UNIT I5 MORPHOGENESIS AND TISSUE

ORGANISATION

Structure
Introduction
Objectives
Morphogenetic hocesses
Types of Morphogenetic Processes
Modcs of Cell Movement
The Role of Cytoskeletal Srructures Cell Movement
Adhesion of Cells to Extracellular Matrix
Morphogenesis of an Ec todermal Derivative
Neurulation in Amphibians
Neurulation in Chick
Mechanisms of Neural Plate Formation
Morphogenesis of Mesodermal Dcrivatives
Development of Heart in Amphibians
Development of Hcart in Chick
Development of Blood Cells
Morphogenesis of Endodermal Dcrivatives
Origin of Endodermal Organs
Origin and Migration of Primordial Germ Cells (PGC) in Frog
Origin and Migration of PGC in Chick and Mammals
Summary
Terminal Questions
Answers

15.1 INTRODUCTION

In the previous unit you have learnt about different patterns of clcavagc, mechanism of Morphogencsis is a Grcck
cleavage and how a single layered blastula gets converted into a gastrula with lhrce word meaning generation or
gem layers. In this unit, we shall discuss the postgastrulation changes which i) result in organisation of form.
the formation of definite body form by rearrangement of cells of the embryo, and ii) the
differentiation of organs and organ systems from thc germ layers - processes which are
collectively known as morphogenesis.
Further, in this unit, you will also lcarn how dilcercnt typcs of morphogenetic processes
lead to the formation of ectodermal, mesodermal and endodcrmal derivatives. It would
be interesting to know that there is a constancy about the gclm layers and the organs
derived fiom them, irrespctive of whcthcr the organism is a fish, a frog, a chick or a
mammal. The morphogenesis of certain structures of frog and chick will be discussed in
detail in his unit.

Objectives

Afia you have studied the unit, you should be able to:
list the various types of morphogenctic processcs and movcmcnts of cells,
e describe the n~orphogeneticprocesses that lead to the formation of neural tube in
amphibians and chick.
e explain the formation of heart as a mcsodcrmal dcrivative.
e discuss that the germ cells do not arisc from the gonad itsclr, but the precursors of
germ cells, the primordial germ cells, originate in chc endodcrm and migrate into
developing gonads.
Animal Development I
MORPHOGENETIC PROCESSES

In unit 14, we discussed the process by which a fertilised egg assumes the shape of a
ball or a sheet of cells around a cavity, called blastula. Subsequently, the cells in a
blastula actively rearrange themselves and take up positions in the gastrula from which
they give rise to adult organs. Essentially, there are three layers of cells, the ectoderm,
endoderm and mesoderm. Ectodermal cells will give rise to skin and its derivatives and
central nervous system. The cells that are aligilcd in the interior of the embryo, the
endodermal cells, will develop into alimentary canal and its derivatives; and the cells
which lie in between ectoderm and endoderm of a gastrula, the mesodermal cells, will
give rise to muscles, skeleton, connective tissue, urinogenital system etc. (Table 15.1).

Table 15.1 Major Derivatives of the Three Germ Layers In Vertebrates

Ectoderm
Nervous tissue
Sense organs
Epidermis of skin and derivatives

Mesoderm
Dennis of skin
Skeleton
Muscle
Circulatory system
Excretory system
Reproductive system (except germ cells)
Connective tissue .
Endoderm
Digestive system linings
Digestive glands
Lung and respiratory tract linings
Primordial germ cells
--
It should be obvious to you that a single cell, the fcstilised egg, gives rise to various
structures that we mentioned. Such a generation of cellular diversity is called
differentiation. The differentiation is a broad term which includes i) cytodifferentiation,
ii) histodifferentiation and iii) development of shapes of organs. The process of
differentiation which results in the formation of diverse organs and tissues involves
various types of cell movements termed as morphogenetic processes. The morphogenetic
processes are responsible for rearrangements of cell positions relative to each other.
Furlher there is the shaping and realignment of individual cells as well. The various
morphogenetic processes can be .wid to belong to eilher one of the six following
processcs. i) the direction and amount of cell divisions, ii) cell shape changes, iii) cell
migration, iv) cell growth, v) cell death and vi) changes in cell membrane and
extracellular matrix.

15.2.1 Types of Morphogenetic Processes

During early changes in the embryonic form and in the formation of organ rudiments.
the epithelial cells undergo a variety of folding and spreading movements. the processes
which shape the embryo. Transformation in the epithelial cells due to morphogenetic
processes may involve either the folding or branching or spreading of epithelial sheets.
The morphogenetic movements which bring about the required transformation are as
follows:
Any change in the epithelial sheet is preceded by a thickening of epithelium known
as palisading. Palisading occurs due to the elongation of cells (Fig. 15.1) and can
 be seen in the formation of neural plate and ectodermal laco odes such as lens. ear
and nasal rudiments.

15.1: Elongatlon of epithelial cells called palisading, a process that precedes any
morphogenetlc event In epithelial cells

Epithelium can be folded either inward or outward. When the epithelium bends .
outwards from the surface of the.embryo it is called evagination. On the other hand
when the folding is inwards, into the embryo or into a cavity, the phenomenon is
known as invagination.
Folding along a line will give rise to a groove (Fig. 15.2). The formation of aavrl
tube and laryngo-tracheal tube are examples of such a folding process.

Fig. 15.2: Formation of a groove (such as neural groove) by foldlng dong a llne of
thickened eplthelium.

ii) The formation of lens vesicle or otic vesicle from their respective'thickenings
illustrate inpocketing or infolding of the epithelium to form pouches (Fig. 15.3).

153: The tnpocketlng or lnfoldlng of eplthelium to form a pouch as in the case d

optic cup. /

Folds and pouches undergo modification to form branched structures. The formation
of various glands depends on the fold or cleft appearing at the epithelial outpocket
(Fig. 15.4).

