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The early development of Chick: Cleavage & gastrulation.

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 The early development of chick: cleavage and gastrulation

Zoology
Lesson: The early development of chick: cleavage and
gastrulation
Lesson Developer: Dr Meena Yadav
College/Department: Maitreyi College, University of
Delhi

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 The early development of chick: cleavage and gastrulation

TABLE OF CONTENTS

 Introduction
 Cleavage in chick
 Gastrulation
 The hypoblast
 The primitive streak
 Formation of endoderm and mesoderm
 Regression of primitive streak
 Cell movement in the ectoderm
 Avian Embryo: Axis Formation
 pH and dorsal-ventral axis
 Gravity and anterior-posterior axis
 Left-right axis
 Gene expression on the right side of primitive
streak
 Gene expression on the left side of primitive streak
 Summary
 Exercise/practice
 Glossary
 References
 Suggested readings
 Useful web links

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 The early development of chick: cleavage and gastrulation

INTRODUCTION
There have been many animal models for studying embryology. Aristotle was the first
scientist to follow the early development of the chick till 3-weeks. Since then, chick has
become a favorite organism for embryological studies. Advantages of chick as the
model organism for such studies are:
1. It is accessible all around the year and is easy to breed.
2. Its developmental stage can be accurately predicted at any given temperature.
This makes it possible to collect large number of embryos at same stage of
development
3. The embryos can be surgically manipulated
4. Since the stages of its development are similar to mammals, it provides a
suitable medium to study various stages of development and also provides an
insight into the development of mammalian embryos.

CLEAVAGE IN CHICK
In birds, fertilization occurs in the oviduct of the female just before the albumin and
shell are laid upon it. The chick egg is telolecithal i.e. the cytoplasm has been pushed to
one corner of the egg and the rest of the egg is filled with yolk. A small disk of the
cytoplasm is present on top of a mound of yolk (Figure 1).

Figure 1: The chick egg

Source: Author

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 The early development of chick: cleavage and gastrulation

The cleavage in eggs of the birds is discoidal meroblastic. The cleavage divisions occur
only in the cytoplasm present at the animal pole in the form of a blastodisc, its
diameter being only 2-3 mm. The first cleavage occurs at the center of the blastodisc
and other cleavage divisions follow soon, to create a single layer of cells called
blastoderm (Figure 2). The cleavage divisions do not extend into the yolk, so the
peripheral cells remain in contact with yolk and the rest of the cells are also connected
with each other.

Figure 2: Cleavage in chick egg

Source: Author

Later the vertical and equatorial divisions continue in the blastoderm and produce a
sheet of tissue consisting of six to seven layers. These cells are now connected with
each other by tight junctions. As the divisions continue, the blastodermal cells absorb
fluid from the albumin and secrete it such that the fluid accumulates between the cells
and between the cells and yolk. The space between the yolk and blastoderm is called
subgerminal cavity. The cells present in the deep center of the blastoderm die and
are shed, thus creating a one-cell thick area pellucida. This part forms the actual
embryo while the peripheral ring of the blastoderm does not shed the cells and form the
area opaca (Figure 3).
Marginal zone or marginal belt is the name given to a thin layer of cells between the
area pellucida and area opaca.

GASTRULATION

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 The early development of chick: cleavage and gastrulation

Gastrulation is an early event during embryogenesis in vertebrates where the group of

cells, formed after cleavage divisions arrange themselves into three layers i.e.
ectoderm, mesoderm and endoderm. These three layers are precursors of future
organs in the vertebrate body. However, the events of gastrulation differ slightly in all
vertebrates. Here, we will discuss events that occur during avian gastrulation.

The hypoblast
In hen, at the time of laying eggs, the blastoderm contains as many as 20,000 cells.
Most of the cells of the area pellucida remain at the surface of the animal pole and form
the epiblast, while the rest of the cells of the area pellucida delaminate themselves
and migrate into the subgerminal cavity, where they form polyinvagination islands
or primary hypoblast. These cells exist in clusters of 5-20 cells each. The posterior
margin of the blastoderm contain a local thickening of cells called as Koller’s sickle.
As soon as the delamination of the hypoblast cells starts, a sheet of the cells from the
Koller’s sickle region starts migrating to the anterior side and join the polyinvagination
islands, thus forming secondary hypoblast (Figure 3).

