VETERINARY DEVELOPMENTAL ANATOMY

MODULE 4

EARLY STAGES OF DEVELOPMENT IN BIRDS AND MAMMALS:

CLEAVAGE, GASTRULATION AND NEURULATION

Learning Objectives:

After the completion of the module, you will be able to:

1. classify the different types of cleavage;
2. explain the process of cleavage, gastrulation and neurulation in mammals and avians

CLEAVAGE

Right after fertilization the zygote undergoes cell division, or cleavage. Thus, one cell divides into
two; the daughter cells, called blastomeres, then cleave into four; these cleave into eight, and so on.
During this early stage of development, the zygotes of different animal species vary in appearance. This
difference is mainly attributed to the variation in the yolk content and distribution of the yolk in the
cytoplasm of the female egg. The following are the classifications of egg based on the amount and
distribution of yolk:

Classification based on amount Classification based on Species

of yolk distribution of yolk

Micro/oligolecithal- small Isolecithal- equal distribution Mammals

amount

Mesolecithal- moderate amount Centrolecithal- yolk in the Most amphibians

middle of the egg

Macro/megalecithal- large Telolecithal- concentrated at Fish, birds, reptiles

amount one pole

Based on the abundance and distribution of the yolk, cleavage can be identified as either
holoblastic or meroblastic. Holoblastic cleavage occurs when the entire ovum divides and the
blastomeres produced are of either equal or unequal size. In megalecithal ova, mitosis is restricted to the
animal pole where the cytoplasm is devoid of yolk. This type of division is referred to as meroblastic
cleavage. In fish, birds and reptiles, because the site of cleavage is confined to a disc-shaped area or
the animal pole, this type is also known as discoidal cleavage. See image below:
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Holoblastic vs. Meroblastic Cleavage

Zygote of animals having telolecithal type of egg undergo meroblastic (incomplete) type of
cleavage, wherein the daughter cells are not completely separate from each other thus, in its very early
stage forms a syncitium (a cell with many nuclei). On the other hand, those with isolecithal eggs
undergo holoblastic (complete) division, yielding to completely separate daughter cells or blastomeres.
In avians, cleavage first occurs at the center of the central white area of the ovum called the
blastodisk, the exact location of the nucleus and cytoplasm of the zygote. This area is surrounded by a
darker peripheral region blending with the yolk, the periblast. At its early stage of development, the
very young chicken embryo is a syncitium (first four to five cell cleavage). Later on, mononucleated
blastomeres are formed, they proliferate rapidly at the center. These cells separate from the underlying
yolk, creating a space called the subgerminal cavity. The blastomeres overlying this area are called
central blastomeres, wherein the body of the embryo will be formed. The more peripherally located
blastomeres are called the marginal blastomeres, these will form the extraembryonic membranes of
the avian embryo.
Stages of cleavage in the avian zygote from first
cleavage to formation of blastoderm
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Source: McGeady TA, Quinn PJ, FitzPatrick ES and Ryan MT.2006. Veterinary
Embryology. UK: Blackwell Publishing.

In mammals, mitotic divisions occur in right angles up to the third mitotic division, this is then
followed by random division of the blastomeres. When the embryo consists of a hundred or more cells it
forms a compact solid mass, called a blastula or morula (from its resemblance to a mulberry). At this
stage the blastomeres lose their spherical shape and become tightly attached to each other. Prior to this
stage the corona radiata is shed while the zona pellucida stays intact.
In most species the compact mass of cells then transforms itself into a single layer of cells
forming a hollow sphere, the blastocyst. This transformation is accomplished by the secretion of fluid by
the blastomeres which are collected at the center forming a central cavity called the blastocoel.

During the blastula stage the zona pellucida ruptures, this allows the embryo to break free,
enormously enlarge and alter its shape. Some examples of these changes are: dog – blastocyst becomes
pear shaped; horse - remains spherical; cattle, pig and sheep embryos elongate tremendously (at the
rate of 1cm/hour).

In animals with an isolecithal zygote, the blastocyst cells are divided into two groups:
1. outer blastomeres- become flattened and form tight junctions resulting in reduced
permeability to fluids. They develop the capacity to secrete fluid they are destined to become
trophoblasts (equivalent of the marginal blastomeres in birds) which form the fetal chorion
& amnion (fetal membranes)
2. inner blastomeres- form gap junctions to maximize intercellular communication. They are
destined to become inner cell mass or epiblast which forms the body of the embryo, yolk sac
and allantois.

* In mammals the trophoblast cells covering the inner cell mass degenerate or move peripherally
to expose the embryonic disk of the blastocyst, thus called the blastodisk. In most primates, the
overlying trophoblast cells remain thus the embryonic disk remains to be located beneath the sheet of
trophoblast thus termed as inner cell mass.

