid gland which is located on the anterior
neck consists of two lateral lobes connected at the midline by the isthmus. The parathyroid glands,
on the other hand, usually consist of two pairs attached to the posterior surface of the lateral lobes
of the thyroid gland.
rhythms and sleep patterns. It is associated in
the development of seasonal affective disordef
a condition that is characterized by depressio
during winter in countries where there is
one. There is also speculation that it inhibits
sexual development such that children mature
sexually only when their melatonin levels
drop significantly during pubert At present,
however, all these are just speculations. But it is
known that in humans, the normal melatonin
blood levelhas a diurnal pattern (i.e., it is higher
at night than during the da -); and ismuch higher
in children than in adult .
THYROID GLAND
The thyroid gland is the largest of the
endocrine glands. It is 25 to 40 g in weight and
is slightly bigger in women than in men. It is an
unpaired gland that lies on the anterior aspect
of the neck. Grossly, it consists of two lateral
lobes (right and left) and an isthmus.
The lateral lobes are broad inferiorly but
tapered superiorly. They adhere to the lateral
(one on each side) aspects of the pharynx,
larynx, esophagus, and trachea. Their inferior
parathYWI
glandS
{behind
thyroid)
ENDOCRINE SYSTEM _
borders are at the level of the sixth tracheal
cartilage. The lateral lobes are connected to each
other by a narrow horizontal bridge of glandular
tissue, the isthmus, which extends across the
trachea .usually in front of the second and third
tracheal cartilages. In some individuals, the
thyroid gland also has a small pyramidal lobe,
which consists of glandular tissue that extends
upward from the isthmus.
Fig. XVIII-9. Thyroid Gland. The gland is
enveloped by a capsule (ca) that sends septa (s)
into the substance of the organ to divide it into
lobules (lo). The lobules are occupied by follicles
(fo) which are cystic structures that contain a gel-
like material called colloid (co). H&E x100.
Fig. XVIII-10. Thyroid Follicles. The thyroid
follicles (fo) which occupy the thyroid lobules
are ball-like structures whose wall is formed by
a simple epithelium and whose cavity is filled
with colloid (co). The epithelium has two types
of cells: follicular cells (fc) which comprise 90%
of the epithelial cells, and the bigger and lighter
parafollicular cells (pc) which are scattered singly
or in clusters among the follicular cells. Thyroid
Gland, H&E x100.
ESTEBAN & CC)NZALES' TEXTBOOK OF HISTOLOGY
Development of the Thyroid
Gland
The thyroid gland arises during early
embryonic life as an epithelial invagination at
the base of the tongue. It migrates downwards
and reaches its final position in the lower part
of the neck in the 7th week of intrauterine
life. It remains connected for a time to the
base of the tongue by the thyroglossal duct,
which later disappears. The point of origin of
the thyroid gland is indicated by the foramen
cecum,the apex of the V-shaped furrow (sulcus
terminalis) that separates the anterior from the
posterior tongue.
Histology of the Thyroid Gland
The thyroid gland is enclosed by two
capsules, an outer capsule that is derived from
the pretracheallayer of the deep cervical fascia
and a true capsule that closely invests the gland
and sends connective tissue septa into the organ
to form poorly-defined lobules. The connective
septa also serve as pathways for blood and
lymphatic vessels and nerves. The connective
tissue within the thyroid gland is infiltrated by
lymphocytes and small lymphoid nodules are
occasionally seen.
Each thyroid lobule is filled with irregular
spherical, cystic structures called follicles
(thyroid follicles) that are surrounded by
fenestrated capillaries and supported by a thin
connective tissue stroma-consisting primarily
of reticular fibers and cells-that is continuous
with the septa.
The follicles are 0.02 to 0.9 mm in diameter.
Their cavity is occupied by a homogenous, gel-
like material known as colloid while their wall
is formed by a simple epithelium.
Two types of cells comprise the simple
epithelium that forms the wall of thyroid
follicles: follicular cells (principal cells)
and parafollicular cells (mitochondria rich
cells; C cells, clear cells). The shape of the
epithelial cells and the staining property of the
colloid depend on the functional state of the
follicle. In inactive follicles, the epithelial cells
are squamous or cuboidal and the colloid is
acidophilic. In active follicles, the epithelial cells
are tall cuboidal or columnar and the colloid is
basophilic.
Follicular cells make up the overwhelming
majority of the epithelial cells in the thyroid
follicle. In routine histologic preparations, they
are seen as having a round nucleus that contains
fine chromatin material and one or two nucleoli,
and a slightly basophilic cytoplasm. The
luminal surface of follicular cells is provided
with numerous microvilli, but these are not
discernible under the LM.
Parafollicular cells comprise only
0.1 % of the epithelial cell population of the
thyroid follicle. They are scattered singly or
in small groups in the epithelium. For a time,
histologists thought that parafollicular cells
are also present in between thyroid follicles
because in LM preparations, many of them
are found in the interstices between follicles,
but the electron microscope has shown that'
all parafollicular cells actually form part of the
follicular epithelium. The parafo~~icular cells
rest on the basal lamina of the epithelium but
unlike follicular cells, their apices do not touch
the colloid. In routine histologic preparations,
they are easy to identify because they are much
bigger and lighter-staining than follicular cells.
Their cytoplasm contains numerous small
secretory granules.
Thyroid Hormones
The hormones produced by the thyroid
gland are thyroxine (T4) and triiodothyronine
(T3), which are collectively referred to as
thyroid hormones, and calcitonin.
The production and secretion ofthe thyroid
hormones (T3 and T4) are functions of the
follicular cells but the follicular cells do not
directly synthesize the hormones. Instead, they
synthesize thyroglobulin, a large glycoprotein,
which they discharge by exocytosis into the
colloid. They likewise collect iodine from
the blood in the capillaries that surround the
follicles and transport it to the lumen of the
follicle.
In the colloid,some ofthe tyrosine residues
present in the thyroglobulin are iodinated and
condensed to form the, two principal thyroid
hormones, thyroxine (T4) and triiodothyronine
(T3), with the aid of the enzyme thyroid
peroxidase.
T3 and T4 remain bound to thyroglobulin
until they are secreted, during which time the
follicular cells ingest colloid by endocytosis,
hydrolyze the peptide bonds that bind the
thyroid hormones to the thyroglobulin,
and release the thyroid hormones into the
capillaries.
The thyroid hormones regulate the
metabolism ofproteins, carbohydrates, fats, and
some vitamins.
Control of T3 and T4 secretion is the
function of thyrotropin (TSH; thyroid
stimulating hormone).
Calcitonin (thyrocalcitonin), on the
other hand, is synthesized and secreted by the
parafollicular cells. It lowersblood calciumlevel
by suppressing the bone resorption activity of
the osteoclasts. Incidentally, calcitonin is
probably also secreted by other still unidentified
cells of the body because it is present in the
blood of people who have had their thyroid
gland surgically removed.
The secretory activity of the parafollicular
cells is regulated directly by blood calcium
level. A high blood calcium level stimulates ,.
while a low blood calcium level suppresses
secretion of calcitonin by the parafollicular
cells.
PARATHYROID GLAND
The parathyroid glands, usually two
pairs (superior and inferior), are yellowish-
brown, tiny, ovoid bodies that are attached to
the posterior surface of the lateral lobes of the
ENDOCRINE SYSTEM Wi

thyroid gland. Each parathyroid gland is about
6 mm long, 3 to 4 mm wide and 1 to 2 mm thick,
and about SO mg in weight.
Development of the Parathyroid
Gland
The inferior parathyroid glands arise in
the embryo from the third pharyngeal pouch
together with the thymus. When the thymus
descends into the thoracic cavity, it pulls the
inferior parathyroid glands with it, but the latter
do not descend all the way to the thorax. They
come to rest on the posterior surface of the
thyroid gland. Occasionally though, parathyroid
glands are found in the company of the thymus
in the thoracic cavity. The superior parathyroid
glands, on the other hand, arise from the fourth
pharyngeal pouch and attach themselves to the
thyroid glands as the latter migrates downward
to the inferior portion of the neck.
Histology of the Parathyroid
Gland
The parathyroid glands lie within the
capsule of the thyroid gland. In addition, deep
to the thyroid capsule, each parathyroid gland
D' ESTEBi\N& GONZ!\LES' TEXTBOOK OF HISTOLOGY
Fig. XVIII-11. Parathyroid Gland.
The parenchyma of the gland is
made up of two types of cells: chief
cells which are unlabeled in the
photomicrograph but comprise
most of the cells, and the bigger
and more eosinophilic oxyphil
cells (0) that are scattered singly
or in small groups among the
chief cells. The parenchymal cells
are arranged in cords or clusters
surrounded by fE\Q_estrated
capillaries (c). H&E x400.
is enveloped by its own thin connective tissue
capsule. From this latter capsule, connective
tissue septa arise and divide the gland into
poorly-defined lobules. In the lobules, the
stroma is formed by delicate connective tissue
that consists mainly of reticular fibers and cells.
The parenchyma of the parathyroid gland
consists of epithelial cells that are arranged
in cords and clusters that are surrounded by
fenestrated capillaries.
Adipose cells are also relatively abundant
within the lobules of the parathyroid gland
where they intermix with the parenchymal cells.
By the age of2S years, they comprise about 30%
of the volume of the gland. The adipose cells
increase in number with age.
Follicles are occasionally present in
the parathyroid glands, especially in older
individuals. These follicles resemble those
found in the thyroid gland.
Parenchymal Cells of the Parathyroid
Gland
Two types of parenchymal cells exist in the
parathyroid gland: chief cells (principal cells)
and oxyphil cells (acidophil cells).
 The chief cells comprise majority of the tubules; increasing the conversion of vitamin
parenchymal cells. A chief cell is a relatively D to its active form; increasing the excretion of
small (8 to 10 mm in diameter) polyhedral cell. phosphate by the kidneys; and promoting the
In routine histologic preparations, its nucleus absorption of calcium in the digestive tract.
is prominent, rather large, and vesicular. Its High level of blood calcium, on the other
homogenous cytoplasm is faintly eosinophilic hand, suppresses the activity of the chief cells,
and it contains small, secretory granules that in causing calcium to be deposited in bone.
electron micrographs, are seen to be membrane-
bound.
ADRENAL GLAND
The oxyphil cells are absent in children. (SUPRARE:NALGLAND)
They appear shortly before puberty and
increase in number with age. They occur singly The paired adrenal glands (left and right)
or in clusters in the lobules. An oxyphil cell is are flat, pyramidal organs that rest on the upper
bigger than a chief cell but its nucleus is slightly pole of each kidney. Each gland weighs about
smaller. Its cytoplasm is intensely eosinophilic 5 g and is 50 mm long or high, 30 mm wide,
because of the presence of many acidophilic and 10 mm thick. The adrenal gland presents
granules. Electron micrographs show that an indentation (hilus) at the middle of its
these granules are actually mitochondria anteromedial aspect; it is here where the adrenal
with numerous cristae. Oxyphil cells are non- vein leavesthe gland. The adrenal arteries enter
secretory and their function is unknown. the gland through its capsule.
They are suspected to be chief cells that are in The stroma of the adrenal gland consists
a different physiological state. This assertion of a relatively thick connective capsule and
is strengthened by the fact that there are cells elements from this capsule (mostly reticular
in the parathyroid gland (transitional cells) fibers and cells) that penetrate the gland all the
that have structural characteristics that are i~' way to the medulla. The parenchyma of the
between the chief cells and oxyphil cells. gland, on the other hand, consists of epithelial
cells that are mostly secretory in nature.
Parathyroid Hormone (PTH; An examination of a coronal section of the
Parathormone) adrenal gland shows that the gland consists of
two regions: cortex and medulla.
The parathyroid gland secretes only one
hormone, the parathyroid hormone, which is
synthesized and secreted by the chief cells. adrenal
gland
The parathyroid hormone is the most
important regulator ofblood calcium level and
its secretion is controlled by the level ofionized
calcium in the blood.
When the blood calcium level is low, the
chief cells are stimulated to produce and
secrete parathyroid hormone, which increases
the circulating blood level of calcium (i.e.,
through the osteoblasts, see chapter on
cartilage and bone) by indirectly increasing the
number and activity of osteoclasts; inhibiting
the bone formation activity of osteoblasts; Fig. XVIII-12. Adrenal Gland. The adrenal gland
enhancing calcium reabsorption in the renal rests on the superior pole of the kidney.