Fig. 15.4: The branching of pouches as In the case of the formation of glands

Folding or bending of a sheit of cells may result in changes in the shape of the cell
within an epithelium. For instance, the narrowing of columnar epithelial cells at the
apical end results in the formation of pyramidal cells (Fig. 15.5). T'xis in turn results
Animnl Development I apical end results in the formation of pyramidal cells (Fig. 15.5). This in t~unresults
in the differences in the surface area on the two ends of epithelium and the bending
of the entire. sheet. Epithelial cells
/ 7

Fig. 15.5: Namwing of the columnar epithelial cells and the formation of pyramidal cells.

You may recall that while discussing gastrulation in amphibians, we described a

process called epiboly. In this process, there is a spreading of cells. The prospective
ectodem spreads to cover the embryo, Such spreading of cells also occurs at other
times during development. For example, during neurulation the epidermal ectodenn
(prospective skin) spreads to cover the area left vacant by the convergence of neural
epithelium toward dorsal midline. Spreading may be accompanied by a change in
cell shape such as thinning and flattening of individual cells (Fig. 15.6).

Neural ectoderm
*
Presumptive epidermis

Fig. 15.6: The process of flattening and spreading of epitheliak celk as it occurs during
neurulation process and future epidermis formation.

m Also, individual cells such as mesenchymal cells or primordial germ cells detach
froni major cell layers and migrate to new locations where they develop into
struchlres that they are programmed to develop.
In addition to various types of cell movbments that are involved in morphogenesis,
selective death of cells plays an imporrant role in shaping various structures of
developing embryo. Brain, limb and palate are the structures where cell death is
witnessed in certain regions during development Fig. 15.7 shows the necrotic region
or regions of cell death between developing digits of a chick that would ultimately
result in separate digits.

15.2.2 Modes of Cell Movement

In the previous subsection, we described the various types of morphogenetic processes.
Such processes are indicative of mobile nature of embryonic cells, and cell movement is
basic to morphogenetic process. In this subsection, we shall discuss the changes in the
shape of cells during morphogenetic process, as well as the factors that guide the cells
to their location.
 Morphogenesls and Tissue
Organlsation

Fig. 15.7: Necrotic zones in chick leg at various stages of development. Shaded areas are
necrotic zones where cells die at specific times during development. ANZ,
anterior necrotlc zone, mrZ- lnterdlgltal necrotlc zone, PNZ - posterior .
necrotic zone.

Fibroblasts which give risc to connective tissue have becn studied extensively to
understand the mechanism of cell mobility. The movcmcnt of fibroblasts occurs in two
phases: i) An adhesion phase in which the ccll stretches itself to the limits of its plasma
membrane and ii) a detachment phase in which the hindcr part of the ccll is pulled
forward. The cells propel themselves forward by generating regions that are quite thin
and fan shaped. Such regions of the cell are called the lamellae (Fig. 15.8). The lamella
bears a very thin and flat extension called lamellipodium. When the lamellipodium
protrudes foward, part of it gets detached from the substratum and folds back on itself,
thus giving a ruffled appearance.

Fig. 15.8: A migrating fibroblast. The cell moves In the direction In which the fan shaped
lamella with lamellipodla Ls formed.

The leading edges of such moving cells are termed as ruflled lamellipodia. The
larnellipodia are continued to be produced at the leading edge till the cell gets larger and
larger, and the hind end detaches itself from the substratum.
Since the cells move to and reach specific positions, we have to ask, what are the
mechanisms that direct the movement of cells to specific location in the embryo? It is
suggested that different mechanisms may be responsible for the guided moyement of
cells. They are a) chemotaxis b) haptotaxis c) galvanotaxis and d) contact guidance.
We shall briefly look into each one of these processes.
a) Chemotaxis: Chemotaxis refers to me directed movement of cells in response to a
concentration gradient of a chemical factor in solution. An example of chemotaxis
during morphogenesis of the embryo is the migration of embryonic lymphocytes
from bone marrow to embryonic thymus. The compound responsible for the directed
Animal Development I movement of cells is shown to be a heat-stable pcptide with a molecular weight of
1000 to 4000 daltons. The peptide is probably produccd by the thymus. The
gradient of the peptide would be highest in the thymus towards which the cells
move.
b) Haptotaxis: Haptotaxis refers to the directed movement of cells in response to a
concentration gradient of an adhesive molecule that may be present in the extra
cellular maxtrix. The adhesive material is not in solution. The cell would constantly
make or break adhesions with such molecules and move from the region of low
concentration of the molecule IO the place of its higher concentration. Poole and
Sternberg (1982) provided evidence that pronephric duct cclls in salamander
embryos move under the regulation of haptotaxis. The pronephri~duct rudiment
separates from the dorsal mesoderm as a solid cord of cells and it is at first seen
near the head of the embryo. With further development of embryo, this rudiment
ultimately elongates towards the cloaca where urine is excreted. Studies have shown
that the enzyme alkaline phosphatase may be the adhesive molecule that regulates
the migration of propheric duct rudiment.
c) Calvanotaxis: Galvanotaxis refers to the movement of cells in response to a
potential difference between cells. It is suggested that thcre are voltage differences
between embryonic regions that could play a significant role in morphogenesis.
Minute electric fields of the order of 10 to 100 mvlmm appcar to be sufficient to
alter the direction of nerve growth. Nuccitelli and Erickson (1984) showed that
embryonic chick fibroblasts migrateloward negative pole when cultured in small
steady electrical field. Early chick embryos, regenerating blastema of amphibians
and moth ovaries where transportation of yolk proteins occurs, arc some of the
structures where large electric currents have been detcctcd.
d) Contact guidance: Besides chemical or ionic factors that regulate cell movement in
developing embryos, physical factors also appear to play a role in morphogenetic
processes. A phenomenon called contact guidance influences cell movements in
cultures. The cell is influenced by the physical surface over which it passes and is
directed towards its location (Fig. 15.9). Weiss (1934) demonstrated that cells can
detect discontinuties in their substratum and migrate after aligning themselves along
such features as fibres. They could even move along scratches at the bottom of
petridish. In a recent study Harris (1980) showed that fibroblasts cultured on
silicone rubber or collagen change the shape of the substrate by the stress generated
by them, and move along such changed substrate fields. Contact guidance is
assumed to be the cause for the migration of mesenchymal cells inside the fish fin.