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 The early development of chick: cleavage and gastrulation

Figure 3: Migration of cells to form two layers i.e. epiblast and

hypoblast with blastocoel separating them
Source: Author

VALUE ADDITION: Things to know

Heading: Cell movements of primitive streak of the chick embryo
Body Text:

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 The early development of chick: cleavage and gastrulation

Cell movements of the primitive streak of the chick embryo. (A-C) Dorsal view of the
formation and elongation of the primitive streak. The blastoderm is seen at (A) 3–4 hours,
(B) 7–8 hours, and (C) 15–16 hours after fertilization. The early movements of the
migrating epiblast cells are shown by arrows. (D-F) Formation of notochord and
mesodermal somites as the primitive streak regresses, shown at (F) 19–22 hours, (E)

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 The early development of chick: cleavage and gastrulation

23–24 hours, and (F) the four-somite stage. Fate maps of the chick epiblast are shown for
two stages, the definitive primitive streak stage (C) and neurulation (F). The endoderm
has already ingressed beneath the epiblast, and convergent extension is seen in the
midline.
Source: http://www.devbio.biology.gatech.edu/?page_id=52 CC

The blastoderm now has two layers i.e. epiblast and hypoblast, separated by a cavity
called blastocoel. The entire avian embryo is formed from the cells derived from the
epiblast. The hypoblast cells form the portions of the extraembryonic membranes like
yolk sac and stalk, which links the yolk mass to endodermal digestive tube. The epiblast
cells, thus, form the embryo and some extraembryonic membranes.

VALUE ADDITION: Things to know

Heading: Signal for initiation of primitive streak formation
Body Text:

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Source: http://www.mun.ca/biology/desmid/brian/BIOL3530/DEVO_04/ch04f11.jpg

The primitive streak

The presence of a primitive streak is characteristic feature of the avian, reptilian and
mammalian gastrulation. The primitive streak appears as a thickening of the epiblast at
the posterior side near the Kolliker’s sickle. This thickening is the result of the
ingression of endodermal precursor cells from epiblast into the blastocoel and also the
migration of cells from the lateral regions of the epiblast towards the center. As the
migration of these cells continues towards the primitive streak, the streak elongates

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 The early development of chick: cleavage and gastrulation

towards the anterior side or future head.

Simultaneously, the secondary hypoblast cells continue to migrate towards the anterior
side. The primitive streak extends to 60-75% of the length of area pellucida.
The primitive streak is important in establishing the axes of the embryo. It lies length
wise in an anterior-posterior axis; the cells migrate from its dorsal side to the ventral
side and it also separates left side from the right side. The cells that are closer to the
primitive streak form the medial (central) structures while cells away from it form the
distal (lateral) structures.

Figure 4: Migration of cells towards the primitive streak and

formation of primitive groove
Source: Author
The cells keep on coming towards the primitive streak. As they pile up, some of them
start converging at the center of the streak from both sides. This causes formation of a
groove at the center of the streak called as primitive groove (Figure 4). This groove
then serves as the opening through which the migrating cells pass into the blastocoel.
Thus, the primitive groove is similar in function to the amphibian blastopore.

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VALUE ADDITION: Things to know

Heading: Migration of cells during chick gastrulation
Body Text:

Source:
http://s723.photobucket.com/user/alphamalik/media/47_13ChickGastrulation.jpg.html

The anterior end of the primitive streak contains a thickening of the cells known as
primitive knot or Hensen’s node. This node contains a funnel shaped depression at
the center, called primitive pit, which acts as a passage for the cells to pass to the
blastocoel (Figure 5). The Hensen’s node is analogous to dorsal lip of the blastopore in
amphibians.
The above figure shows that once the primitive streak is formed, its cells start
migrating through the primitive groove and primitive pit into the blastocoel. These cells
migrate anteriorly and form the foregut, head mesoderm and notochord. Another
population of cells passes through the lateral portions of the primitive streak and
gives rise to most of the endodermal and mesodermal derivatives.

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 The early development of chick: cleavage and gastrulation

Figure 5: Side view of the primitive streak stage during chick

embryogenesis
Source: Author
The cells that enter the inside of the avian embryo ingress as individuals after they have
undergone epithelial-mesenchymal transformation. The cells at the Hensen’s node and
at the entire length of the primitive streak are broken from the basal lamina
(delamination) and are released into the embryo. This is thought to be assisted by a
190-kDa protein called scatter factor. This protein is secreted by the cells themselves
as they enter the primitive streak. The epithelial cells are converted into the
mesenchymal cells in several ways by the scatter factor. It is thought to downregulate
the expression of E-cadherin and thus, prevents its function as well.

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VALUE ADDITION: Things to know

Heading: Formation of primitive streak and endodermal layer in chick
Body Text:

Source: https://www.ucl.ac.uk/cdb/research/stern/stern_lab/Fig3_copy.jpg
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VALUE ADDITION: Things to know

Heading: Function of E-cadherin
Body Text: E-cadherin is a cell-adhesion molecule and helps in attachment of
adjacent cells with the help of catenins and actins.