ENDOCRINE SYSTEM +a

Fig. XVIII-13. Adrenal Gland. This scanning
photomicrograph of the adrenal gland shows
the capsule that envelops the organ and the
two regions that comprise the gland: cortex and
medulla. H&E x40.
The cortex lies immediately beneath the
capsule. It completely surrounds the medulla,
which occupies the inner area of the gland.
The adrenal gland can actually be considered
as consisting of two separate endocrine.glands
because the cortex differs from the medulla
embryologically, structurally, and functionally.
,.
Development of the Adrenal
Gland
The cortex of the adrenal gland is
mesodermal in origin while the medulla is
ectodermal.
The cortex is derived from mesothelial
cells of the peritoneum that are adjacent to the
developing kidneys. These cells differentiate to
form the primitive adrenal cortex. Later, another
group of mesothelial cells, which are smaller
than those in the primitive adrenal cortex,
invades the area of the developing adrenal gland
and surrounds the primitive adrenal cortex.
These cells comprise the definitive adrenal
cortex. During the first postnatal month of Iife,
the primitive adrenal cortex rapidly regresses
and its cells replaced by the cells of the definitive
cortex, but only at puberty is the structure of the
adult adrenal cortex achieved.
ESTE8;.\N& GONZALES' TEXTBOOK OF HISTOLOGY
The medulla, on the other hand, is·
derived from neural crest cells that form the
sympathetic system. These cells migrate to,
and invade the medial aspect of the developing
adrenal cortex where they later get entrapped by
the cortical cells.
Adrenal Cortex
The cortex comprises 80% to 90% of the
adrenal gland. It is essential to life. Its function
is to produce steroid hormones, collectively
referred to as adrenocortical hormones,which
play vital roles in the body's metabolic activities.
The adrenocortical hormones
are categorized into three classes (i.e.,
mineralocorticoids, glucocorticoids, and
androgens). Many hormones comprise each
class, but most are secreted in physiologically
insignificant amounts. The only physiologically
important adrenocortical hormones are
the m iner alocor tico id aldosterone, the
glucocorticoids cortisol and corticosterone,
and the androgens dehydroepiandrosterone
(nHEA) and androstenedione.
The adrenal cortex, when seen under LM, -I
consists of three concentric zones or layers that
are distinguishable by the arrangement and
appearance of their parenchymal cells. From
the outside going inwards, these layers are the a)
zona glomerulosa, b) zona fasciculata, and, c)
zona reticularis.
Fig. XVIII-14. Adrenal Cortex. Immediately
deep in the capsule (c) of the adrenal gland is
the cortex. It hasthree regions: zona glomerulosa
(zg), zona fasciculata (zf), and zona reticularis (zr).
H&E x100.
Zona Glomerulosa
The zona glomerulosa occupies the
outermost region and makes up about 10% to
15%of the adrenal cortex. In routine histologic
preparations, the zone is seen to consist of
rounded or pyramidal epithelial cells that are
arranged in irregular ovoid clusters that are
separated by sinusoids. The parenchymal cells
have a deeply-staining nucleus that contains one
or two nucleoli. Their cytoplasm is scanty and
eosinophilic.
The parenchymal cells of the zona
glomerulosa produce mineralocorticoids,
mainly aldosterone whose primary effect is Fig. XVIII-1S. Zona Fasciculata. This high-
magnification photomicrograph shows the
to increase Na' reabsorption by the distal and parenchymal cells of the zona fasciculata. The
collecting tubules of the kidney. cells are called spongiocytes because their
cytoplasm is vacuolated. The spongiocytes-
The activity of the parenchymal cells that unlabeled in the section-are arranged in thin
produce aldosterone is -regulated primarily cords perpendicular to the capsule. The cords
by the renin-angiotensin system of the are separated by sinusoidal capillaries (c) that
kidneys and only secondarily by corticotropin are lined by endothelial cells (e). Adrenal Gland,
H&E x400.
(ACTH) that comes from the anterior lobe of
the pituitary gland.
The primary regulator of the activity of the
cells of the zona fasciculata is corticotropin
Zona Fasciculata
(ACTH).
The zona fasciculata forms the thickest layer
and comprises about 75% to 80% of the adrenal Zona Reticularis
cortex. In routine histologic preparations, the
The zona reticularis is the innermost and
zone consists of parenchymal cells that form
thinnest layer of the adrenal cortex. It accounts
long, thin (usually one-cell thick), straight cords
for only 5% to 10% of the volume of the adrenal
that are arranged perpendicular to the capsule.
cortex. Its parenchymal cells, which are small
The cords are separated by sinusoids. The
compared to those in the outer two zones, are
parenchymal cells are large and polyhedral in
arranged in short anastomosing cords that are
shape and their nucleus is vesicular and contains separated by sinusoids. The parenchymal cells
a prominent nucleolus. They have abundant, are histologically similar to the cells of the
faintly-acidophilic cytoplasm that contains zona fasciculata but they are less vacuolated
many empty spaces that represent lipid droplets because they have less lipid droplets and their
extracted by reagents. The empty spaces give nucleus stains more intensely. Practically no
the cells a vacuolated appearance that is why the mitosis occurs among the parenchymal cells in
cells are sometimes called spongiocytes. this zone. Some histologists consider the zona
The parenchymal cells of the zona reticularis as a "graveyard" where old cells from
fasciculata produce glucocorticoids, the two other zones eventually go.
mainly cortisol and corticosterone, which The parenchymal cells of the zona
regulate carbohydrate, lipid, and protein reticularis produce a small quantity of
metabolism. glucocorticoids (cortisoland corticosterone)

ENDOCRINE SYSTEM +&

and the androgens dehydroepiandrosterone
(DHEA) and androstenedione. Androgens
usually have masculinizing effects and they
promote protein anabolism and growth. But
the potent androgens are those secreted by the
testes. The adrenal androgens have less than
20% the activity of testicular androgens and they
hardly have any physiological effect on normal
people. In other words, the amount of androgen
that is normally secreted by the adrenals, which
is equal in both men and women, is not enough
to induce masculinization in women.
The activity of the parenchymal cells of the
zona reticularis is regulated by corticotropin
(ACTH). Gonadotropins do not affect them.
Fig. XVIII-16. Adrenal Medulla. The parenchymal
cells (p) of the adrenal medulla, called
phaeochromocytes, are arranged in clusters or
in cords that are surrounded by sinusoids (5). H&E
x400.
Adrenal Medulla
The medulla comprises 10% to 20% of the
adrenal gland. Its central area contains large
veins (medullary veins) that drain the entire
gland. Unlike the adrenal cortex, the adrenal
medulla is not essential to life, but its hormones
help the individual cope with emergencies.
The parenchymal cells of the medulla
(phaeochromocytes, chromaffin cells) are
arranged in groups or in thick cords that are
ESTEBANs (iCNz/\LES' TEXTBOOK OF HISTOLOGY
surrounded by sinusoids and richly supplied'
with myelinated nerves-in fact, all the
parenchymal cells are associated with endings
of preganglionic sympathetic neuron. They
are polyhedral cells with basophilic cytoplasm
that, when compared to the cortical cells in
routine histologic preparat~ons, have a larger
and more darkly-staining nucleus. They are
called chromaffin cells because the secretory
granules in their basophilic cytoplasm exhibit
chromaffin reaction, i.e., when treated
with oxidizing agents like chromate, the
cathecolamines that they contain get oxidized
and turn brown.
Scattered among the chromaffin cells are
sympathetic neurons (ganglion cells) which
are the sources of the myelinated nerves that are
associated with the chromaffin cells. Ganglion
cells are large cells that are round or polygonal
with prominent nuclei.
Adrenal Medullary Hormones
The hormones produced by the
adrenal medulla are collectively referred
to as catecholamines. They are secreted
by the chromaffin cells and include
epinephrine (adrenaline), norepinephrine
(noradrenaline), and dopamine, which,
incidentally, is also synthesized by nervous
tissue.
About 90% of chromaffin cells secrete
epinephrine and about 10% secrete
norepinephrine while the cells that secrete
dopamine have not been identified yet. The
chromaffin cells store their secretion in their
cytoplasm in the form of granules. Cells that
secrete and store norepinephrine exhibit
a stronger chromaffin reaction and their
secretory granules are more electron-dense
than those of the epinephrine-secreting and
-storing cells.
The cathecolamines have no specific target
organs or cells. Their effects are widespread and
involve practically all the cells of the body and
are designed to enable the individual to cope
with "fight-or-flight" situations, e.g., increase
in heart rate, blood pressure, and blood glucose
level.
Secretion of the cathecolamines is
controlled by the preganglionic neurons of the
sympathetic nervous system. The chromaffin
cells are thus essentially sympathetic
ganglion cells that respond to stimulation
by the preganglionic neurons not by sending
nervous impulses but by releasing hormones
by exocytosis and sending these chemical
messengers via the bloodstream to effector
organs and cells.
In addition to catecholamines, the
chromaffin cells also synthesize a wide variety
ofbioactive amines and peptides.
Paraganglia
The term paraganglia refers to small
clusters of chromaffin cells that are found
outside the adrenal medulla. They are Histology of the Islets of
associated with autonomic ganglia, nerves
of the sympathetic nervous system, and the
aorta. The chromaffin cells of the paraganglia' .
secrete mainly norepinephrine.
ISLETS OF LANGERHANS
(PANCREATIC ISLETS)
As discussed in the chapter on the accessory
glands of the digestive system, the pancreas is
both an exocrine and an endocrine gland. Its
endocrine part is composed of the islets of
Langerhans.
Development of the Pancreas
The exocrine and endocrine portions of the
pancreas have the same embryonic origin-the
endoderm that lines the duodenum. In the 3rd
month of intrauterine life, the cells destined
to form the islets migrate away from the
pancreatic ducts, aggregate around capillaries,
and differentiate into islet cells. During the
5th month of fetal life, the cells start producing
hormones.
Langerhans
The islets of Langerhans are 100 to 200
~m in diameter each. In routine histologic
Fig. XVIII-17. Pancreas. The
photomicrograph shows
several islets of Langerhans
(at arrows) surrounded by the
darker-staining pancreatic
acini. H&E x100.
ENDOCRINE SYSTEM +8
Fig. XVIII-18. Islet of
Langerhans. Compare the
cells of the islet (ic) to those
of the pancreatic acini (ac).
Note that the islet cells are
smaller and paler-staining
than the acinar cells. The cells
of the islets of Langerhans
form a compact mass
supplied with numerous-
capillaries (c). Pancreas, H&E
x400.
preparations, they are seen as consisting of
small aggregations of pale-staining cells that are
scattered among the darker-staining cells that
belong to the exocrine portion of the pancreas.
Each islet consists of 2,000 to 3,000 cells that
form a compact mass that is criss-crossed by
fenestrated capillaries and surrounded by a thin
layer of reticular tissue.
There are over a million islets of Langerhans
in the pancreas, but they account for only 2%
of the volume of the organ. They are more
numerous in the tail than in the body or head
of the organ.
Cells of the Islets of Langerhans
The cells of the islets of Langerhans are
polygonal and are typically polarized toward
the capillaries into which they discharge
their secretions. They consist of four distinct
cells types: a-, ~-, B-, and F-cells. Each cell
type secretes a hormone. The cell types are
not distinguishable in H&E preparations,
but they can be distinguished from each
ESTEBAN& GONZJ\LES'TEXTBOOK OF HISTOLOGY
other by employing special staining,
'immunocytochemistry techniques, and by
electron microscopy. In electron micrographs,
the main differentiating point between the cell
types lies in the structure of their secretory
granules.
The a-cells (A cells) comprise about 20%
of the islet cells. They are large cells and most
occupy the peripheral areas of the islet. They
have electron-dense secretory granules that are
uniform in size
The ~-cells (B cells) are the smallest but
most numerous, comprising 60% to 75%, of
the islet cells. Most are located in the central
area of the islets. Their secretory granules are
smaller and less electron-dense than those of
the a-cells.
The B-cells (D cells) are the largest of the
islet cells but they comprise only about 5%of the
islet cell population. They are scattered singly
all over the islets. Their secretory granules are
similar to those of the a-cells except that in
8-cells, the granules are less electron-dense.
 The F cells (pp cells) are rare. Like the Glucagon, which is also a polypeptide
8-cells, they are widely scattered and sometimes hormone, is elaborated and secreted by the
they occur among the pancreatic acini. Their a-cells. Like insulin, the effects of glucagon
secretory granules are irregularly shaped and of are varied and complex but its main action is to
variable electron density. increase blood glucose levelby ordering the cells
of the liver to produce glucose or release it from
Hormones Produced by the Islets the or~.m's glycogen store.
of Langerhans Secretion of glucagon by the a-cells is
regulated mainly by the glucose level ofblood-
The islets of Langerhans produce four (4) it is stimulated and inhibited by low and high
hormones: insulin, glucagon, somatostatin, blood glucose level respectively.
and pancreatic polypeptide.
Somatostatin is produced and secreted by
Insultn'Js secreted by the ~-cells. It is the 8-cells. Its main action is on the secretion
a polypeptide hormone that is involved in of the other cells of the islets. It inhibits the
numerous metabolic processes. Insulin affects secretion of glucagon, insulin, and pancreatic
practically all cells of the bod and its effects polypeptide. It also minimally diminishes the
are varied, very profound, and comple . Its most motility of the stomach, small intestine, and
notable action is facilitating entry of glucose, the gallbladder. Its release is stimulated by the
chief energy source of ceHs,into the cytoplasm increase in blood glucose that occurs after a
of cell. In addition toits effect on carbohydrate meal.
metabolism, it also affects protein and fa Somatostatin is also produced by some cells
metabolism, the activity of many enzyme , and of the digestive tract and the hypothalamus,
electrolyte transpor . but the physiologic effect of hypothalamic
The secretion of insulin by the ~-cells is' . somatostatin differs from that of pancreatic
regulated mainly by the level of glucose in the somatostatin.
blood. High blood glucose level stimulates its Pancreatic polypeptide'rs produced and
secretion and release. When insulin is released, it secreted by the F cells. It has been shown to
diffuses to cells throughout the body. Secretion slow down the absorption of food from the
and release of insulin by the ~-cells end when intestines, but its exact physiologic effect is still
the blood sugar level returns to normal. unknown. Its secretion is stimulated by low
Insulin secretion is also partly regulated by the blood glucose level, a meal containing protein,
autonomic nervous system and by some of the exercise, and fasting; it is also partly regulated
hormones secreted by the digestive tract e.g., by the autonomic nervous system.
enteroglucagon, secretin, cholecystokinin, The hormones of the islets of Langerhans
gastrin, and gastric inhibitory peptide (GIP)o affect each other. Somatostatin inhibits the
Inability of the islets of Langerhans to produce secretion of the three other hormones produced
enough insulin and/or the inability of the body by the islet; insulin inhibits the secretion of
cells to respond appropriately to insulin results glucagon; and glucagon stimulates the secretion
in a disorder called diabetes mellitus. of insulin and somatostatin.