Fig. 159: The mammalian cells shown alignhg themselves on the grooved s u r f a m

15.2.3 The Role of Cytoskeletal Structures in Cell Movement

The term cytoskeleton refers collectively to a complex array of fibres in the cytoplasm
that mediate changes in cell shape and produce cell movements in embryos. The system
consists of three types of fibres i) microtubules ii) microfilaments and iii) intermediate
fwents.
i) Microtubules: All animal cells have microtubules as vital parts of basic cellular
structure and machinary (refer w Unit 3 of Block 1 of LSE-01; Cell Biology
 Course). Microtubules are hollow cylinderical rods and are 25 nm in diameter. Each Morphogenesis and Tissue
microtubule is formed of 13 rows of solid protofilaments which run parallel to the Organisation
long axis of microtubule. Each protofilament is a protein dimer composed of onea
and one tubulin chain. The protofilaments are assembled concomittantly side by
side to produce a hollow microtubule. Microtubules have a mechanical role in the
elongation or palisading of epithelial cells.

ii) Microfilaments: Microfilaments are also present in all animal cells. They are found
singly throughout the cytoplasm organised as a meshwork and measure 6 nm in
diameter. 'Ibese are the polymers of the contractile protein actin. F actin or
filamentous actin is the name gikn to the assembled polymers of actin. In
subsection 152.1, you have learnt that folding of cells brings about changes in the
cell shape within the cytoplasm and the apical surfaces of cells bccome narrower.
The narrowing of apical surfaces is brought about by contraction of microfilaments
made up of actin.
iii) Intermediate filaments: These filaments are intermediate in size between
microtubules and microfilaments and are 10.nm in diameter. Five classes of
intermediate filaments are known i) keratin filaments found in epithelial cells of
ectoderm and endodcrm ii) vimentin filaments found in mesoderm dcrived tissues
such as bone and cartilage iii) nerve filaments found in nerve cells iv) glial
filaments found in glial cells v) desmin filaments found in all types of muscle
tissues. The functions of different types of intermediate filaments are not clcar. It is
proposed that they may act with cellular organelles in organising and maintaining
their 3-dimensional structure.

15.2.4 Adhesion of Cells to Extracellular Cell Matrix

In subsection 15.2.2, you have learnt that the cells in ordcr to move or change shape,
adhcre to each other or to h e substratum in their environment. Generally the cells
adhere to complex environmental surfaces formed of molecules found in extracellular
mauix. Such molecules are known as extracellular matrix molecules or ECM
molecules. These molecules are secreted into spaces between cells and assemble into a
meshwork. Some molecules can form a dense sheet on the basal surface of epithelial
cells called basal lamina. The ECM molecules are capable of self-aggregation or self-
assembly and hence are able to form the extracellular matrix. Protcoglycans, collagens
and certain other glycoproteins belong to the category of ECM molecules. The binding
of cells to extracellular matrix is mcdiatcd by receptor proteins located in the plasma
membrane. Receptor proteins that mediate adhesion to several ECM molecules have also
been isolated.
Here, we end our discussion on certain of the general aspects of morphogenesis. Before
we start describing the morphogcncsis of sptcific structures from their respective germ
rr- layers, you may answer the following self-assessment questions.

I
SAQ 1
a. Define the tcrms i) differentialion and ii) morphogenesis.

b. Match the items in A with the ones found in B.

A B

i) Thickening of epithelium a) invagination

prior to the changes in
epithelial sheet
ii) Bending of the epithelium b) cell death
away from the surface of
Lhe embryo
Anlmal Development I iii) Folding of the epithelium c) epiboly
that results in the infolding
into the embryo
iv) The spreading of one layer d) palisading .
of germ cells over the other
v) Removal of specific cells by e) cvagination
necrotic process during
development

c. Fill in the blanks with suitable words.

i) The directed movement of cells in response to a concentration gradient of a
chemical factor in solution is known as
ii) refers to the movement of cells along a concentration gradient
of an adhesive molecule present in the extracellular matrix.
iii) Movement of eukaryotic cells in response to a potential difference between
them is referred to as
iv) The movement of cells influenced by the physical surface over which they
pass is termed as
v) The complex array of fibres. in cytoplasm that mediates changes in cell shape
and promotes cell movement in embryos is called

15.3 MORPHOGENESIS OF AN ECTODERMAL

DERIVATIVE

During the development of vertebrate bofy the different regions in the three germ layers
of the gastrula become segregated from each other to form the rudiments of future
organs and tissues. In this section we shall discuss the partitioning of ect$erm as well
as the formation and inward displacement of neural tube in frog and chick. The process
is called neurdlation. We have chosen to discuss the neurulation process bccause the
external morphology of developing embryo at this stage is dominated by the developing
nervous system and the embryo itself is known as neurula at this stage. We shall begin
with the separation of ectoderm into subpopulations of cells, 'each of which will develop
into distinct ectodermal organ. The ectodermal layer essentially separates into i)
epidermal ectoderm from which the skin is formed, ii) neural ectoderm which gives
rise to central nervous system and iii) neural crest which gives rise to a portion of
peripheral nervous system and a variety of othcr tissues.
15.3.1 Neurulation in Amphibians
The first step in the neurulation process is the flattening and thickening of dorsal
ectoderm to form neural plate (Fig. 15.10a). The plate of cells differs from the rest of
ecotoderm cells in that they have changed shape and appear more columnar. The edges
of the neural plate then rise above along the rest of the neural plate to form neural folds
(Fig. 15.10b) . The neural folds are now found along the two flanks of a central
depression called neural groove (Fig. 15.10~).The neural groove extends along the
entire middorsal line of the embryo. Eventually the neural folds meet in the middle
above the deepening neural groove and fuse to form the neural tube (Fig. 15. 10d. e).
At the time of rolling of neural plate to form the neural tube, the neuroepithelial cells at
the lateral folds of the neural plate develop constrictions at their apical edges. This
changes the shape of columnar eilhelial cells inlo pyramidal cones. As the surface area
of apical end of neural epithelial cells becomes smaller relative to the basal surface, the
rolling up of the neural plate and formation of neural tube takes place. With the fusion
of neural folds along the middorsal line, the neural ectoderm separates from epidermal
ectoderm which now covers the neural tube and completely surrounds the embryo. The
change in the shape of cells in the neural tube results in the regionalisation of neural
tube. In the cephalic end, i.e. the end which will differentiate into brain, the wall of the
tube is broad and thick and a series of swellings and constrictions develop that define
the vpious divisions of brain. Caudally, towards the posterior end, the tube is simple Morphogehesls and Tissue
Organisation
and narrow extending into the tail.