Development from the first cleavage to the blastocyst stage of mammals

Source: Fletcher TF and Weber AF. Veterinary Developmental Anatomy: Veterinary Embryology Class Notes

GASTRULATION

Following the blastula stage is gastrulation, the stage of embryonic development wherein the
three primary germ layers are formed through complex cell migrations. The three primary germ layers
are the 1. Ectoderm - outermost wall, 2. Mesoderm - middle layer and 3. Endoderm - innermost wall.
This stage of development is considered to be the most critical since in this period the potentials
of specific tissues to develop begin to be limited or restricted.
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Like in the cleavage stage, the events that happen during gastrulation varies between species
especially between placental (mammals, primates, etc.) and nonplacental animals (like birds).

Gastrulation in Avians

Gastrulation in birds involves the central blastomeres which constitutes the area pellucida (clear
area). This region is termed as such due to its lighter appearance, which is attributed to the presence of
the subgerminal cavity that separates the central blastomeres from the underlying yolk. However, the
area occupied by the marginal blastomeres appear darker due to their attachment to the yolk, thus called
the area opaca (opaque area).

Before gastrulation occurs, four important events

during the blastula stage must take place:
1. Early blastula stage - cells of varying sizes mingle
with each other
2. Segregation - the large central blastomeres
containing more yolk segregate to form the roof of
the subgerminal cavity, other large blastomeres
gather at the caudal end of the embryonic disk.
3. Delamination - the large blastomeres detach from
the roof of the subgerminal cavity and migrate
ventrally to from a new layer of cells, the hypoblast.
4. Formation of the hypoblast - a new layer of cells
is established and completely divides subgerminal
cavity into two. The larger space above the hypoblast
is now the blastocoel while the smaller cavity below
is still called the subgerminal cavity.

Formation of hypoblast and primitive streak

in avian embryo

As soon as the hypoblast layer is formed,

gastrulation then starts. Major events in avian
gastrulation are the following:
1. Radial expansion of the epiblast
and formation of the primitive
streak - epiblast cells at the cranial
region expand towards the caudal end
and aggregate in the caudal midline.
This aggregation extends cranially to
give rise to a thickened median area
called the primitive streak. The
primitive streak will serve as the
landmark for the future longitudinal
axis of the embryo. This area will
continue to elongate up to the extent
of the entire blastodisk. The primitive
streak resembles a hairpin when
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viewed dorsally. It presents a slightly enlarged cranial tip called the primitive node or
Hensen's node, which has a deep hole in the middle, called the primitive pit.
2. Formation of the mesoderm and endoderm - A median depression appears on the
primitive streak, the primitive groove. This is formed due to the movement (involution) of the
epiblast cells from the midline of the primitive streak towards the blastocoel. The first cells that
migrate invade the hypoblast layer, displacing the original hypoblast towards the periphery. The
epiblast cells that invaded the hypoblast becomes the intraembryonic endoderm, the
displaced hypoblast becomes the extraembryonic endoderm. The other epiblast cells that
leave the center of the primitive streak migrate peripherally and form a loose mesenchymal
population of cells located between the epiblast and the endoderm, this shall form the
mesoderm.
3. Regression of the primitive streak and formation of the notochord - this is the
last stage in gastrulation in birds. The cranial two third of the epiblast cease to expand while the
cells in the primitive streak continue to migrate and add to the mesenchymal population. With the
continued cell migration and with no cells to replace lost cells in the primitive streak, regression
occurs. Simultaneous with the regression of the primitive streak is the deposition of epiblast cells
in the midline beneath the original epiblast (becomes the ectoderm), forming the notochord.
Gastrulation in birds is completed when the primitive streak has completely regressed

Gastrulation in birds starts at approximately 24-30 hours after fertilization (about 6 hours after
laying). The primitive streak reaches its maximum length (around 2mm.) at about 18-19 hours of
incubation, it completely regresses by 60 hours (2.5 days) of incubation.

Gastrulation in Mammals and Primates

Gastrulation in these species is very much similar with that in birds although with some distinct
differences due to the necessity of forming the extraembryonic membranes and eventually the placental
membranes that will establish contact and mediate metabolic interaction with the mother.
During the blastula stage the epiblast cells delaminate from the inner surface of the embryonic
disk and migrates beneath the trophoblast to form a thin sheet of cells lining the interior of the
blastocyst, this is the hypoblast. The cavity enclosed by the hypoblast becomes the primitive gut
(archenteron). Most of the hypoblast (endoderm) will remain outside the embryo to form the yolk sac,
while a small portion of the tube shall remain inside the embryo to give rise to the gut tube.
The primitive streak like in birds is formed at the epiblast of the embryonic disk which is relatively
a small area compared to the extraembryonic trophoblast. Some of the epiblast cells will migrate beneath
the primitive streak to give rise to the intraembryonic endoderm, intraembryonic mesoderm, lateral
mesoderm, somites and notochord.
In animals where the blastocysts are remarkably elongated (pig, sheep and cattle), the lateral
mesoderm quickly expands away from the embryonic disk while simultaneously splitting into two layers.
The two layers originating from the lateral mesoderm are the:
1. Somatic mesoderm - layer closely next to the overlying ectoderm of the embryonic disk
and trophoblast and
2. Splanchnic (visceral) mesoderm - layer closely adjacent to the underlying endoderm and
hypoblast. The somatic mesoderm together with the embryonic ectoderm and trophoblast
will form the somatopleure while the splanchnic mesoderm, endoderm and hypoblast will
form the splanchnopleure.
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Gastrulation in pig embryo (Day 13-15 of development)

1: Trophectoderm; 2: Epiblast; 3: Primitive streak; 4: Mesendodermal precursor cells; 5: Intra-embryonic
mesoderm; 6: Chorioamniotic fold; 7: Chorion; 8: Extra-embryonic mesoderm; 9: Endoderm; 10: Hypoblast; 11:
Coelom; 12: Surface ectoderm; 13: Mesoderm; 14: Neural groove; 15: Notochord; 16: Amniotic cavity.