ENDOCRINE SYSTEM ta
 Eye

he most complex sense organs of humans

T are the eyes (eyeballs).They are a pair of

globular structures located inside bony
cavities (orbital cavities: orbit) on the anterior
aspect of the skull. They are the only organs in
the body that contain sensory cells sensitive to
light waves and are, therefore, solely responsible
for the individual's sense of vision.
Each eye is about 2.4 em in its
anteroposterior (AP) diameter. Each occupies
only the anterior half of its orbital cavity:
the posterior half is occupied largely by fat
and muscles. The fat serves as a cushion that
protects the eye from injury during violent head Fig. XIX-1. Eye. Sagittal Section
movements. The muscles, on the other hand,
The eye is a sphere that consists of awall that
are collectively referred to as the extraocular
muscles (EOMs). They consist of six skeletal encloses a cavity. The wall has three histologic
layers while the cavity is partitioned into three
muscles (i.e., lateral rectus, medial rectus,
superior rectus, inferior rectus, superior unequal compartments (or chambers) that are
filled with transparent media.
oblique and inferior oblique) that originate
from the bony walls of the orbit and insert into The photosensitive cells (photoreceptor
the outer surface of the eyeball. The EOMs cells) of the eye are in the retina, the innermost
serve to direct the eyes to their object of of the three histologic layers that comprise the
concern. When the extraocular muscles of one wall of the organ. Light that enters the eye
eye contract, they do so in synchrony with those passes through several refractive media before
of the other eye. it reaches the retina. The refractive media
concentrate light and focus the image of the
The front aspect of the eye slightly
object being viewed on the photoreceptors in
bulges out of the anterior margin of the orbit.
the retina. The photo receptors then transmit
Nonetheless, the protruding portion is protected
the attributes of the image to the brain via the
by thin folds of tissue (upper and lower eyelids)
optic nerve for interpretation.
that can cover it when needed.
 anterior
chamber

Fig. XIX-2. Eye, Anterior Region. H&E x40.

Fig. XIX-3. Histologic Layers of the Eye. The

HISTOLOGIC LAYERS OF THE photomicrograph shows the histologic layers of
EYE the posterior wall of the eyeball. H&E x100.

From the most external to the most internal Sclera

(external means farther from while internal
means closer to the center of the eyeball), the
three histologic layers- (coats; tunics) that
comprise the wall of the eye are: 1) tunica
fibrosa (fibrous layer), 2) tunica vasculosa
(uvea; uveal tract), and 3) tunica interna
(retina). Of thethree layers, only the tunica
fibrosa envelopsthe eyeball completely;the two'
inner coats do not extend as far anteriorly as the
tunica fibrosa.
Tunica Fibrosa (Fibrous Layer)
The tunica fibrosa has two regions: 1)
sclera, and 2) cornea. The sclera comprises the
posterior S/6 while the cornea, the anterior 1/6 of
the tunica fibrosa. Grossly, the sclera is white
and opaque while the cornea is transparent. The
center of the corneal curvature is the anterior
pole, and the center of the scleral curvature is
the posterior pole of the eyeball.
The cornea and the sclera are continuous
with each other at the corneoscleral junction
(limbus) which is denoted by a shallow groove
internally and externally.
The sclera is a tough, dense irregular
connective tissue layer that is 0.3-1.0 ~m thick.
It is mainly made up of bundles of collagen
fibers (type I collagen) that are embedded in
moderate amount of ground substance. The
collagen bundles are arranged parallel to the
surface of the eyeball, but intersect in various
directions. Interspersed among the collagen
bundles are a few flattened fibroblasts.
Delicate mucous membrane (called
conjunctiva, see page 311) covers the exposed
surface of the scleraup to the edge ofthe cornea.
Conjunctiva also lines the inner surface of the
eyelids.
bulbar

Fig. XIX-4. Corneoscleral Junction and

Adjacent Structures. The corneoscleral junction
is otherwise known asthe limbus. Note the bulbar posterior
conjunctiva that covers the exposed portion of chamber
the sclera. H&E x100.

EYE_
 The aponeuroses of the extraocular muscles
insert into the sclera-the four recti 6-8 mm
posterior to the corneoscleral junction, and the
two obliques into the posterolateral quadrant.
External to the sclera, a sheath of fascia
(Tenon's capsule) that is made up of dense
connective tissue envelops the posterior portion
of the eyeball. Between the Tenon's capsule and
the sclera is a space (Tenon's space; episcleral
space) that contains a loose network of collagen
fibers (sometimes referred to as episclera).
These fibers connect the external surface of the
sclera to the Tenon's capsule.

Embedded in the sclera near the limbus

is a circumferential system of interconnecting
endothelium-lined and irregularly-shaped tubes,
collectively referred to as canal of Schlemm.
This canal serves as a drainage channel for
aqueous humor from the anterior chamber
to the venous system (further discussed later in
Fig. XIX-S. Cornea. The cornea has five
histologic layers: epithelium (ep), Bowman's
membrane (Bm), substantia propria (sp) that
consists of collagen fibers and fibroblasts called
keratocytes (ke), Descemet's membrane (Om),
and endothelium (en). H&E x400.

this chapter).
The optic nerve-which carries impulses
from the photoreceptorsof the eye to the brain
and which consists of the axons of the ganglion
cells of the retina (see page 306)-t;xits the
eyeball on its posterior surface. As soon as it
exits the eyeball, the optic nerve is enveloped
by a strong sheath of dura mater (dural sheath)
that blends with the sclera. The region where the
optic nerve exits the eyeball is called the optic
disc (optic papilla). Here, numerous small
holes that serve as passageways for bundles of
nerve fibers that form the optic nerve perforate
the sclera. This perforated area of the sclera is
called lamina cribrosa.
Cornea
The cornea is thicker than the sclera. It
is 0.55-1.0 mm thick. Its central region is
slightly thinner than its peripheral region. As
already mentioned, the cornea connects to the
sclera at the limbus. It is also connected to the
iris (page 305) at a point called the angle by
loose connective tissue fibers. These fibers
are collectively referred to as the trabecular
~~twork (trabecular meshwork).
The cornea is richly supplied with sensory
nerves but is avascular. Its cells derive nutrition
and oxygen partly from the blood vessels of
the highly vascular limbus and partly from the
aqueous humor that fills the anterior chamber
that is immediately behind it.
The cornea has five histologic layers. From
the most external to the most internal, these
are: 1) epithelium, 2) Bowman's membrane,
3) stroma (substantia propria), 4) Descemet's
membrane, and 5) endothelium.
The corneal epithelium is nonkeratinized
stratified squamous. It consists of 5 to 6 layers
of cells. The most superficial cells are provided
with microvilli that are soaked in a thin (about
7 ~m) protective layer of lipid and glycoprotein
(precorneal tear film). The cellsin the deepest
layer are columnar. Their lateral surfaces are
bound to adjoining cells by desmosomes.
Meanwhile, their basal surfaces are attached
firmly to the basal lamina with the help of
hemidesmosomes and some anchoring proteins

ESTEBN\i & GOf\iZ/\LES' TEXTBOOK OF HISTOLOGY

and filaments. In between the epithelial cells Tunica Vasculosa (Uvea; Uveal
are numerous, free sensory nerve endings-the Tract)
reasons why the cornea is very sensitive.
The tunica vasculosa has three parts: 1)
The corneal epithelium has a very rapid
choroid, 2) ciliary body, and 3) iris'.
turnover rate of about 7 days. Consequently,
at any given time, many of its basal cells are The choroid refers to the vascular and
undergoing mitosis while many of its superficial pigmented layer that forms the middle histologic
cells are being shed. layer of the posterior %th of the eyeball. It
ranges in thickness from 0.1-0.2 mm. At its
Bowman's membrane refers to the thick
anterior edge, the choroid thickens and forms
(8-12 urn) fibrillar lamina to which the basal the Ciliarybody.
lamina of the corneal epithelium is anchored.
The iris, the third part of the tunica
This layer is acellular and amorphous. It consists
vasculosa, refers to the thin, circular diaphragm
of randomly-arranged type I collagen fibrils
that extends from the ciliary body. It surrounds
embedded in ground substance.
a round hole known as pupil.
The stroma (substantia propria), consisting
of dense regular connective tissue, comprises Choroid
90% of the cornea. It comprises of a ground
The outer surface of the choroids is
substance that is rich in keratan sulfate and
attached to the sclera by a thin layer of loose
chondroitin sulfate 'and where collagen fibers
connective tissue called suprachoroidal
(which are of uniform size) and cells-mostly lamina (epichoroid; lamina fusca). The
fibroblasts (called keratocytes) and a few suprachoroidal lamina is rich in elastic fibers.
lymphocytes-are embedded. The collagen Its cells are mostly fibroblasts and melanocytes
fibers are in bundles that are arranged in layers: but there is a sprinkling of macrophages which
Within each layer, the fibers are regularly are likewise present all over the uvea. The
arranged parallel to each other, but !he direction suprachoroidal lamina blends imperceptibly
of the fibers in each successive layer differs from with the substance of the choroid.
those in the layers that are below or above them.
The choroid has three distinct layers: 1)
The regular size and arrangement of its collagen
vessel layer, 2) choriocapillary layer, and 3)
fibers account for the transparency of the cornea Bruch's (glassy) membrane.
as opposed to the sclera.
Descemet's membrane is a homogenous
layer thinner (5-10 urn) than Bowman's
membrane. It is a basement membrane that
consists of the basal lamina of the endothelial
cells and some extracellular materials including
fine collagen fibers and a sprinkling of elastic
fibers.
The corneal endothelium is a simple low
cuboidal epithelium (squamous according to
many authors). The basal surface of its cells
rests on the basal lamina that, as stated in the
Fig. XIX-6. Choroid. The photomicrograph
preceding paragraph, forms part ofDescemet's
showsthe three histologic layersof the choroid:
membrane while their apical surface is in contact vessel layer, choriocapillary layer, and Bruch's
with aqueous humor. membrane.H&E x200.