Neural plate Neural creast

A

Epidermis

Neural groove

Fig. 15.10: Stages in neurulation In frog. A) thlckenlng of dorsal ectoderm to form neural
plate B) formation of neural folds and neural groove C) deepening of neural
groove D) fusion of neural folds and formation of a neural tube E) neural tube
has separated and is covered over by epidermis. Neural crest cells have
separated from neural folds.

With the formation of neural tube, a second population of cells separate from the neural
ectoderm and lie between the neural tube and epidermis. This group of cells is the
neural crest cells (Fig. 15.10e). The neural crest cells subsequently migrate into various
parts of body to give rise to a variety of tissues in different parts of the body.
15.3.2 Neurulation in Chick
Neurulation in chick is similar to that of amphibians but there are certain differences. In
amphibians, the formation of neural tube occurs simultaneously along the entire length
of the embryo. In birds, reptiles and mammals even as the neuruhtion has begun in the
anterior part of the embryo, the posteior region is still in the process of gastrulation. At
a time when the neural folds are just a b u t to form in the gdsterior region, in the
anterior region the neural folds have already started fusing forminr! the neural tube
(Fig. 15.11)

Fk. 15.11: Dorsal view of a chick embryo of 25-26 hours wlth 5 pairs of somites Neural
folds approaching each other fuse and form neural tube in the cephalic region;
but In the posterior region gastrulation 1,s still occurring and primitive streak
and neural folds are yet to form.
 h i m 4 Development I

Fig. 15.12: Stages in the neurulation of chick. a) Neural plate formation (b, r, d) The
bending of plate In three locations (Lndlcated by arrows), Just above the
notochord (N) and on each side of the neural plate, Just below the tlps of neural
folds.
We described two important changes in cell shape while discussing neurulaeon in
amphibians. Such changes occur in chick embryo as well. One relates to changes in
neural ectoderm cells from cuboidal to columnar shape that results in narrowing and
thickening of neural plate. The second relates to narrowing of the apical ends of some
neural epithelial cells when the neural plate rolls up to form neural tube (Fig. 15.12b. c,
dl.

15.3.3 Mechanisms of Neural Plate Formation

You are now farnilar with the concept that the neurulation comprises of two processes i)
the neural plate formaton and ii) the neural tube formation. The mechanisms remnsible
for these two processes are not very clear.
The shaping of neural plate from an oval to a long, narrow plate is believed to occur by
'
the elongation of neural epithelial cells and the accompanying apical shrinkage.
Although it was earlier believed that microtubules (refer to subsection 15.2.3) may be
involved in the elongation process, evidence is inconclusive to suggest a role for
microtubules in the elongation process. The role of microtubule appears to be important
in stabilising the elongated state of the cells rather than in the elongation itself.
Several hypothesis have been put forward to explain the bending of neural plate to form
neural tube. The more acceptable one is that during the rolling of neural plate, the apical
surface of neural epithelial.cells contract at the site of bending because of the
contraction of actin microfilaments (refer to section 15.2.3).
Before we proceed to discuss the mesodermal derivatives attempt the following SAQ.
SAQ 2
State wether the following statements are true or false.
i) During post-gastrulation development, different germ layers of the gastrula become
segregated from each other to form tissues and organs.
ii) The term neurulation refers to the partitioning of ectoderm, and the formation and
inward displacement of neural tube.
iii) Neurulation results in the separation of ectoderm into epidermal ectoderm. neural
ectoderm and neural crest cells.

I iv) Neurulation process in chick is entirely different from that of amphibians.

v) During neural plate formation, the neuroepithelial cells change in shape from
columnar to pyramidal ones.
 vi) Microtubules play a signif?ant role during the rolling up of the neural plate into Morphogenesls and T i m
neural tube. Organbation

vii) In chick the neurulation process occurs simultaneously through out the length of the
. embryo.

15.4 MORPHOGENESIS OF MESODERMAL

DERIVATIVES
In this section, you will study the early development of an organ derived from
mesodem. In fact, all the organs that lie between ectoderm and endoderm tissues arise
from mesoderm. In the neurula stage of the embryo, the mesoderm cells are arranged in
five distinct regions. The five regions of the mesoderm and the organs derived from
them are as follow?:
I
I
Chordamesoderm: Chor&mes&rm separates as a middorsal strip from the rest of
the mesodermal tissue and establishes the body,axis of the embryo. The anterior
part gives rise to head mesoderm and the remaining part to notochord (Fig. 15.13B).
I
Dorsal mesoderm or paraxial mesoderm: Tissues developing from this region will
be in the back of the embryo, on either side of spinal cord. It segments into blocks
I of tissues called somites. (Fig. 15.13C. D). Connective tissues and associated
structures such as muscles, cartilage and dcrmis arise from this region.
Intermediate mesoderm: Intermediate mesoderm is located as a thin stalk
connecting paraxial mesoderm with the rest of the mesodermal sheet The
urinogentital system arises from intermediate mesoderm (Fig. 15.13C. D).

