VETA 60.

VETERINARY DEVELOPMENTAL ANATOMY

MODULE 6
CONCEPTS & MECHANISM OF DEVELOPMENT
EMBRYONIC DUPLICATION AND TWINNING

LEARNING OBJECTIVES:
After the completion of the module, you will be able to:
1. Discuss the developmental processes involved during the growth and development of
organs and structures in the embryo;
2. Explain how these developmental processes are accomplished using cell signaling and
communication within the embryo;
3. Describe the origin and causes of embryonic duplication and twinning; and
4. Identify and classify and the different types of embryonic duplication and twinning.

Embryonic development encompasses all the processes whereby a single cell – the
fertilized egg or zygote – gives rise to first an embryo, then a fetus which at birth has the
capacity to adapt to postnatal life. These processes occur continuously but may be divided into
different periods. Thus, intrauterine development is often divided into an embryonic period,
where all major organ systems are established, and a fetal period, which consists primarily of
growth and organ refinement. Development of the organism does not stop with birth, however;
organs continue to grow and mature at least until puberty and many tissues need continuous
replenishment throughout life. Aging and death may therefore also be included the natural
developmental process of the organism.

CONCEPTS & MECHANISM OF DEVELOPMENT

Within the fertilized ovum lies the capability to form an entire organism. Individual cells
resulting from the first few divisions after fertilization retains this capability, thus cells are
described as totipotent. As development continues, the cells gradually lose the ability to form
all the types of cells that are found within an adult body and would be destined to become
specific types of cells (Figure 6.2).
Most early embryos pass through a stage called the “blastula or blastocyst stage” at which
they consist of a featureless ball or sheet of cells. The cells in the different regions needs to
become programmed to form the different body parts such as the head, the trunk, and tail. The
initial step usually involves regulatory molecules (i.e. determinants) deposited in particular
locations within the fertilized egg. The latter steps involve intercellular signaling events known
as embryonic induction, which lead to the upregulation of different combinations of
developmental control genes in each zone of cells.
These developmental processes are involved during the growth and development of the
organs and structures in the embryo and also the embryo as a whole.

GROWTH

Growth is simply defined as an increase in mass. This involves increase in the size of a
part or the entirety of the organism brought about by increase in the size (hypertrophy) and/or
number of cells (hyperplasia). This process continues even after the organism has hatched from
its egg or born. A striking feature of young embryos is the rapid growth of the head region. This
results in a relatively large head in the embryo and fetus. Later, when growth in the other parts
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of the body catches up, adult proportions are established through differential growth of more
caudal regions of the body and limbs. There are two major patterns of growth in animals. In
determinate growth, the body grows to a certain point that is characteristic of the species and
sex, then growth ceases. This is common in mammals. Indeterminate growth on the other
hand, is more common in ancestral vertebrates like fishes. Growth in these animals continue
throughout the lifespan although at a reduced rate in later life (e.g. age determination of fish by
examining the annual growth rings on scales).

MORPHOGENESIS

Morphogenesis is the mechanism by which tissues and organs are shaped. During
morphogenesis, structures such as tubes, sheets, and dense clumps of cells are formed in
response to differential rates of cell proliferation, changes in cell size and/or shape, cell fusions
and/or changes in cell adhesion properties.
Different types of morphogenesis occur in developing embryos, these are:
1. Epithelial expansion – spreading out of epithelia (Figure 6.1). May be through:
a. epiboly – extension by thinning of cells
b. intercalation – rows of cells (in adjacent layers) move between one another to
create an expanded epithelium but with thinner layer of cells
c. convergent extension – several layers fuse to expand the epithelium
2. Delamination - cells breaking free from the parent epithelium or splitting of epithelium
into thin layers
3. Ingression - process by which cells migrate from the surface layer of the embryo into
the interior as individual cells. (See Figure 6.3)
4. Mesenchymal aggregation – clumping together of loose cells to form a mass
5. Placode formation – thickening of surface epithelium
6. Invagination – inward movement or depression of epithelium
7. Branching
8. Vesicle formation – formation of a spherical structure. Usually formed initially either
by invagination or evagination
9. Evagination – outward bulging

Figure 6.1. Epithelial expansion and how it occurs in different forms.

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Figure 6.2. Differentiation of the derivatives of the zygote into the tissues of the body.
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Figure 6.3. Other types of morphogenesis: Invagination, Ingression and Involution

CYTODIFFERENTIATION

The adult mammalian body is composed of more than 230 different cell types, all
originating from a single cell, the fertilized egg or zygote. The process whereby specialized cell
types develop from less specialized is known as cell differentiation or cytodifferentiation.
Cytodifferentiation is a complex process wherein a cell or cell line attains and produces a stable
phenotype (e.g. maturation of sex cells) or simply mean the loss of pluripotency (ability to give
rise to different cell types). It is ultimately regulated through differential gene expression. Like
a stream running down the side of a mountain with its flow branching many times before it
reaches the bottom, so do embryonic cells differentiate from common origins to gradually form
specialized cell types. And just as a leaf dropped onto the stream follows one path only, so
does a particular cell follow a single line of differentiation. However, for both the leaf and the
cell, numerous decisions are taken along the way to the final destination (Figure 6.2). Hence,
cell differentiation during embryonic development involves many branch points where lineages
divide and sequential decisions on differentiation are taken. These decisions are at first
reversible (cell specification), but later become irreversible (cell determination).