EYE~
 The vessel layer is the outermost layer of the The pars plicata has numerous finger-like'
choroid. It consists of loose connective tissue projections called ciliary processes. Ciliary
where numerous branches and tributaries of the processes are made up of connective tissue that
ciliary arteries and veins are embedded. This is richly supplied with fenestrated capillaries
layer also contains many melanocytes. from which aqueous humor-the fluid that
The choriocapillary layer, the middle layer
fills the anterior and posterior chambers-
of the choroid, contains numerous fenestrated
is derived. The pars plana, on the other hand,
refersto the part ofthe epithelial portion devoid
capillaries that form a network. The capillaries
arise from the blood vessels that are present in
of ciliary processes.
the vessel layer. They are the largest capillaries The epithelial portion of the ciliary body
in the body. They supply the cells of the outer is covered posteriorly by the ciliary retina, a
layers of the retina with nutrients and oxygen. part of the anterior non-photosensitive portion
of the retina. The ciliary retina consists of two
Bruch's membrane (lamina basalis
epithelial celllayersthat are continuous with the
choroidea) is sandwiched by the choriocapillary
outer two cellular layers of the photosensitive
layer and the retina. It extends from the optic
portion of the retina (i.e., retina proper).
disc to the anterior edge (ora serrata) of the
photosensitive portion of the retina. The external layer of the ciliary retina
is the anterior continuation of the pigment
Bruch's membrane is 1-4 ~m thick. It is
epithelium of the retina proper. It consists of a
amorphous under the light microscope, but
singlelayer of simple cuboidal cells that contain
under the electron microscope, it has been
a lot of melanin. The internal layer, often
shown to consists of five layers. These layers,
referred to as ciliary epithelium, consists of
from the most external to the most internal, are
a simple cuboidal or columnar epithelium
as follows: basal lamina of the endothelium
whose cells do not possess melanin. The ciliary
of the capillaries of the choriocapillary layer:
epithelium is derived from the layer of rods and
a layer of collagen fibers, a layer of elastic
cones (see page 307) of the retina proper but,
fibers, another layer of collagen fibers: and
unlike the latter, it is not photosensitive.
basal lamina ofthe pigment epithelium of the
retina.

Ciliary Body
The ciliary body forms a ring of tissue on
the inner surface of the anterior portion of the
sclera. It extends from the scleral spur-=--an
annular structure which serves as the origin of
some of the ciliary muscle fibers and the anchor
of the trabecular network (meshwork)-to
the ora serrata of the retina. On meridional
section, it is shaped like a triangle whose base
faces the posterior chamber.
The ciliary body is divisible into two
regions: an epithelial portion that adjoins the
vitreous cavity and equator of the lens, and a Fig. XIX-7. Ciliary Body and Adjacent
uveal portion adjacent to the sclera. Structures. Note the ciliary processes which are
finger-like processes on the posterior surface of
The epithelial portion is further subdivided the ciliary body, and the zonule fibers that pass in
into the pars plana and the pars plicata. between the ciliary processes. H&E x40.

ESTEB!\N& CC)NZ;\LES'TEXTBOOK OF HISTOLOGY

 The cells of the ciliary epithelium that line
the free surfaces of the ciliary processes actively
transport certain constituents of plasma to the
posterior chamber and thus help form aqueous
humor.
The uveal portion of the ciliary body is
mainly made up of smooth muscle fibers that
comprise the ciliary muscles. The muscle
fibers are embedded in a loose connective tissue
stroma.
The muscle fibers of the ciliary muscle
mostly originate from the scleral spur. However,
they spread out in several directions. Some
muscle fibers are longitudinally-arranged. Their
function is to open up the trabecular network
(discussed further later in this chapter) to enable
the aqueous fluid to drain.
Some muscle fibers of the ciliary muscle,
on the other hand, are circularly-arranged.
These muscle fibers are responsible for
accommodation. When focusing for near
vision, they contract. In the process, the ciliary
body moves forward, the tension on the zonule
fibers (see page 310) that keep the lens in
place decreases, and the lens-because of the
elasticity of its capsule-increases 'Its curvature,
causing its focal length to shorten. Conversely,
in focusing for far vision, the ciliary muscle
relaxes, causing the lens curvature to decrease
and its focal length to lengthen. This enables the
eye to focus on distant objects.
The Ciliary muscle is supplied with
sympathetic and parasympathetic nerve fibers.
The parasympathetic nerve fibers stimulate the
ciliary muscle to contract, but the role of the
sympathetic nerve fibers is not very clear yet.
Iris
The iris is a highly-pigmented and vascular
diaphragm that is immediately anterior to the
lens. It covers the lens incompletely, leaving a
round central opening (pupil).
The iris regulates the amount of light that
enters the eye by adjusting the size of the pupil.
Fig. XIX-B. Iris and Lens. Note the eosinophilic
smooth muscle fibers near the free edge of the
iris which comprise the sphincter pupillae muscle.
H&E x100.
The substance of the iris is primarily made
up of loose connective tissue that is richly
supplied with capillaries and where numerous
melanocytes, fibroblasts, and smooth muscle
cells are embedded. The melanocytes in the
iris account for the color of the eye. If they are
few, the eye is blue, if they are numerous, the
eye is brown.
The anterior uneven surface of the iris
is covered by endothelial cells in the fetus
but these cells disappear in early childhood.
Thus, in older children and adults, the anterior
surface of the iris is covered, albeit incompletely,
not by endothelium, but by fibroblasts and
melanocytes.
The posterior surface of the iris, on the
other hand, is smooth and rests on the anterior
surface of the lens. It is lined by the iridial
retina, i.e., the anterior extension of the ciliary
retina. The internal layer of the iridial retina
(i.e., the layer that abuts on the lens), unlike
its corresponding layer in the ciliary retina, is
heavily pigmented. The cells of the external,
less pigmented layer of the ciliary retina,
meanwhile, are the myoepithelial cells.
They are arranged radially on the periphery
of the iris to' comprise the dilator pupillae
muscle which dilates the pupil when they are
stimulated by sympathetic impulses.
EYE~
 N ear the free margin of the iris, there are
smooth muscle fibers arranged in a circular
manner. These muscle fibers comprise the
sphincter pupillae muscle which constricts
the pupil when stimulated by parasympathetic
impulses,
Tunica Interna (Retina)
The retina, the innermost histologic layer of
the wall of the eyeball has two regions: anterior
retina and posterior retina (retina proper).
The anterior retina is not photosensitive. It
consists of the ciliary body and iridial retina
that have been described in connection with the
tunica vasculosa.
The retina proper, on the other hand, is the
photosensitive region of the retina. It extends
from the optic disc to the posterior edge of the
ciliary body where it ends-as a wavy line, the ora
serrata. The term retina, unless qualified, refers
to the retina proper.
Cells of the Retina
The cellular elements of the retina (proper)
consist of pigmented epithelial cells, neurons,
and supporting (glial) cells. ,.
There are several types of neurons in the
retina. Three of these neuronal types-rods
and cone cells, bipolar cells, and ganglion
ESTEBAN& CONZ;'\LES'TEXTBOOK OF HISTOLOGY
cells-synapse serially to span the whole'
breadth of the retina. Thus, the retina is only
three neurons thick. Also present in the retina
are two major types of association neurons: the
horizontal and amacrine neurons.
The supporting cells in the retina include
Muller cells, astrocytes, and microglia. The
Muller cells are large cell~ with numerous
processes that envelop the neurons of the retina.
The astrocytes are present only in the nerve
fiber layer while the microglia are found all
over the retina and serve as phagocytes.
Histologic Layers of the Retina
The cells of the (posterior) retina are well-
arranged and highly-organized. They form 10
histologically distinct layers. From the most
external to the most internal, these layers are
the following: 1) pigment epttheltum, 2)
layer of rods and cones: 3) outer limiting
membrane, 4) outer nuclear Iayer: 5) outer
plexiform layer: 6) inner nuclear layer: 7)
inner plexiform layer: 8) ganglion cell layer:
9). nerve fiber layer: and 10) inner limiting
membrane.
Pigment Epithelium
The pigment epithelium is made up of
a single layer of cuboidal cells that are richly
supplied with melanin. The cells rest on a basal
lamina which, as previously mentioned, forms
the innermost part of Bruch's membrane of the
choroid. The cellshave a basally-located nucleus.
Their basal plasma membrane has infoldings
with associated numerous mitochondria. Their
lateral surfaces are connected to those of the
adjacent cells by zonula occludens, zonula
adherens, desmosomes, and gap junctions. Their
~ical surfaces are provided with microvilli
Fig. XIX-9. The Neurons that Synapse Serially
to Span the Retina. The schematic diagram
illustrates that, in essence, the retina is only three
neurons thick because three types of neurons
synapse sequentially to span the layer. These
neurons are the rod and cone cells, bipolar cells,
and ganglion cells.
 inn&rlinllti~(I
IMfl'lbrane

lIangilOneoill
laYjlf

I(!net
pje)(JfOrrn IlIYet

Inner
nuclear ·llIyer

wlilr
pleXffilrml.~er

OlJtet
n.udMtlaye,
wiilrlimiill'!Q
membrano·

piQll\I!nl
OpIih"li!tm

Fig. XIX-10. Histologic Layers of the Retina. H&E x200.

and cylindrical processes, where the tips of the dendrites of the rods and cones cells comprise
photoreceptors that .comprise the next layer of the layer of rods and cones. Their nuclei
the retina fit into. constitute the outer nuclear layer of the retina
The pigment epithelium serves to increase while their axons synapse with the bipolar cells
the contrast of the visual image by absorbing in the outer plexiform layer.
light that would otherwise be reflected inward The rod cells are more numerous than
towards the photoreceptors. Its cells also the cone cells. Estimates place their number
phagocytose the ends or tips of the rods as they
are being renewed. They likewise synthesize
and store melanin and produce retinal, the
visually active form of vitamin A, from trans-
retinol.

Layer of Rods and Cones

This layer consists of rods and cones, the
light-sensitive dendrites of the rod and cone
cells, respectively. The more appropriate term
for the layer is photoreceptor layer which
describes its function.