D '

Lateral plate,

Flg. 15.13: Stages in the development of mesoderm shown In transverse sectlorn of chick
embryo at trunk level.

I Lateral plate mesoderm (Fig. 15.13D): Lateral plate mesoderm is a sheet of

loosely connected cells on either side of gut (Fig. 15.13C). This loose sheet of
epithelial cells splits into two layers, the somatic mesoderm which becomes closely
associated with ectodenn and the splanchnic mesoderm which becomes closely
, Animal Development I associatcd with endodcrm. Thc spacc bctwccn the two rcgions of mcsoderm is the
future coelomic cavity (Fig. 15.13D). Latcral plate mcsodcrm givcs rise to heart.
blood vessels and blood cells and the lining of the body cavities. All the
components of limb except musclcs are derived from lateral plate mesoderm.
Head mesoderm The hcad rnesodcrm located in the hcad region will give rise to
head muscles.
We shall now describe the developmcnt of one of h e organ rudimcnts dcrivcd from
mcsodcrm. It is a common pncticc to dcscribe thc dcvelopmcnt of limb rudiment as an
example of an organ differentiating from latcral mcsodcrm. In fact, all the processes of
developmcnt can be seen in the formation of a limb and the major featurcs of limb
development are common to all vcrtcbratcs. Since the development of limb is discussed
in detail in Unit 17 of this block, in this section we shall dcscribe the developmcnt of
two other mesoderm dcrivativcs, the hcart and thc blood cells. The circulatory system is
the first functional system in the dcvcloping embryo and the hcart is thc first functional
organ. The development of circulatory systcm with heart, blood cclls and a nctwork of
blood vessels is more complcx than the developmcnt of othcr systcrns. As examples, we
shall discuss the process of dcvclopmcnt of hcart in amphibians among lower vcrtcbratcs
and chick among the amniotcs. The differences beiwecn the two are largely dctcrmincd
by the differences in the amount of yolk in thcir eggs.

15.4.1 Development of Heart in Amphibians

The heart and the surrounding pcricardial cavity develop from lalcral mesoderm. After
gastrulation, the medodermal mantles continue to grow anteriorly and ventrally. At the
ventral ends of these mantlcs bclow the gut and in the space bctwccn thcm there occurs
proliferation of cclls (Fig. 15.14A) which initially form a cord (Fig. 15.14B) but
subsequently the cord of cells hollows out to form a tube (Fig. 15.14C). The tube made
of endothclial cclls is called endocardium, meaning the inner lining of heart. Similar
endothclial tubes are present anterior and posterior to endocardium and hcsc tubes will
become the maior blood vcsscls to and from hcart.

Rudiment of / endocardium
- endocardrum / D . .

mesocardium E Iuoe I - ,,.

Endocardium
Fig. 15.14: Cross section of frog embryo showing stages in the development of heart.

Once the endocardium is formcd, he rnantlcs of mcsodcrm meet and fuse in he middle,
ventral to the endocardium. The splanchnic mesodermal laycr thcn spreads dorsally on
either side of the endocardium ultimately surrounding the endocardium and joining
above it. The portion of splanchnic mcsodcrmal laycr surrounding the endocardium is
the prospcctivc muscle laycr of the hcart, the epimyocardium (Fig. 15.14D, E). The
regions of the mcsodcrm above and bclow the hcart, whcre Lhc fusion of the mcsodcrm
 of the two sidcs have faken place, become the mescnterics that suspend the heart in Morphogenesls and Tlssue
pcricardial cavity (Fig. 15.14E). Thc ventral mesentcry disappears later. The coelomic Urganlsation
cavity now becomes thc pericardial cavity and the somatic layer of mesoderm bccomcs
the lining of the poricardial cavity, Lhe pericardiuni.

15.4.2 Development of Heart in Chick

You have learnt earlicr that in amniotes bccause of the prescnce of a large mount of
yolk, the germ layers are found as flattcncd sheets on the yolk. In chick the heart
develops first as a pair of tubes and thcn the two tubes fuse to form a single tube.
During the head fold stage of embryo, the cells of the splanchnic mesoderm detach from
the epithclium and migrate to lic below the foregut endoderrn to form two groups of
cells. In each group, the cells coalesce and form a thin walled tube (Fig. 15.15A.B, C).
The two tubes then fuse in the midline below the gut to form the endocardium (Fig.
15.15C, D). As in amphibians, here also the regions of splanchnic mesoderm that lie
dorsal and ventral to endocardium fusc forming the myocardium (Fig. 15.1fC, d). The
dcvclopment of chick heart fakes place in the anterior-posterior direcdon w h i ~ hmeans

Ectoderm Neural groove Neural canal

Pericardial
region of
coelom
Splanchnic - ium
re; of \ mesoderm
emerging Open gut caudal to
I endocardial anterior intestinal portal 1 Cardiac 1'
tubes 1
1

c Foregut
D Neural tube

Cardiac
jelly
Myocardium --

Ventral mesocardiu
L-mesocardium (Disappearing) jelly
1
Fig. 15.15: Stapes in the development of heart of chlck from splanchnlc rne.wderm.

the fusion of the endocardia1 tubes takes place initially antcriorly and subsequcnlly
posteriorly (Fig. 15.16A to D). In a 33 hour old chick embryo, a tubular heart is
formcd but thc chambcrs havc not yet dcvclopcd. Thc hcart is already beating with the
blood entering the hcart at the posterior cnd which is the future atrium or auricle. The
blood is then pumped into ventricle from where it flows out through devcloping aortic
arches. Thc coelorn enlarges to form the pericardial cavity surrounding the heart and is
1inc.d wiQr pericardium.
The tubular heart of an early cmbryo develops into an adult heart around 120 hours of
embryonic life. The dcvelopmcnt of a four chambered heart from a tubular structure
with thc separation o'f pulmouory and systemic circulation dcpcnds on two significant
morphogcnctic events. i) The atrium is brought dorsal to the vcnuiclcs by the process of
looping and bending and ii) division of thc tube into chambcrs is achieved by the
formation of scpla. Figs. 15.17A to F show the process of looping that transforms the
tubular heart Into a chambered onc. The transformation appears Lo be an endogenous
property of the hcart and also lnvolves certain ccll shape changcs in the myocardial
epithelium. The dctails of the forces that bring about the transformation are not clearly
undcrslood.
I
I Animal Development I