Source: Hyttel P, Sinowatz F and Vejslsted M. 2010. Essentials of Domestic Animal Embryology. Edinburgh:
Saunders Elsevier.

NEURULATION

This is the stage of embryonic development that follow after gastrulation, this stage is
characterized by the first development of the neural plate and subsequently, the neural tube. At the same
time the early development of the heart, gut and somites take place during this stage.

Early Development of the Central Nervous System

1. Formation of the neural plate - formed by

thickening of the surface ectoderm along the dorsal
midline of the embryonic disk
2. Development of the neural groove and neural
folds - the neural groove is formed by the depression
of the neural plate at the midline, this is bound on
both sides by elevations called neural folds. The
neural folds are thickest and highest at the anterior
part of the embryo, this is due to the rapid mitotic
activity in this region.
3. Formation of the neural tube - the neural folds
gradually elevate and converge at the dorsal midline
of the embryonic disk. The fusion starts at the
posterior brain region and continues cranially and
caudally (like a zipper). The formed neural tube
eventually separates from the overlying surface
ectoderm to further develop and give rise to the
central nervous system.
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Neurulation process

Source: Fletcher TF and Weber AF. Veterinary

Developmental Anatomy: Veterinary Embryology Class
Notes

During the formation of the neural tube, the cephalic region of the embryo rapidly develops and begins to
elevate above the surface of the embryonic disk. Together with this elevation of is its rostral elongation to form
the head process. This head process together with the cephalic neural tube will over grow the underlying
ectoderm and endoderm resulting to the formation of the subcephalic pocket, a horizontal furrow or space
beneath the head process.
Together with growth of the head process, the lateral body folds are formed. The downward folding of
the ectoderm lateral to the neural tube forms these folds which are continuous with the subcephalic pocket.
During the course of development these furrows become deeper causing the separation of the head and
eventually more parts of the body from the underlying extraembryonic tissues. Subsequently, the same
structure will be formed at the caudal end of the embryo, this will progress cranially to meet the earlier formed
lateral body folds at the level of the umbilicus.

Development of the neural tube and the neural crest.

The lateral edges of the neural plate are elevated and become the neural folds. The depressed midregion of the
neural plate is called the neural groove. The neural folds continue to rise, appose in the midline and fuse to create
the neural tube, which becomes covered by future epidermal ectoderm. As the neural folds rise and fuse, cells at
the lateral border of the neuroectoderm (neural crest cells) begin to dissociate from their neighbors, undergo an
epithelio-mesenchymal transition, and leave the neuroectoderm. 1: Surface ectoderm; 2: Neural plate; 3: Neural
groove; 4: Neural crest; 5: Neural tube; 6: Spinal ganglion; 7: Anterior neuropore; 8: Posterior neuropore; 9:
Notochord; 10: Primitive node; 11: Primitive streak; 12: Somites.

Source: Hyttel P, Sinowatz F and Vejslsted M. 2010. Essentials of Domestic Animal Embryology. Edinburgh:
Saunders Elsevier. Pp. 96.
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Other developments during neurulation involving the other systems are presented below, they
will be discussed in detail later in the development of the different body systems.
Early development of the heart
1. Formation of the endocardial primordium by the cardiogenic mesoderm
2. Development of the endocardial tube
3. Fusion of the two endocardial tubes

Early development of the gut tube

1. Formation of the foregut due to the separation of the head process form the extraembryonic
membranes
2. Formation of the hindgut due to the formation of the tail fold at the caudal tip of the embryo

LEARNING ACTIVITY:
1. Watch The following videos and animations on the links below to further understand and
visualize the lesson:
A. Early Embryogenesis: Cleavage
https://www.youtube.com/watch?v=9JLQDmrj7fI&feature=share
B. Gastrulation in Birds
Gastrulation in chick(L=1)-most comprehensive note
C. Gastrulation in Mammals
https://www.youtube.com/watch?v=ADlYn0ImTNg
D. Neurulation
https://www.youtube.com/watch?v=lGLexQR9xGs
2. Select one stage in early embryogenesis that interests you most (cleavage, gastrulation or
neurulation). Using any materials that you could see in your home, make a video clip
demonstrating the stage of development that you have selected. Be as creative as you can in
your demonstration. Videos should be a maximum of 5 minutes only. Upload your videos in the
respective file folder in our official Google Drive.