PATTERNING

Patterning is the establishment of self-moving subsets of cells in proper relation to each

other and to other surrounding tissues (e.g. feathers and nails on skin, formation of teeth and
gums in the oral cavity, etc.). While differentiation gives rise to cells with specialized structure
and function, this process alone does not form an organism; the differentiated cells need to be
spatially organized in three dimensions and in well-defined relationships to each other. All
mammalian embryos tend to follow basic body plans providing craniocaudal, dorsoventral and
proximodistal axes.
Patterning is the consequence of regional gene expression which may be installed
through the action of gradients of signaling molecules. Target cells closer to the signaling
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source receive signals in a higher concentration than do those located more distally. Signaling
molecules working in this way are known as morphogens. The embryonic craniocaudal and
proximodistal body axes are patterned using more or less known morphogenetic fields. An
example of patterning by regional gene expression controlled by gradients of signaling
molecules is the formation of the neural tube.

APOPTOSIS

Apoptosis refers to genetically programmed death of unwanted cells. This process

occurs in many cell types during normal development. Cell death due to apoptosis is distinct
from necrosis which can occur as a result of acute non-specific injury to the cell. Different
apoptotic pathways exist in mammals and extracellular and intracellular stimuli can trigger this.
Caspases are examples of intracellular mediators of apoptosis. Once activated, they can cleave
proteins essential for the survival of the cell. Apoptosis also plays essential role in the
remodeling of interdigital tissue.

The processes enumerated above are

genetically programmed. However, this is
accomplished through cell signaling or communication
(tissue interaction) which plays a vital part in the
differentiation and growth various cells found in the
embryo. This mechanism allows the complex multi
cellular organism to function and develop in a very
coordinated manner. Cellular communication during
the developmental process should occur between cells
or group of cells at the right time and intensity for
normal development. Cells that influence another are
called inducing cells while the affected one is called
the induced cell. With signaling, various changes
(Figure 6.4: A to D) may occur in the induced cells.

Figure 6.4. Mechanisms of development in

the cells of the embryo.

For cell differentiation and specialization, a very informative video entitled “How cells
become specialized?” may help you further understand the concept. Another video also shows
cell specialization and stem cells. See the links below:

1. https://youtu.be/t3g26p9Mh_k
2. https://youtu.be/gwAz_BtVuLA
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EMBRYONIC DUPLICATION AND TWINNING

The term “twins” means two individuals which develop in the same pregnancy in animals
that are normally monotocous (animals that produce one individual per pregnancy). There are 2
distinct types of twins, monozygotic and dizygotic. Dizygotic twins arise from 2 ova, derived
from 2 separate ovarian follicles, each fertilized by separate spermatozoa during a single
breeding cycle. Monozygotic twins arise from a single ovum fertilized by a single spermatozoon.
There are different stages in early embryological development when monozygotic twins may
arise (see Figure 6.5).

Multiple births can be caused by any of the following causes:

1. Fertilization of separately ovulated gametes of the female (common)
2. Complete or partial separation of cleavage-stage blastomeres and blastocysts
3. Duplication during gastrulation.

Embryos from multiple births are

classified as free (unattached to each
other) or conjoined, and based on the
uniformity in size they are further
classified as symmetrical or
asymmetrical.
1. Free (separate), symmetrical
dizygotic twins
This classification includes
most normal twins (fraternal)
where they develop from separate
zygotes thus are called dizygotic
twins. Each embryo develops
independently with its own
separate set of fetal membranes
although in some species, fusion
of extraembryonic membranes
occurs. In the cow, this includes
fusion of the allantoic blood
vessels, which allows blood to be
exchanged between fetuses. If the
twins are of different sexes, the
exchange results in the abnormal
development of the genital system
of the female and the production of
a free martin.
2. Free symmetrical,
monozygotic twins
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These are identical twins