Fig. 15.16: Stages in the development of the heatt In the chlck embryo s h o r n from the
ventral side. Note the antero-postelor fuslon of the paired cardlac prlmordle

15.4.3 Development of Blood Cells I

In this subsection we shall discuss the development of blood cells, mainly erythrocytes \
or red blood cells CRBCsl. Erythrocytes are the most numerous cell type in the blood
venrrlu aonue

fusion of primordia fusion of atrial primordia sinusvenosus

of ventricle 10 somites 14 somites
8 somites
aortae
ventral tmncus arteriosus
I

.
,.. p::
ventriculir ,
loop

fusion of primordia 48 hours 72 hours

of sinus venosus ,
21 somitts ----
Fit.
- 15.17: A) The fusion of the heart rudiments of the chick embryo (ventral views). The
heart rudiments conslst of two tubes which fuse to form a slngle tubular heart.
This fusion begins at the level of the ventricles (28 hours) and continues
progressively in a posterior direction. At 30 hours the paired rudiments of the
. atria begin to fuse, and by 33 hours a single atrium is formed. The sinus
venosus is still present a s two primordia at 34 hours. B) Form changes in the
heart of the chick embryo (dorsal vlew). At 40 hours the paired rudiments of the
sinus venosus are fusing and the ventricle becomes bent Into a loop. Further
bending of the ventricle at 48 hours places It in a position lateral to the atrium.
and at 72 hours the ventricle Is posterior to the atrium.
which also contains the different types of white blood cells or leucocytes including Morphogenesis and Tissue
Organisation
granulocytcs, monocytcs, platelets, plasma cells and lymphocytes. Our knowledge about
the developrr~e~ilof bload cells is derived mainly from studies on birds and mammals.
All types of blood cells have a limited life span. For example, a human RBC survives
for only about 120 days and in a healthy person trillions of RBCs and other blood cells
a're lost every day and replaced by continuous production of new cells in the bone
marrow from the haematopoietic (blood forming) stem cells.
A stem cell is an undifferentiated cell, capable of extensive proliferation that can
generate differentiated cells as well as more undiflcrentiated, embryonic cells of its own
tjpe. Thus a population of such embryonic stem cells is maintained even in the adult
that guarantees continuous replacement of difl'erentiated cells of specific tjlpe that are
dying and are lost throughout lire of the animal.
Experimental evidence indicates that in birds and mammals all types of blood cells are
-
derived ultimately irorn one type of stem cell callcd CFU M, L (myeloid and
lymphoid colony forming unit). This cell is pluripotent and generates both r 4 and
white blood cells in addition to itself.
However, CFU - M, L does not give rise to various blood cell types directly. Instead, in
addition to reproducing its own kind it produces two other types of stem cells callcd
CFU - S and CFU - L. These are also pluripotent but with lesser potentiality than that
of the parent CFU - M, L. Thus, white CFU - S can generate erythrocytes, granulocytes,
monocytes and platelets bur not lymphocytes; the CFU - L can give rise to only
lympl~ocytesand plasma cells.
In its turn, CFU - S generates, in addition to more of itself, five othcr types of stem
cclls (BPU - E, n a - CPC, GM - CFC and Meg - CFC). Each of these is capable of
generating differentiatcd cells of some specific type(s) in addition to replacing itself.
Each of these cells is therefore, called a committed stem cell. Among them the stem
cells RF'U - E (Blood forming unit - erythroid) are committed to the erythroid pathway
leading to the formation of erythrocytes only (Fig. 15.18).
By repeated divisions the BFU - E cell eventually produces a propulation of cells called
proerythroblasts. In mammals the proerythroblast cell passes through a series of stages
ultimately becoming a fully differentiated and functional erytl~rocyte(Figs. 15.18,
15.13). During this process many changes take place in the cell and each stage is
characterised by certain features of its own (Fig. 15.19).
Proerythroblast stage : Active RNA synthesis and proliferation
Erythroblast stage : Chromosomal condensation; beginning of
hemoglobin synthesis
Polychrornaoophilic stage : Increased synthesis and accumulation of
hemoglobin; decreased RNA synthesis
Orthochromatic stage : Nucleus completely inactivated; division no
longer possible
Rcticulocyte stage : Nucleus extruded; some hemoglobin synthesis
still occurring; cell enters blood stream
Erythrocyte : No more synthetic activity of any kind; cell is a
membranous bag filled with hemoglobin solution
Transformation of the progeny of BFU - E into procrythroblast cclls and the subsequcnt
process of their differentiation into erythrocytes occur under the influence of a hormone,
erytliropoietin, secreted in the'kidneys. If 0,supply is deficient, production of this
hormone is enhanced leading to increased number of BFU - E progeny transforming into
proerythroblasts. Ultimately the proerythroblasts are terminally dilerentiated into
functiorlal red blood cells.
In adult mammals the major site of the formation of blood cells is the bone marrow.
Experimental studies show that in embryos the f i s t pluripotcnt haematopoietic stem cells
(CFU - M, L) originate in the mesodermal blood islands of yolk sac. They reach and
colonize successively the liver, and then the spleen and bone marrow during later life of
the fetus. In bird embryos these master stem cells appear to originate in the yolk sac as
well as the blood vessels within the emrbyo proper.
Anlm'al Development I

Red blood cell Basophils Monocytes, Neutrophils Eosinophils Platelets Plasma Activated T cell
(Erythrocyte) macrophages ccll

Fig. 15.18: Schematlc diagram showlng the origin and developmental pathways of blood and
lymphold cells.