Rod and Cone Cells

The rod and cone cells are specialized Fig. XIX-11. Rod and Cone Cells. These
bipolar neurons. They derive their name from photosensitive cells of the retina are
the shape oftheir photosensitive dendrites. The morphologically similar because their dendrites
consist of inner and outer segments. These
dendrites of rod cells are tubular in shape and
segments are connected together by a
are called rods. In contrast, those of the cone constricted region of the cytoplasm that is
cells taper apically and are called cones. The structurally akin to a cilium.

EYE~
at about 100-120 million per eye. They
are slender, elongated cells. Their rod has
two segments- inner and outer-whose
combined length is between 100-120 ~m.
The two segments are connected together by
a constricted region of cytoplasm that contains
nine microtubule doublets and whose structure
is similar to the cilium minus the central pair of
microtubules.
The inner segment has numerous
mitochondria and polyribosomes, and a well-
developed Golgi complex. The cylindrical
outer segment, on the other hand, is filled with
stacks of membrane-bound flattened disks. The
membrane of these disks contains the pigment
rhodopsin (visual purple). This is produced
when retinal (which, as previously pointed
out, is produced by the cells of the pigment
epithelium) combines with the protein opsin.
When hit by light, rhodopsin undergoes a
change in configuration. This event initiates the
visual stimulus. Immediately after it undergoes
configurational change in response to light,
rhodopsin is reconstituted. The rods are more
sensitive to light than the cones. They are also
the ones that the eye uses in conditions of poor
light (e.g., night-vision).
The flattened disks in the apex of the
rod cells are continuously shed off and
phagocytosed by the cells of the pigment
epithelium. To replace the disks being lost,
there is a continuous synthesis of disks in the
inner segment of the rods from where the disks
gradually migrate to the outer segment. It is
also in the inner segment where rhodopsin is
produced. Newly produced rhodopsin is
distributed evenly among all the flattened
disks in the outer segment of the rod.
The cone cells are also elongated cells,
but they are broader and shorter than the rod
cells.At 6 million per eye, they are considerably
fewer than the rod cells. Their basic structure
resembles that of the rod cells, except that their
dendrites (cones) taper apically.The cones,like
the rods, also consist of an outer segment and an
inner segment whose combined length is only
65 ~m to 75 ,~m.
ESTEBAN& CjOlNz/\LES'TEXTBOOK OF HISTOLOGY
Cones contain severalpigments responsive '
to green, red, and blue light. The principal
pigment that they possess is iodopsin which,
unlike rhodopsin (the pigment in rods), is
stimulated only by intense light. Iodopsin
is responsible for color perception and visual
acuity.
The flattened disks present in the outer
segment of cones, unlike those in rods, are not
shed off.
Outer Limiting Membrane
This is a narrow, eosinophilic region that
contains the junctional complexes that are
formed by the rod and cone cells with Muller
cells.
Outer Nuclear Layer
This is the area where the nuclei and cell
bodies of the rod and cone cells are lodged.
The nuclei of the cone cells are in the outer
region while those of the rod cells are in the
c~~tralregion of this layer.
Outer Plexiform Layer
This layer-amorphous under the
microscope-contains the axons of the rod and
cone cells, the dendrites of the bipolar cells,
and the synapses they form. The axons of cone
cells synapse one-on-one with the dendrites of
bipolar cellswhile the axons of severalrods may
synapse with the dendrite of a single bipolar
cell.
Horizontal neurons also synapse with the
axons of the rod and cone cellsin this layer.
Inner Nuclear Layer
This region is slightly thinner than the
outer nuclear layer. It contains the cell bodies
and nuclei of the bipolar cells. In addition, it
also includes the cell bodies of the horizontal
and amacrine neurons, and the nuclei of the
Muller cells, whose processes extend from
the outer limiting to the inner limiting
membranes.
Inner Plexiform Layer
This region contains the synapses formed
by the axons of the bipolar cells with the
highly-branched dendrites of the ganglion
cells. Likewise, the ganglion cells synapse with
the amacrine neurons in this layer.
Ganglion Cell Layer
The bodies and nuclei of the ganglion
cells comprise this layer. Also present in this
layer are the retinal vessels.
Nerve Fiber Layer
This region contains the unmyelinated
axons (afferent fibers) of the ganglion cells.
The axons gather in bundles that run parallel to
the retina to converge in the optic disc and form
the optic nerve.
Inner Limiting Membrane
This is the layer of the retina adjoining
the vitreous body. It consists of the basement
membrane of the Muller cells.
Modifications of the Retina.
Some areas in the retina do not exhibit the
10 histologic layers that typify this innermost
coat of the eye. For example, the anterior retina
(i.e., ciliary and iridial retina), as previously
described, has only two layers.
Another atypical retinal area is the fovea
(fovea centralis). It is an ovoid depression
that is about 0.5 mm in diameter. It is at the
same level as the bottom half of, but about 4
mm lateral to, the optic disc. The fovea is the
area of the eye where visual acuity is greatest
and color perception is finest. It is packed with
photoreceptors exclusively made up of cones.
In the fovea, the light rays fall directly on the
photoreceptors because the inner layers of the
retina are absent.
Surrounding the fovea is an ovoid area that
has a diameter of 1.5 mm. It is called the macula
lutea (yellowspot) and isyellow in color in gross
specimens. The color is imparted by carote~oids
present in the area. Carotenoids also serve to
absorb the excess blue and ultraviolet light that
enters the eye.
Still another atypical area in the retina is
the optic disc. The optic disc is the area where
the axons of the ganglron cells of the retina
(i.e., optic nerve fibers) converge and exit the
eye. The optic disc, about 3 to 4 mm to the nasal
side of the fovea, is about 3 mm in diameter. It is
referred to as the blind spot of the eye because
it has no photoreceptors.
Blood Supply of the Retina
The central artery of the retina, a branch of
the ophthalmic artery, supplies the inner layers
of the retina. The outer layers of the retina, as
has been previously mentioned, are supplied by
the capillaries of the choriocapillary layer of
the choroid. This layer consists of branches of
the short ciliary and posterior ciliary arteries
that also come from the ophthalmic artery.
The capillaries of the retina drain into the
retinal vein, a short vein that empties into either
the superior ophthalmic vein or cavernous
sinus.
The central artery of the retina enters, and
the retinal vein exits, the eye through the center
of the optic nerve.
REFRACTIVE MEDIA AND
CHAMBERSOF THI EYIE
Refractive media of the eye refers to the
components of the organ that serve as a system
of lenses that focuses an inverted image of the
object being viewed on the photoreceptors in
the retina. From the most anterior to the most
posterior, the refractive media consists of the:
1) cornea, 2) aqueous humor, 3) lens, and 4)
vitreous humor (vitreous body).
Lens
The lens is a biconvex elastic structure that
is more convex on its posterior than its anterior
EYE~
 subcapsular
epithelium
Fig. XIX-12. Lens. The lens is enveloped by a lens
capsule. Deep in the capsule is a simple cuboidal
epithelium, calle-d subcapsular epithelium,
present on the anterior surface of the lens
only. The cortex is formed by cells called lens
fibers that are devoid of nuclei and cytoplasmic
organelles.
surface. It is highly cellular but transparent and
amorphous. It is enveloped by a 10-20 ~m thick
capsule (lens capsule) made up of collagen
fibers (type IV) and glycoproteins.
Deep in the lens capsule) the anterior
surface of the lens is covered by a simple
cuboidal epithelium (subcapsular epithelium).
The cells of this epithelium are bound together
by desmosomes and gap junctions. They
are nucleated and their cytoplasm has few
organelles. The subcapsular epithelium covers
only the anterior surface of the lens. It is lacking
on the posterior surface.
The substance (cortex) of the lens consists
of 2)000 to 3)000 specialized cells called lens
fibers. The lens fibers have no nuclei and
cytoplasmic organelles. Their cytoplasm is
filled with a protein called crystallin.
There is hardly any intercellular substance
in between the lens fibers because the cell
membranes of adjacent lens fibers are fused.
The lens fibers are very long (7-10 mm) and
they span the anterior and posterior poles of the
lens. They are 8-10 ~m in width and 2 ~m in
thickness. They come in the shape of six-sided
prisms with the more peripheral fibers curving
to give the lens its biconvex form.
ESTEBAN& GONZALES' TEXTBOOK OF HISTOLOGY
The lens fibers differentiate from the cells'
of the subcapsular epithelium. Their production
is continuous throughout life) although at a
much slower rate as the person ages.
The lens lies posterior to the iris. It is kept in
place by zonule fibers (suspensory ligament).
The zonule fibers are fibrillar structures made
up of bundles of microfibrils (fibrtllm) that are
identical to those prese-nt in elastic fibers. They
originate from the basement membrane of the
ciliary epithelium) course in between the ciliary
processes on their way to the lens) and insert
into the lens capsule.
Chambers of the Eye and the
Aqueous and Vitreous Humors
The cavity of the eyeball is divided into
three compartments (chambers; cavities)
that are filled with transparent material. The
compartments include 1) anterior chamber) 2)
posterior chamber) and 3) vitreous chamber.
The anterior chamber refers to the space
bounded by the cornea anteriorly and the iris
~~d lens posteriorly. The posterior chamber)
-I
on the other hand) refers to the space bounded
by the iris anteriorly and the Ciliary processes
and zonule fibers laterally and posteriorly. The
vitreous chamber) meanwhile) refers to the large)
spherical compartment that is behind the lens)
zonule fibers) and ciliary processes.
The anterior and posterior chambers are
filled with aqueous humor while the vitreous
chamber is filled with vitreous humor.
Aqueous humor is a clear) slightly alkaline
fluid derived from blood plasma within the
capillary network of the ciliary processes.
Compared with plasma) it contains less
protein) urea) and glucose) but it has more
lactate) pyruvate) and ascorbate. Its electrolyte
composition is also different from that of
plasma. The anterior chamber contains about
250microliters and the posterior chamber about
60 microliters of aqueous humor.
Aqueous humor is secreted continuously
into the posterior chamber by the ciliary
Fig. XIX-13. Eyelids and Conjunctiva. The
eyelids are thin folds of tissues that cover the
exposed anterior surface of the eyeball. The
palpebral
conjunctiva, on the other hand, refers to the conjunctiva
mucous membrane that covers the exposed
region of the sclera (bulbar conjunctiva) and bulbar
lines the inner surface of the eyelids (palpebral conjunctIva
conjunctiva). The bulbar and palpebral
conjunctivae are continuous with each other at
the superior and inferior fornices.

epithelium. From the posterior cliamber,

aqueous humor flows into the anterior chamber
via the pupil and drains through the trabecular
network (trabecular meshwork) and the canal
of Schlemm to the veins at the limbus. Aqueous
humor has a turnover rate of I.S hours. Aside
from acting as a refractive medium, it also serves
to nourish the cornea.
The vitreous humor is the gelatinous but
transparent mass that fills the vitreous cavity.
It touches against the aqueous humor in the
posterior chamber but the two do not mix.
The vitreous humor is 99% water. It
contains collagen fibrils (mostly type II and
type XI) and glycosaminoglycans (principally
hyaluronic acid) that are heavily hydrated. A
few cells (hyalocytes)which may be responsible
for synthesis of collagen and hyaluronic acid
are also present in the peripheral region of the
vitreous humor.
Aside from serving as a refractive medium,
the vitreous humor serves as a pathway for
nutrients to the lens, ciliary body, and retina. It
also provides structural integrity to the eye-it
pushes against the retina and helps hold it in
place.
Vitreous humor is not continually
replenished like aqueous humor. The loss of
vitreous humor (as when the eye is lacerated) or
shrinkage (as what sometimes happens among
the elderly) can result in retinal detachment
(i.e., the retina detaches from the choroid:
actually the separation usually occurs between
the pigment epithelium and the rods and cones
layer). Retinal detachment invariably leads to
blindness unless immediately treated.
ACCESSO TRUCTUR 5
(ADNEXA) OF THE EYE
The accessory structures of the eye
include the extra-ocular muscles (EOMs),
conjunctiva, ,and lacrtmal apparatus. The
extraocular muscles are skeletal muscles that
have been mentioned in passing at the start of
this chapter. Their attachments and functions
are discussed in gross anatomy textbooks.
Conjunctiva
Conjunctiva refers to the thin, translucent
mucous membrane that covers the exposed
anterior portion of the sclera and lines the inner
surface of the eyelids.
The conjunctiva that overlies the sclera is
referred to as bulbar conjunctiva while the
one that lines the eyelids is termed palpebral
conjunctiva. In the transitional areas (superior
and inferior fornices) between the bulbar and
palpebral conjunctivae, the conjunctiva adheres
loosely to the underlying tissue. The space
bounded by the palpebral conjunctiva anteriorly
and by the bulbar conjunctiva posteriorly is
known as the conjunctival sac. This sac receives
ocular medications (e.g., eye drops).
Histologically, the conjunctiva consists
of an epithelium that is stratified columnar