derived initially from a single Figure 6.5. Stages in embryological development at which
zygote, which separates or monozygotic twins may arise: (A) Formation of 2 blastocysts within a
duplicates. The two blastomeres zona pellucida; (B) Formation of 2 inner cell masses within a single
produced form the first cleavage
blastocyst; (C) Division of a blastocyst as it emerges from the zona
may separate and each form an
pellucida; and (D) Formation of 2 primitive streaks which arise from a
embryo. This separation more
single embryonic disk.
frequently occurs later during the
cleavage or blastocyst stages. Two separate sets of extraembryonic membranes will be
formed whenever twinning is initiated during these stages.
An abnormally small rupture in the zona pellucida permits some but not all of the
blastocyst to break free. This may allow the formation of monozygotic twin embryos provided
that both blastocysts contain part of the embryonic disk (each half develop normally). In
some instances, the embryonic disk will split into two separate parts immediately prior to
gastrulation wherein there is usually some sharing of the extraembryonic membranes
between the two fetuses.
3. Free asymmetrical twins
This type of twins originates from monozygotic or dizygotic twins. In free asymmetrical
twins, one embryo is normal while the other is rudimentary in development and survives by
being attached to the blood supply of the fetal membranes of the normal twin. The abnormal
twin (also called holocardius, acardiac fetus, amorphous globosus, anidian (formless)
fetus) has no recognizable body form and consists of skin with pigment and hair, connective
tissue, bone, teeth, muscle and rudimentary digestive organs. Specific craniofacial
structures can sometimes be identified. The abnormal twin is usually surrounded by its own
amnion. Among domestic animals acardiac twins are common in cattle.
This abnormal twinning should be differentiated from the condition known as mummified
fetus or lithopedion (stone child) wherein the development of the normal twin is arrested.
Rather than being reabsorbed, the twin becomes dehydrated and shrunken. Mummified
fetus can cause dystocia (abnormal labor or parturition) or, if retained leads to uterine
infection or interfere with a subsequent pregnancy. Mummified fetuses are most commonly
found in cows.
4. Conjoined or fused symmetrical twins
These twins are monozygotic in origin and represent incomplete division of one embryo
into two components, usually at some time during the primitive streak stage. If the twins are
nearly complete, they are generally called diplopagus (2-fold joined), the preferred
equivalent of siamese twins. Conjoined twins are usually cl
assified by the point at which they are joined (Gr. pagos - "that which is fixed.").
A. Conjunction never involving the heart or umbilicus
1. Craniopagus - cranial union only.
2. Pygopagus - posterior union of the rump.
B. Conjunctions always involving the umbilicus (midline conjunctions)
1. Thoracopagus - anterior union of the upper half of the trunk, twins are joined at the
sternal region of the thorax, facing each other. It is the most common form of
conjoined twins, it always involves sharing the heart.
2. Cephalopagus - anterior union of the upper half of the body with two faces on
opposite sides of a conjoined head. Extremely rare. The heart is sometimes involved.
A combination of types 1 and 2 is called epholothoracopagus.
3. Parapagus (sometimes called diprosopus) - lateral union of the lower half, extending
variable distances upward. Heart sometimes involved.
4. Ischiopagus: Anterior union of the lower half of the body. Heart not involved.
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5. Omphalopagus/Abdominopagus: joined at the abdomen, often with partially fused

intestines.

Duplication of one part of the fetus' axial (and adjacent) structures is also possible. This
usually arise during the elongation or regression of the primitive streak. This set of anomalies is
described by using the prefix di- (or tri-, tetra-, etc.) and the appropriate region-specific suffix
(e.g. dicephalus= two heads, diprosopus= two heads, dicaudatus= two tails, tetrabrachius= two
pairs of thoracic limbs, tetrascelus= two pairs of pelvic limbs, etc.)

Figure 6.6. The eight types of

conjoined twins: (1) cephalopagus,
(2) thoracopagus, (3)
omphalopagus, (4) ischiopagus, (5)
parapagus, (6) craniopagus, (7)
pygopagus, (8) rachipagus.

Source:https://www.researchgate.net/
publication/

271830992_Monochorionic_Twin_Pregnancy-_Potential_Risks_and_Perinatal_Outcomes

5. Conjoined asymmetrical twins

These twins are unequal in size (heteropagus) and consist of one reasonably normal
individual (autosite) and an extra body part (parasite - smaller, less formed twin dependent
upon the other) attached to it. A frequent manifestation of this is the formation of an extra limb
attached at the back of the animal, this is called notomelus (notos = back, melus=limb).
Another example of asymmetrical twinning is a normal animal with an extra set of pelvic limbs
projecting caudally from its ischial arch. Asymmetrical conjoined twins usually arise after
gastrulation when specific organ forming regions called fields (eg: limb filed, heart field, eye
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field, etc.) are beginning to organize. Another one is Fetus in fetu: Situation wherein an
imperfect fetus is contained completely within the body of its sibling.

Early Nourishment
The embryo is nourished with embryotroph which is composed of:
1. Histotrophe – aka. Uterine milk; responsible for early nourishment while the placenta is not
yet well established; absorbed by imbibition of trophoblast cells
2. Hemotrophe – nutritive material from maternal blood; absorbed through the allantochorion or
vitellochorion

Short videos about monozygotic and dizygotic twins maybe of help for you to visualize
how these phenomena occur:
1. https://youtu.be/E_-dHExmWuU
2. https://youtu.be/_2g-Aaing3Y

Learning Activity:

Let’s have Critical Thinking questions for this module. Answer the questions below:

1. Discuss how the therapeutic use of embryonic stem cells be advantageous or

disadvantageous.
2. Explain the use of artificial twinning in domestic animals like cattle, in order to
increase reproductive efficiency.

Deadline for submission of answers to this module: November 15, 2021.