Differentiation of blood cells begins first in the yolk sac of the embryo, later in the fetal
liver and last injthe bone marrow. In mouse it bcgins in the yolk sac on 8th day, in the
fetal liver on 12th day and in the bone marrow it occurs from 16th day of gestation
onward. In the human fetus blood cell dilerentiation bcgins in the yolk sac on 9th day
and in the bone marrow aficr the first trimester.

Fig. 15.19: Diagrammatic representation of the stages in the differentiation of erythrocyte

(RBC). a) The mesenchymal pluripotent CFU - S which can give rise to more
-
than one type of stem cell. b) The hemocytoblast, (UFU E), the stem celi of
erythroid line. c) Prcxrythroblast d) Erythroblast e) Iblychromatophilic
erythroblast f) Orthoclirornatic erythroblast. g) Reticulocyte. Terminally
differentiated erytlirocyte stage (RBC) follows the reticulocyte stage.

SAQ 3
a) Match correctly the region of the mesoderm (A) with the organs derived from it
(B).
i) Intermediate mesoderm a) notochord
ii) Dorsal mesoderm b) heart and limb
iii) Head mesoderm
iv) Lateral plate mesoderm
c) rnusclcs, cartilage and dermis
d) muscles of the face
/
/'
v) Chordamesoderm e) urinogenital ducts ,/
b) Choose the correct answer from the alternatives provided.
7
/'

i) The fxst functional system in a developing embryo i s k o u s system,

ii)
circulatory system.
//
The heart and the pcricardial cavity devcloflrom dorsalPateral mesoderm.
,
iii) The proliferating 11s of mesoderm ultimately form the
heart tube called
iv) The epimyocardium rise to the muscle layers of the heart
differentiates from
v) In a 30 a tubular heart is formcd but the chambers are not

vi) The coeQf;;enlarges to form the chambers of thcheanlpericardial cavity.

hcart into a chambered one the future atria/
dorsal to h e atria/vcntriclcs by processes of looping and
bending.
h t a t e ' whether the following statements are true or false.
i) The development of chick h e k takes place in the postrior-anterior direction.
ii) Both in amphibians and chick the somatic mesodcrm that lie dorsal and
v e n d to endocardium fuse forming myocardium.
iii) In the tubular embryonic heart the blood enters at the posterior end, pumped
into the ventricle and flows out through the dcvcloping aortic arches.
iv) The transformation of tubular heart into a chambcrcd one dcpends exclusively
on cell shape changes in myocardial epithelium.
v) The oxygen supply to the developing embryos of chick is through lungs.
vi) The C N - S and C'FU - L are commilted cell's which develop into a specific
ccll type.

15.5 MORPHOGENESIS OF ENDODERMAL

DERIVATIVES

Thus far we have discussed the morphogeneis of organ rudiments derived from ectoderm
and mesoderm. In this section, we shall discuss about endcderm and its derivatives.
The thud and the innermost germ layer endodcrm mainly gives rise to gut tube and its
accessory organs, 'respiratory apparatus and primordial germ cells (PGC). The
development processes of endoderm includes long distance migration of germ cells,
prominent foldings and a number of evaginations from the digestive tube.

15.5.1 Origin of Endodermal Organs

Following major endodermal organs arise as evaginations of the digestive tube.
In pharyngeal region paired pouches bulge out laterally, meet ectcdermal
invaginations and form gill slits which in the higher vertebrates give rise to cords of
Animal Development I cells that form among other structures, the eustachian tube, thymus and
para thyroid gland.
From the floor of pharynx cords of endodcrmal cells pcnetrak ventrally into the
underlying mesoderm forming thyroid gland.
A midventral groove (laryngo - tracheal groove) from the floor of pharynx gives rise
to trachea and lungs.
Further, posteriorly at the level of the future duodenum, the other evaginations
iniiiqte the development of liver, pancreas and gall bladder.
The orgari that bud off from the pharynx and gut are not purely endodermal in
their final staic; mesodcrm invests them and supplies blood vessels; and a
framework of conricctive tissue shapes each organ into its definitive structure.
The primordial germ cel'!s that migrate from endoerm deserve special attention
because the gonads in which they are'dtimately found are mesodermal in origin.
In this section you will learn about the crigin of primordial germ cells and their
subsequent migration to spccific destination in vertebrates.

15.5.1 Origin and Migration of Primordial Germ Cells in Frog

In frog eggs the germplasm present as granular material is lasted in the cytoplasm near
the vegetal pole of the egg. During cleavage the germinal granules are found in the
endodermal blastomeres of this region (Fig. 15.20A). Later, the descendents of these
cells known as primordial germ cells (PGC) migrate dorsally around the posterior gut
region and enter into mesodermal region (Fig. 15.20B - E). The mesodermal region into
which PGC enter are the genital ridges (Fig. 15.20F). The gonads develop from these
ridges. In the developing gonad the PGC derived from endoderm divide and form gonial
cells which undergo meiosis to produce gametes.
A

PGC

Genital ridge

Fig. 15.20: Diagram showing the distribution of primordial germ cells (PGC) in the vegetal
blastomeres and thelr subsequent migration to the genital ridge in frog embryos.

At about the end of gastrulation the primordial gcrm cclls are found on the floor of
archenteron (gut endodcrm). Thc migrating primordial gcrm cclls of frog (Xenopus)
embryos lcave the floor of the gut, move laterally and are thcn found aligncd with
dorsal mesentery. The PGCs migrate along this mcscntery until they reach the
developing gonads. Such an alignment suggests that contact guidance (see section
15.2.2) directs the moverncnt of gcrm cclls to gonads. It is suggested that adhesion of
PGCs to the substratum is mediated by fibronectin produced by the mesentery cells. The
frog PGCs adhere with and align on tha fibronectin fibrils when they begin their
migration into gonads.