EYE CUI
Fig. XIX-14. Conjunctiva.
The photomicrograph
shows the transitional
area (i.e., fornix) between
the palpebral and bulbar
conjunctivae. The epithelium
of the conjunctivae is
stratified columnar that has
numerous goblet cells. H&E
x40. .

but atypical because it has numerous goblet

cells, and a lamina propria made up of loose
connective tissue.

muscle
tissue
Fig. XIX-1S. Eyelid. The eyelid is covered
externally by skin and lined internally by
mucous membrane (i.e., bulbar conjunctiva). In
the photomicrograph, the dermis shows a hair
follicle and its associated sebaceous glands, also
referred to as glands of Zeis. Near the mucosal
surface are atypical sebaceous glands called
meibomian glands that are embedded in the
tarsal plate, a dense fibroelastic structure that
forms the core of the eyelid. The muscle fibers
comprise the orbicularis oculi muscle, and in the
upper lid, the levator palperbra. H&E x100.
.Eyelids (Palpebrae)
The eyelids (upper and lower) are the thin
folds of tissues that protect the anterior exposed
portion of the eyeball. The external surface of
each eyelid is covered by skin while its internal
surface is lined by palpebral conjunctiva. The
skin contains some fine hairs, except at the free
edge of the eyelid where 2-3 rows of coarse
long hairs (eyelashes) project anteriorly. The
hypodermis of the eyelids is formed by very
loose connective tissue that is devoid of fat.
The core of the eyelid consists of a dense
fibroelastic plate (tarsus, tarsal plate). Anterior
to the tarsal plates, and separated from them by
small amount of connective tissue, are skeletal
muscle fibers that comprise the orbicularis
oculi muscle. In the upper lid, the skeletal
muscle fibers form a second muscle, the levator
palpebrae.
There are several types of glands present
in the eyelids, the more important ones are the
meibomian, Moll, and Zeis.

ESTEBAN & (1CNZf\LES' TEXTBOOK OF HISTOLOGY

 The meibomian glands-numbering
20-25 per eyelid-are sebaceous glands that
are atypical because they are not associated
with hair follicles. They also have very long
ducts into which numerous acini drain. They
are located within the tarsal plate and their
ducts open at the free edge of the eyelids. The
tarsal plate also houses some sweat glands
and the Ciaccio's glands (Wolfring's glands)
which are accessory lacrimal glands whose
secretions are delivered to the conjunctival
surface.
The glands of Zeis are sebaceous glands
that are smaller than the meibomian glands
while the glands of Moll are modified apocrine
sweat glands. The glands of Zeis and Moll are
located in the dermis and their ducts empty into
the follicles of the eyelashes. Their secretions
and those of the meibomian glands combine to
form the oil layer that makes up the third layer
of the tear film that protects the cornea.
Incidentally, the tear film that lubricates
the eyelid and prevents the cornea from drying-
up consists of three layers: mucin layer, water' , one set per eye, involved in the production
or aqueous layer, and oil layer (see preceding
paragraph).
The mucin layer (precorneal tear
film), as previously described in connection
with the cornea, soaks the microvilli of
the most superficial layer of cells of the
corneal epithelium. It is made up of lipid and
glycoprotein that aids in ensuring th~t the tear
film adheres to the eye.
The water or aqueous layer forms the middle
layer of the tear film. It is what is usually referred
to as "tears." It is a fluid similar in composition
to blood plasma and is produced by the lacrimal
glands. It forms a film that moistens the exposed
surface of the eye. It also nourishes the corneal
epithelium and mechanically removes irritants
and other foreign substances from the corneal
surface. It likewise prevents corneal infection
because it contains antibacterial substances,
lymphocytes, and other white blood cells.
The oil layer helps prevent evaporation of
the aqueous layer.
Lacrimal Apparatus
Lacrimal apparatus refers to the structures,
and drainage of tears (i.e., the aqueous layer
of the tear film). It consists of the lacrimal
gland, lacrimal canaliculi, lacrimal sac, and
nasolacrimal duct.

Ilcrlmat
clnaHcul···

;8.
lacrl

Fig. XIX-16. Lacrimal Apparatus. The lacrimal apparatus consists of the lacrimal gland which
produces tears, and the system of tubes that drain tears into the nasal cavity (i.e., lacrimal canaliculi,
lacrimal sac, and lacrimal duct).

EYE"

-------- --------- -------------------------------------


Lacrimal Gland
The lacrimal gland is a serous gland
located in the anterosuperior temporal portion
of the orbital cavity. It consists of several lobes
that empty their secretion into the snperior
conjunctival fornix via 6-12 excretory ducts.
The lacrimal gland is a tubuloalveolar
gland structurally similar to the parotid gland.
Its secretory portions consist of typical serous-
secreting cells that rest on a basal lamina.
As in many other exocrine glands, there are '. when the lacrimalgland produces a lot of tears
myoepithelial cells associated with the
secretory portions of the gland. .,.:J
Lacrimal Canaliculi, Lacrimal Sac,
and Nasolacrimal Duct
Tears that are continuously produced
by the lacrimal gland are spread over the eye
by the eyelids when the person blinks. They
drain into the lacrimal canaliculi, two tiny
tubes (superior canaliculus and inferior
& GONZALES' TEXTBOOK
ESTEB,L".J~ OF HISTOLOGY
Fig. XIX-17. Lacrimal Gland. The
lacrimal gland is divided into lobules
by connective tissue septa (5). The
lobules are occupied by the serous
acini (a) that produce tears. The acini
are drained by a system of ducts
which include interlobular ducts
(ild) embedded in the connective
tissue septa. The septa are also
used as passageways by the blood
vessels that supply the organ, such
as the interlobular artery (ila) in the
photomicrograph. H&E x100.
canaliculus) that are about 1 mm in diameter
and 8 mm long. The round openings (lacrimal
puncta) of the lacrimal canaliculi are located
on the medial aspect of the upper and lower
lid margins. The canaliculi are lined by thick
nonkeratinized stratified squamous epithelium.
They terminate by merging to form a common
canaliculus that drain into the lacrimal sac, a
dilated portion of the lacrimal drainage system
that lies in the lacrimal fossa of the frontal
bone. The lacrimal canaliculi are so tiny that
(e.g., as when a person cries), the tears cannot
get accommodated by the lacrimal canaliculi
and some find their way into the person's face.
The inferior continuation of the lacrimal
sac is the- nasolacrimal duct which opens
into the inferior nasal meatus lateral to the
inferior turbinate. The connection between
the lacrimal drainage system and the inferior
nasal meatus is the reason why crying results in
a runny nose.
E r

T
he ears are a pair of complex organs
that contain the receptors for two middle
,ear
important sensory functions: liearing
and equi i6ration (i.e.,-sense of balance or
equilibrium). .

All three regions of the ear are involve a

witli tlie sense of nearing. The external ear inner
eardrum ear
eollects sound wave and directs' them into
the middle ear. The mioole ear then amplifies Fig. XX-1. Ear. Schematic diagram shows the
the vibrations that the sound waves create and three regions of the ear: external ear which
transmits them to the inner ear. Meanwhile, consists of the auricle and external auditory
receEtors in tne inner ear convert the sound meatus; middle ear which is separated from the
external ear by the eardrum and contains the
rviorations into nerve imEulses that are then
auditory ossicles; and inner ear which includes
transmitteJ via tile codilear aivision of tne the receptors for the senses of hearing and
;vesti1>uloclJchlear nerv (eN VIII; auditory equilibrioception. The middle ear is connected
nerve; acoustic nerve) to tne auditory:center to the nasopharynx by the auditory tube.
of the brain for interpretation.
EXTERNAL EAR
Eg_uilioration on the other hand, helps
prevent one from falling over when one is erect
or walking. It involveson1x:tne inner ear,where
the receptors for this physiological sense are
located. The nerve impulses generated by the
receptors for the sense of balance in the inner
ear are transmitted to the vestibular center, of
the brain for interpretation by:tIie vestitiula~
<livision of the vestihuJocochlear nerve (eN
VIII).