15.5.2 Origin and Migration PGCs in Amniotes

In birds and reptiles the primordial germ cclls originate in the epiblast (see marginal
r-arks), and move into underlying hypoblast (see marginal remarks) at the anterior
border of area pellucida of the primitive strcak stage embryo. This region is known as Morphogenesis and Tissue
Orgenisatiun
germinal crescent region. Here the PGCs appear larger than thc o~herembryonic cells
(Fig. 15.21). In the chick the PGCs can be identified cytochemically by their large
Epiblast is the layer of cells
glycogon content. During further development PGCs enter the space between hypoblast lying above the blastocoel
and epiblast and finally invade the developing blood vessels. They are carried passively in a discoidzl cmbryo such
in the blood stream through the blood vessels until they reach the the genital ridges. as chick embryo.
Form the genital ridge they migrate through the mosoderm cells to reach the dcveloping
gonads.
Hypoblast is the layer of
cells below the blastocoel
in a discoidal embryo.

Area
opaca

Flg. 15.21: Prlmltlve streak stage of chlck embryo showing prlrnwdlal germ cells tn the
crescent at the anterlor border between area peUuclda and area opaca.

From the genital ridge to the gonads the movement is mediated by chemotaxis (see
Section 15.2.2). Experimental studies have indicated that gonad tissues release a
diffusible chemotactic molecule that attracts and directs the movement of PGC to their
sites of location.
In mammals, the PGCs have high concentration of the enzyme, alkaline phospbatase and
can be distinguished from other embryonic cells by staining for this enzyme. They are
first seen in the yolk sac endoderm near the base of embryonic allantois (Fig. 15.22A).
Here, they break up into two streams, each of which migrates through the developing
gut into the dorsal mesentery and finally into the genital ridge of its respective side
(Fig. 15.22B). In mouse it has been shown that besides the active movements by
fdopodia and chemotaxis, contact guidance also plays a part in the movement of PGCs
Gentid ridge B
/

ridge

-
Fig. 1532: Orlgh and mlgretlon of mammallm prlmordlal germ cells A) Prlmordlal germ
celk first reeogaked In the yolk sac. B) Mlgretlon through gut end dorsally up
the dorsal mesentery Into the genltal ridge.

We end our discussion here on the morphogenetic processes and the morphogenesis of
specific organs from their germ layers. You may attempt the following SAQ to check
your unberstanding of this section of the unit
SAQ 4
Fill in the blanks with appropriate words.
 Animal Development 1 i) Endoderm gives rise to tube and its accessory organs,
apparatus and
ii) Gonads arise from the layer and primordial germ cells from 1
iii) The border between area pellucida and area opaca where the primordial germ cells
are first seen is known as
iv) In Xenoprcs embryos the migration of primordial germ cells to the gonads is
mediated by
v) Zn chick the migration of PGCs from genital ridges to gon@'is mediated by

vi) In mammals the migration of PGC from the site of their origin to the gonads is
mediated by and

SUMMARY
I In this unit you have learnt the meaning of the word morphogenesis, the different
types of morphogenetic processes and the mechanisms which mediate the cell
movements in vertebrate embryos.
We discussed the role of cytoskeletal components such as microtubules, I
microfilaments and intermediate filaments in cell movement during morphogenesis. i
You have also learnt about the nature of molecules to which embryonic cells adhere
when they undergo change in their shape in order to effect morphogenetic
movements.
In this unit we also discussed the morphogenesis of organ rudiments from three
i
germ layers of the embryo viz. the ectoderm, mesoderm and endoderm. We
discussed the newulation process which results in the separation of ectoderm into
e~idermalectoderm and neural ectoderm and the subsecruent separation of neural
&to&& into neural tube and neural crest cells. These *&cess& are described in
amphibians and in chick. Neurulation consists of, first the neural plate formation
and then the neural tube formation. The neural plate is formed by changes in the 1
shape of the cells by the cell elongation and accompanying apical shrinkage. The
rolling of neural plate into neural tube occurs due to changes in cell shape brought
about by the con~aclionof actin filaments.
We have listed the five distinct regions of mesoderm found in neurula stage and the
organs derived from them. We have discussed in detail the development of heart in
amphibians and chick as the derivatives of lateral mesoderm. The differentiation of
different blood cells from the pluripotent hematopoietic stem cell is also discussed.
Finally, the various organ rudiments derived from endoderm are listed. In
amphibians, chick as well as mammals, the gonads are derived from mesoderm but
thiprimordial germ cells (PGCs) arise fromkndodermal cells. From the site of their
origin the PGCs migrate to the genital ridge from where they find their way into
gonads to differentiate into gametes.

TERMINAL QUESTIONS
-

1. What are the six different processes that characterise morphogenesis in vertebrates?
..........................................................................................................

2. Explain briefly the following terms in relation to morphogenesis i) palisading

ii) invagination iii) evagination iv) cell death.
Describe the following types of movements exhibited by cells of a developing
embryo.
a) Chemotaxis b) Contact guidance

..........................................................................................................
Describe the process of neurulation in chick.

..........................................................................................................
Discuss the differentiation of an erythrocyte from a pluripotent CFU - S cell.

Describe briefly the origin and migration of PGC in chick embryos.

..........................................................................................................
..........................................................................................................
..........................................................................................................
..........................................................................................................
..........................................................................................................
..........................................................................................................
 iv) contact guidance Morphogcnesis and Tissue
Organisatlon
v) chemotaxis
,
,

vi) fdopodia, chemotaxis, contact guidance

TERMINAL QUESTIONS

1. i) direction and amount of cell divisions

ii) cell change shapes
iii) cell migration
iv) cell growth
v) cell death
vi) changes in the composition of cell membrane and extracellular matrix.
2. Refer to the text in subsection 15.2.1.
3. Refer to the text in subsection 15.2.2.
4. Refer to the text in subsection 15.3.2.
5. Refer to the text in subsection 15.4.3.
6. Refer to the text in subsection 15.5.2.