Fig. XX-2. Middle Ear. Note that the auditory
ossicles articulate with each other to form a chain
that spans the middle ear. Note further that the
medial wall of the middle ear-which separates
it from the inner ear-has two openings, round
window and oval window, covered by membrane.
The framework of the auricle is formed
by a single piece of elastic cartilage (auricular
cartilage), exceptin its inferior part, the earlobe,
where the framework is connective tissue.
The auricular cartilage is attached to the skull
and surrounding structures by ligaments and
rudimentary skeletal muscles. It is covered by
skin that possesses fine hair, sebaceous glands,
and a fewsweat glands. The skin adheres closely
to the perichondrium of the auricular cartilage
anteriorly, but posteriorly, some amount of
subcutaneous tissue existsbetween the skin and
the perichondrium.
The framework of the outer third of the
wall of the external auditory meatus is elastic
cartilage that is continuous with the auricular
cartilage while the framework of the inner two-
thirds is bone.
The cartilage and bone that frame the
external auditory meatus are lined by skin
that is reflected into the external surface of the
tympanic membrane. In the cartilaginous part
of the canal, the skin contains coarse hair, large
sebaceous glands, and modified sweat glands
called ceruminous glands that extend into the
ESTEB/\\l & C30\iZ/\LES' TEXTBOOK OF HISTOLOGY
subcutaneous tissue. The ceruminous glands'
produce a waxy secretion called cerumen and
their ducts open into the skin surface or into
ducts of the sebaceous glands. In the osseous
part of the canal, the skin is thin and lacks hair
follicles and sebaceous glands, and adheres
closely to the periosteum of the underlying
bone. .
MIDDLE EAR (TYMPANIC
CAVITY)
The middle ear consist ofanarrow,irregular,
air-filled chamber in the temporal bone, and the
three tiny bones (auditory ossicles) and two
small skeletal muscles that are lodged therein.
The bony framework of the middle ear is lined
by mucosa that, for the most part, consists of a
ciliated simple cuboidal epithelium with goblet
cells and an underlying thin lamina propria.
The middle ear is separated from the
external acoustic meatus by the eardrum
(tympanic membrane) and connected to the
nasopharynx by the auditory tube. Posteriorly,
it communicates with the mastoid antrum,
an air sinus in the petrous temporal bone that
leads into the mastoid air sinuses. The mastoid
antrum and air sinuses are lined by mucosa that
is continuous and similar to the mucosa that
lines the middle ear.
The medial wall of the middle ear contains
a rounded bulge or prominence (promontory)
and two openings that are closedby membrane:
the oval window (vestibular window; fenestra
vestibuli; fenestra of the vestibule, fenestra
ovalis) and the round window (cochlear
window; fenestra cochlea; fenestra of the
cochlea; fenestra rotunda). The oval window
is posterosuperior to the promontory while the
round window is posteroinferior to it.
Eardrum (Tympanum; Tympanic
membrane)
The eardrum is a thin fibrous membrane
attached to the surrounding bone by a
fibrocartilaginous ring. It has three histologic
layers.
The middle layer is a dense connective
tissue mainly made up of collagen fibers that
are embedded in a small amount of extracellular
material and arranged in two layers:outer radial
and inner circular.
The outer layer is thin skin that is
continuous with the skin that lines the external
auditory meatus. This skin has a very thin
dermis devoid of hair folliclesand glands.
The inner layeris mucosa that is continuous
with the mucosa that lines the middle ear. In
the eardrum.' however, the simple cuboidal
epithelium is devoid of cilia and goblet cells.
Auditory Tube (Eustachian tube;
Pharynqotympanic tube)
The framework of the auditory tube is
formed by bone in the segment of the tube near
th e nuiddl e ear an db y car til1 age m
. th e segment
near th e nasoph arynx. It IS' lime db y mucosa th at
.IS contimuous WIith th at 0 fth e nasoph arynx and
the middle ear. The epithelium of the auditory
tube is pseudo stratified ciliated columnar to
much of its length with goblet cells from its
opening in the nasopharynx. It becomes simple
cuboidal with a few goblet cells near the middle
ear. The lamina propria contains mixed glands
and is richly supplied with MALT, including'
lymphoid nodules, especially in the segment of
the tube near the nasopharynx where they are
referred to as tubal tonsil.
The auditory tube which becomes patent
with swallowing helps equalize pressure on
both sides of the eardrum by allowing outside
air to enter the middle ear.
Auditory Ossicles
The three auditory ossiclesare attached to
the wall of the middle ear by ligaments. They
are invested by mucosa similar to that of the
eardrum. They are named according to their
shape: hammer (malleus), anvil (incus), and
-- -_- --- --------
stirrup (stapes). The hammer is the largest of
the ossicles.It consists ofa head, neck, anterior
and lateral processes, and a handle. The anvil
consists of a body and long and short limbs.
The stirrup consists of a head, anterior and
posterior limbs, and a footplate or base.
By articulating with each other via freely
movable joints, the ossicles form a bony chain
that spans the lateromediallength ofthe middle
ear. Their articulating surfaces are lined by
hyaline cartilage.
The handle of the hammer is attached to
the eardrum while its head articulates with the
body of the anvil. The long limb of the anvil
articulates with the head of the sti~rup, whose
footplate fits into the oval window where it is
secured by the annular ligament. The oval
window is an orifice that connects the middle
ear to the vestibule of the inner ear, but is
completely and permanently closed by the
footplate of the stirrup and its annular ligament.
The ossicles mechanically amplify and
transmit sound wavesfrom the external surface
of the eardrum to the inner ear via the oval
window.
Skeletal Muscles in the Middle
Ear
The two tiny skeletal muscles in the
middle ear-likewise invested by a thin
mucosa that is similar to that which envelops
the ossicles-are the tensor tympani and
stapedius. The tensor tympani is attached
to the malleus while the stapedius is attached
to the stirrup. The two muscles contract in
response to loud or sudden sounds to prevent
excessive movement of the ossicles, which
could damage the delicate structures in the
middle ear.
I E EAR
The inner ear consists of a set offluid-filled
bony cavities within the petrous part of the
temporal bone, collectively referred to as the
bony labyrinth, and the likewise fluid-filled
EAR .,.
--------------~---~
 roembranoqs labynf)ij1
Fig. XX-3. Inner Ear. The inner ear is made
up of the bony labyrinth and the membranous
labyrinth. The bony labyrinth refers to the
cavities in the petrous temporal bone filled
with fluid called perilymph. Meanwhile, the
membranous labyrinth refers to the fluid-filled
and membrane-bound structures found in
the bony labyrinth and bathed by perilymph.
The bony labyrinth has three parts: vestibule,
three semicircular canals, and cochlea. The
membranous labyrinth, on the other hand,
has two components: cochlear duct and
vestibular apparatus (consisting of two sacs,
the saccule and utricle, and three semicircular
ducts). The fluid that fills the membranous
labyrinth is called endolymph.

membrane-bound structures that they contain, Vestibule

collectively referred to as the membranous
labyrinth.
The fluid that fills the bony labyrinth is
called perilymph. It is similar to CSF and is
widely believed to be an ultrafiltrate of plasma,
but the place where it originates and is absorbed
has not yet been established.
The fluid that fills the cavity of the
membranous labyrinth, on the other hand,
is called endolymph. Endolymph differs
markedly in composition from perilymph. It has
a high K+content and is similar to intracellular
fluid in composition.
Bony labyrinth
The bony labyrinth is located between the
middle ear laterally and the internal acoustic
meatus medially. The internal acoustic meatus
is a canal that leads to the cranial cavity and
serves as a passageway for, among other
structures, the facial nerve (CN VII) and the
vestibulocochlear nerve (CN VIII).
The bones that frame the bony labyrinth
are provided with periosteum that is overlaid
by mesothelium.
The bony labyrinth has three parts:
vestibule, semicircular canals, and cochlea.
The vestibule forms the center of the bony
labyrinth.
Its lateral wall which is also the medial wall
of the middle ear contains two orifices: the
ovalwindow and the round window. The oval
window, as previously mentioned, is closed by
the footplate of the stirrup and its anchoring
ligament, the annular ligament, while the round
window is closed by a fibrous membrane, the
secondary tympanic membrane.
The medial wall of the vestibule, on the
other hand, contains an orifice that leads into
.the vestibular aqueduct, a tiny canal that
opens into the posterior surface of the petrous
part of the temporal bone. It also contains the
endolymphatic duct, a part of the membranous
labyrinth.
Semicircular Canals
There are three semicircular canals:
anterior, posterior, and lateral. Each is about
1 mm in diameter and forms an incomplete
circular tube that has two ends that project
posterosuperiorly from the vestibule. One of
the two ends of each semicircular canal exhibits
a dilatation called ampulla, whose diameter is
about twice that of the rest of the tube. The
semicircular canals are oriented vertically

ESTEB;\\j & CJC)f~ZN_ES' TEXTBOOK OF HISTOLOGY

and are positioned such that each canal is at
an angle of about 100 degrees relative to the
others.
I
Cochlea
The cochlea is a spiral tunnel that is 30-
34 mm long. It coils 21/2 to 23,4 turns around a
piece of bone (modiolus) to form a structure
that looks like a snail's shell that has a wide
base and a tapered apex. Its base is about 9 mm
in diameter while its height (i.e.) base-to-apex
distance) is about 5 mm. The base ofthe cochlea)
oriented posterornedially, is continuous with
the vestibule. The apex) oriented anterolaterally,
on the other hand) ends blindly.
Fig. XX-4. Cochlea. This scanning
photomicrograph shows a structure that looks
like a snail's shell with several large cavities. The
cavities comprise several turns of the cochlea
which is a spiral tunnel that coils 2% to 2%
turns around a piece of bone (modiolus). Each
turn of the cochlea has three compartments:
scala vestibule (sv), scala media (sm), and scala
tympani (st). The scala media is the cochlear
duct and, in life, is filled with endolymph. The
scala vestibuli and scala tympani, on the other
hand, are filled with perilymph. Note the cochlear
nerve (cn) that occupies the central area of the
photomicrograph. H&E x40.
The modiolus that serves as a pillar around
which the cochlea spirals consists of spongy
bone. It is conical and constructed like a screw
that has a spiral thread projecting from its lateral
surface. This spiral thread) called osseous
spiral lamina) projects about halfway into the
cavity of the cochlea. .
The modiolus contains the spiral ganglion
made up of the cell bodies of bipolar sensory
neurons whose dendrites (also called sensory
axons because they are axons structurally
but dendrites functionally) are in the organ
of Corti; and the efferent fibers (axons) of
inhibitory neurons whose terminations are also
in the organ of Corti.
The axons of the bipolar neurons together
with the efferent fibers (axons) of the inhibitory
neurons comprise the cochlear nerve. This
nerve joins the vestibular nerve in the internal
acoustic meatus to form the vestibulocochlear
nerve (auditory nerve; CN VIII) that emerges
from the meatus to enter the cranium.
Along the outer wall of the cochlea) the
periosteum is thickened to form the spiral
ligament which extends incompletely to the
osseous spiral lamina. The wide gap that
separates the osseous spiral lamina from
the spiral ligament is bridged by a thin sheet
offibrous connective tissue) the basilar
membrane.
The cochlear duct (i.e., the part of the
membranous labyrinth within the cochlea)
occupies a small region along the outer wall
of the cochlea. Its inferior wall rests partly
on the spiral limbus (i.e., the thickened
periosteum that lines the superior surface of
the osseous spiral lamina) but mainly on the
basilar membrane. There are) therefore) three
longitudinal channels within the cochlea: the
endolymph-filled cavity of the cochlear duct
that is otherwise known as the scala media, the
perilymph-filled scala vestibuli that lies above
the scala media; and the scala tympani that
lies inferior to the scala media and is filled with
perilymph.
EAR_
 The scala vestibuli and scala tympani
communicate with each other at the apex
of the cochlea through a small opening, the
helicotrema. At the base of the cochlea,
the scalae vestibuli and tympani continue
into the vestibule where the scala vestibuli
communicates with the oval window and the
scala tympani with the round window.
Thus, the scala vestibuli and scala tympani
comprise a single, albeit coiled, tube that begins
at the oval window and ends at the round
window. Its halfway point is marked by the
helicotrema.
Also in the cochlea, near the round window,
is the opening of the cochlear aqueduct
(cochlear canaliculus), a small can~l that also
contains perilymph and connects the scala
Fig. XX-So Compartments of the Cochlea. This
low-power photomicrograph shows the three
compartments of the cochlea: scala vestibuli,
scala media, and scala tympani. The scala
vestibuli is separated from the scala media by the
vestibular membrane (vm) while the scala media
is largely separated from the scala tympani by
the basilar membrane (bm), where the organ
of Corti-which contains the auditory receptor
cells-rests. The other labeled structures in the
photomicrograph are the stria vascularis (sv),
spiral ligament (slig), osseous spiral lamina (slam),
afferent nerve fibers (n) that are on the way from
the organ of Corti to the spiral ganglion, and
spiral limbus (sl). H&E x100.
ESTE::3f.\N& cC)r~Z!\LES' TEXTBOOK OF HISTOLOGY
tympani with the subarachnoid space in the
posterior cranial fossa. A dural sheath extends
into the cranial end of the aqueduct for a varying
distance while the rest of the aqueduct contains
loose connective tissue and mayor may not have
a lumen.
Membranous Labyrinth
The membranous labyrinth consists of
interconnected membrane-bound circular
tubes (ducts) and flattened tubes (sacs) that
are suspended in the perilymph. It has the
same general form as the bony labyrinth, and
in certain areas, is fixed to the wall of the latter.
The membranous labyrinth has two
components: 1) the cochlear duct which
contains the sensory structures for hearing; and
2) the vestibular apparatus which contains the
sensory structures for balance or equilibrium.
The vestibular apparatus is made up of two
sacs, the utricle and the saccule, and three
semicircular ducts (anterior, posterior, and
lateral).
The cochlear duct, as previously
mentioned, is within the cochlea of the bony
labyrinth, although part of it extends into the
vestibule. The semicircular ducts are within
their corresponding semicircular canals while
the utricle and saccule are in the vestibule.
The wall of the membranous labyrinth
consists of an epithelium supported externally
by connective tissue that varies in amount
depending on its location. In jhe cochlear duct,
there is minimal connective tissue to none.
Cochlear Duct
The cochlear duct is triangular in cross
section. Its upper end is blind and is attached
to the apex of the cochlea. Its lower end extends
into the vestibule and is connected to the lower
end of the saccule by a tiny tube, the ductus
reuniens (canaliculus reuniens, canalis
reuniens, Heusen's canal, Heusen's duct,
uniting canal).
 The outer wall of the cochlear duct, called
stria vascularis, consists of a stratified cuboidal
epithelium. This epithelium is atypical because
it contains capillaries among its cells. The stria
vascularis adheres to the highly vascular upper
part of the spiral ligament. It produces the
endolymph contained in the scala media.
The roof of the cochlear duct which
consists of a single layer of squamous cells
(mesothelium) adheres to the mesothelium
that lines the luminal surface of the scala
vestibuli. Together, the two mesothelial
layers comprise the vestibular membrane
(Reissner'smembrane) that separates the scala
vestibuli from the scala media (i.e., cavity of the
cochlear duct).
The floor or inferior wall of the cochlear
duct is related to the scala tympani and, as
mentioned earlier, rests mainly on the basilar
membrane and partly on the spiral limbus.
Incidentally, the basilar membrane and the
osseous spiral lamina are lined on their
undersurface by the mesothelium that coats
the luminal surface of the scala tympani. '.
The portion of the floor of the cochlear
duct that lies on the basilar membrane consists
of a highly specialized epithelium, the organ
of Corti. Meanwhile, the portion on the
spiral limbus consists of a simple squamous
or cuboidal epithelium, some of whose cells
(interdental cells) produce the components of
the tectorial membrane.
Organ of Corti
The organ of Corti is made up of sensory
cells (hair cells) and supporting cells. The
supporting cells are of severaltypes and include
pillar cells (rod cells) and phalangeal cells.
The central area of the organ of Corti is
occupied by a triangular canal, the inner tunnel
(tunnel of Corti), bounded on each side by a
single row of pillar cells. The pillar cells along
the inner wall of the tunnel are called inner
pillar cells while those that are along the outer
wall are called outer pillar cells. The inner
pillar cells number about 6,000 while the outer
pillar cells number about 4,000.
Pillar cells are tall columnar cells that have
expanded bases resting on the basal lamina.
The inner and outer pillar cells sla~t towards
each other and their apices come into contact
to enclose, together with the basilar membrane,
the tunnel of Corti. The nucleus of the pillar
cells is basally located. Electron micrographs
show that pillar cells contain a collection of
microtubules in their cytoplasm that look like
pillars, thus the name.
The hair cells in the organ of Corti are tall
columnar cells. They are about 16,000 to 20,000
and have minimal capacity to regenerate. About
a quarter of them, the inner hair cells, form a
single row internal to the inner pillar cells while
the rest, the outer hair cells, form three to five
rows external to the outer pillar cells. Like the
pillar cells, the inner and outer hair cells incline
towards each other.
The hair cells are called as such because
their apical surface is provided with stereocilia.
Their basal surface, on the other hand, form
synapses with the dendrites (sensoryaxons)
of the bipolar sensory neurons whose cell
bodies are in the spiral ganglia and the axon
terminations of inhibitory neurons from the
brain stem. Typically, a sensory axon synapses
with several hair cells and conversely, each hair
cell synapses with the sensory axons of several
neurons. The sensory axons of the bipolar
neurons and the axons of the inhibitory neurons
also synapse with each other and form a plexus
of nerves in the area.
The inner hair cells possess 50-60
stereocilia which, in electron micrographs are
seen to be arranged in three rows of ascending
height from the inner to the outer aspect of the
cell. They are surrounded by inner phalangeal
cells which also separate them from the basal
lamina.
The outer hair cells are about twice as
tall as the inner hair cells. Like the inner hair
cells, they are also surrounded by phalangeal
EAR"
 Fig. XX-6. Rec;eptor Organs of the Vestibular
Apparatus. The cristae ampullaris and
maculae contain the receptors for the sense
of balance. There are three cristae ampullaris.
They are located in each of the ampullae of
the three semicircular ducts. There are two
macula, macula of the saccule and macula
of the utricle, named after the region of the
membranous labyrinth where they are located.
cells (outer phalangeal cells) that likewise
prevent their bases from touching the basal
lamina. Their stereocilia, about 100per cell,are
arranged in three or more rows that are in the
form ofa V or W.The rows ofstereocilia are also
of ascending height, from the inner to the outer
aspect of the cell. The stereocilia are embedded
in the tectorial membrane, a gelatinous
sheet of glycoprotein that moves in response
to pressure variations in the perilymph-filled
scalae tympani and vestibuli.
External to the outer phalangeal cells and
internal to the inner phalangeal cells are several
other types of supporting cells.
The hair cells in the organ of Corti are
stimulated by sound in the following manner.
When a sound wave amplified by the ossicles
in the middle ear reaches the foot plate of the
stapes, the oval window gets deflected inwards
and compresses the perilymph in the scala
vestibuli. This generates a compression wave
that travels upward along the coils of the scala
vestibuli to the helicotrema, then downward
along the coils of the scala tympani. This
compression wave produces a shearing force
that displaces the stereocilia of the hair cells
in the organ of Corti. The hair cells which are
mechanoreceptors then respond by releasing
neurotransmitters into the terminations of
sensory axons.
The round window, meanwhile, serves as
a pressure-relief vent by bulging outward into
the cavity of the middle ear with every inward
deflection of the oval window.
iflJJ :::.STE8!\N& CCNZ/\U:S TEXTBOOK OF HISTOLOGY
Vestibular Apparatus
The vestibular apparatus, as previously
stated, consists of the utricle, saccule, and the
three semicircular ducts.
The semicircular ducts have the general
shape-but occupy only about a quarter of the
cavity-of their corresponding semicircular
canal (i.e., anterior, posterior and lateral). Both
ends of each semicircular duct are continuous
with the utricle.
" The utricle is an irregularly-shaped
elongated pouch that occupies the
posterosuperior region of the vestibule. It is
connected to the saccule by a Y-shaped duct,
the utriculosaccular duct. One arm of this
duct, the endolymphatic duct, is lodged in
the vestibular aqueduct. The endolymphatic
duct has an expanded blind end called
endolymphatic sac that bulges into the
subdural space on the posterior surface of the
petrous part of the temporal bone.
The saccule is spherical and smaller than
the utricle. It lies on the anteroinferior region
of the vestibule. As already mentioned, it is
connected to the utricle by the utriculosaccular
duct and to the cochlear duct by the ductus
reuniens.
The wall of the utricle, saccule, and
semicircular ducts consists of an epithelium
and an underlying fibrous connective tissue.
The epithelium is simple squamous or cuboidal,
except in the endolymphatic sac and in the
five areas where the receptors for the sense of
balance are located, i.e., two maculae and three
cristae. The maculae are in the utricle and the
saccule (macula of the utricle and macula
of the saccule, respectively) while the cristae
(cristae ampullaris) are in each of the ampullae
of the semicircular ducts.
Maculae
The maculae are small regions of the
epithelium of the utricle and saccule. The
macula of the utricle is somewhat triangular in
surface view and is about 2.8 mm long and 2.2
mm wide. In contrast, the macula of the saccule
is an oval structure that is about 2.6 mm at its
widest, and 1.2 mm at its narrowest, diameter.
The maculae are structurally
\
similar. They
consist of a columnar epithelium that has two
basic types of cells: hair cells (sensory cells)
1
and supporting cells. _
The supporting cells are tall cells that have
a basally-locatednucleus and short microvilli on
their apical surfaces. At the edge of the macula,
" shaped. The basal two-thirds of each type I hair
Fig. XX-7. Macula of the Utricle (region at tips
of arrows). The macula is a small area of the
epithelium that consists of a columnar epithelium
(not very apparent in the photomicrograph)
made up of two types of cells: hair (sensory)
cells and supporting cells. The hair cells have
stereocilia and a single non-motile cilium which
are embedded in gelatinous material (otolithic
membrane). Found on the membrane's surface
are calcium carbonate crystals referred to as
otoliths. Ear, H&E x200.
they gradually flatten out to merge with the
simple squamous or cuboidal epithelium that
lines the rest of the utricle and saccule.
The hair cells, on the other hand, are
interspersed among the supporting cells.
Their nuclei are more centrally-located than
those of the supporting. cells. They have been
shown by EM to consist of two types: type I
and type II. The apical surfaces of both types
of hair cells are covered by 40-80 stereocilia
that, like those in the hair cells of the organ of
Corti, are graded in length. In addition, the
apical surfaces of the hair cells are provided
with a single non-motile cilium (kinocilium).
The stereocilia and cilium are embedded
in a plaque-like layer of gelatinous material
called otolithic membrane (statoconial
membrane) made up of glycoproteins and
is probably secreted by the supporting cells.
Scattered on the surface of the otolithic
membrane are calcium carbonate crystals
called otoliths (otoconia, statoliths).
Type I hair cells (goblet cells) are cup-
cell fits into and synapses with the cup-shaped
termination of the sensory axon of a bipolar
sensory neuron. Axonal terminations of motor
neurons (efferent neurons) synapse with the
sensory axon that surround each hair cell, but
not directly with the hair cell.
Type II hair cells (columnar cells) are
more slender than type I hair cells. They
synapse directly,but only at their bases,with the
terminations of two types of neurons: bipolar
sensory neurons and motor neurons (efferent
neurons).
The maculae detect the orientation of
the head with respect to gravity. With each
movement of the head, the otoliths create a
gravitational pull on the otolithic membrane
that causes it to move. Movement of the
otolithic membrane bends the stereocilia and
kinocilium, which stimulates the hair cells and
causethem to release neurotransmitters into the
terminations of the sensory axons with which
they are in contact.
EAR 'fiI

semicircular
dud
Cristae Ampullaris
Each crista ampullaris rests on a transverse
ridge of supporting connective tissue that
projects into the cavity of the ampulla of its
semicircular duct. This transverse ridge is
oriented perpendicular to the long axis of its
duct.
The cristae have many similarities with
the maculae. They consist of two types of cells:
supporting cells and hair cells (sensory cells).
The supporting cells are also columnar and
gradually flatten out in the edge of the crista to
merge with the simple squamous or cuboidal
cells that line the rest of the duct. The hair cells
are likewise of two types: type I and type II.
The hair cells resemble their corresponding
types in the maculae with respect to morphology
and the synapses that they form with the nerve
terminations. Hence, both types are provided
with stereocilia of graded height and a non-
motile kino cilium embedded in glycoprotein
material. The cristae differ from the maculae
in that the gelatinous glycoprotein material,
called cupola, which embeds the stereocilia and
kino cilium of hair cells in the cristae, is cone-
shaped and has no otoliths.
The cristae detect angular acceleration and
deceleration of the head. Angular movements of
the head cause a counterflow of endolymph in
each semicircular canal, moving the cupola and
ESTE8f.\~~
& (jONZALES' TEXTBOOK OF HISTOLOGY
Fig. XX-B. Crista Ampullaris.
Note that the crista ampullaris
rests on a ridge of connective
tissue which projects into the
cavity of the ampulla of the
semicircular canal. A crista
is very similar to a macula
in structure. It consists of a
columnar epithelium that has
two types of cells: hair (sensory
cells) and supporting cells. The
hair cells also have stereocilia
and a non-motile cilium
embedded in a glycoprotein
material. In the crista, however,
this material has no otoliths, is
cone-shaped, and is referred
to as cupola. Ear. H&E x100.
bending the stereocilia and kino cilium. This
causes the hair cells to release neurotransmitters
into the terminations of the sensory axons.
Innervation of the Maculae and Cristae
The bipolar sensory neurons whose sensory
axons (i.e., dendrites) are in contact with the
hair cells in the maculae and cristae comprise
the vestibular ganglion (Scarpa's ganglion),
located in the internal acoustic meatus.
Like those in the organ of Corti, aside
from sensory endings, the hair cells in the
maculae and cristae are supplied with axonal
terminations by motor (efferent) neurons whose
cell bodies are in the brain stem.
The axons of the bipolar sensory neurons
and efferent neurons form the vestibular
division of the vestibulocochlear nerve (eN
VIII).
Endolymphatic Sac
The epithelium of the endolymphatic sac is
atypical for the membranous labyrinth because
it is simple columnar rather than squamous
or cuboidal. The former also rests on a highly
vascular connective tissue. This epithelium
is probably the site for the absorption of
endolymph which, in the vestibular apparatus,
is produced by some of the supporting cells in
the utricle and semicircular ducts.