Source: http://www.clinsci.org/cs/118/0451/cs1180451a02.gif

Formation of endoderm and mesoderm

The cells start migrating towards the primitive streak and eventually enter the
blastocoel.
(1) The first cells to start migration form the pharyngeal endoderm of the foregut.
Once they enter the blastocoel, they migrate anteriorly and displace the
hypoblast cells, such that the hypoblast cells become confined to a region in the
anterior portion of the area pellucida. This specific region is called germinal
crescent, and contains the precursors of the germ cells. These precursor cells
then migrate to the gonads through the blood vessels.
(2) The next group of cells entering the blastocoel through the Hensen’s node, also
move anteriorly, but not as far as those of the pharyngeal endoderm. They
establish themselves between the endoderm and the epiblast to form
prechordal plate and head mesenchyme. All these cells move anteriorly and

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 The early development of chick: cleavage and gastrulation

push up the anterior midline region of the epiblast to form the head process. This
places the head of the embryo anterior (rostral) to the Hensen’s node.
(3) The next cells in line become the chordamesoderm (notochord) cells. These
cells extend anteriorly to the presumptive midbrain and meet the prechordal
plate. The chordamesoderm extends from some anterior portion of Hensen’s
node till some posterior portion of it. The chordamesoderm at the level of the
Hensen’s node and caudal to it form the hindbrain and trunk respectively.
(4) Simultaneously, the cells continue to migrate from the lateral sides into the
primitive streak. As they enter the blastocoel through the primitive groove, they
separate into two layers:
(i) The deep layer joins the hypoblast in the middle and displaces the hypoblast
cells to the sides. These cells give rise to the endodermal organs of the embryo
and most of the extraembryonic membranes. The hypoblast cells give rise to rest
of the extraembryonic membranes.
(ii) The second layer spreads itself between the endoderm and the epiblast and
forms a loose layer of the cells. These middle layer cells form the mesoderm and
thus the mesodermal derivatives and some extraembryonic membranes.
After 22 hours of incubation, most of the presumptive endodermal cells lie at the
interior of the embryo while the presumptive mesodermal cells continue their
migration inwards.

Regression of the primitive streak

While the mesodermal cells continue their migration, the primitive streak starts to
retreat such that the Hensen’s node moves from the center of area pellucida to a
posterior position. Thus, it leaves the dorsal axis of the embryo and the notochord. The
notochord is laid down starting from the future midbrain towards the posterior side as
the Hensen’s node retreats.
The formation of the anterior and rest of the notochord is contributed by different set of
cells: the anterior part of the notochord is formed by the cells that entered through
the Hensen’s node. The posterior notochord (after somite 17 in chick) is formed from
the mesodermal tissue, which was formed from the cells ingressed through the lateral
sides of the primitive streak. The posterior portion of the notochord extends till the tail
of the embryo. The regression of the primitive streak continues till the Hensen’s node
reaches the posterior most portion and forms the anal region.

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VALUE ADDITION: Things to know

Heading: Prospective cardiac cell migration to form heart tube in chick
Body Text:
“In avian embryos, the prospective cardiogenic cells are located in the epiblast and primitive streak
(Figure). Within the epiblast, these cells are bilaterally distributed on both sides of the primitive
streak, caudal to the node. A few hours later (PS5), the rostral half of the primitive streak, with the
exception of the node, contains the prospective cardiogenic cells (Figure 1), and also the cells that are
going to form the endoderm which underlies the pre-cardiac mesodermal cells at both sides of the
embryo. Progeny of cells from this region will contribute to all layers of the heart tube, including
endocardium, myocardium, and parietal pericardium.
Later (HH4) in gastrulation (Figure 1), pre-cardiac cells from the epiblast invaginate through the
primitive streak, and move bilaterally and cranially, to form the bilateral cardiogenic mesoderm,
located in the anterior position, and constituting the primary cardiac field, also called the
heart-forming region. At this moment, the pre-cardiac mesoderm is surrounded by the adjacent
endoderm, which comes from the more cranial part of the primitive streak, and which plays a crucial
role during cardiac specification.
From stage HH7 begins the organization of the heart rudiment pre-cardiac splachnic mesoderm,
which will form both of the primitive endocardial tubes, surrounded by the underlying endoderm.
From stage HH9, both primitive endocardial tubes fuse in the mid-line to form the primitive heart
tube, structurally organized in concentric layers of endocardium and myocardium.”

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Fig. Schematic diagrams showing the pre-cardiac cells, from stage HH3 (PS3) to stage
HH10, in the developing chick embryo. Red indicates the mesodermal pre-cardiac cells, and green
the endodermal cells that are related to cardiogenesis. The section shows the migration of epiblast
cells ingressing through the primitive streak at stage HH4.
Source: Carmen López-Sánchez, Virginio García-Martínez. Molecular determinants of cardiac
specification Cardiovascular Research185-195.DOI: http://dx.doi.org/10.1093/cvr/cvr127
http://cardiovascres.oxfordjournals.org/content/cardiovascres/91/2/185/F1.large.jpg

At this point of development, all the presumptive endodermal cells and the mesodermal
cells lie inside the embryo and the epiblast represents the presumptive
ectodermal cells. As the cells migrate and take their respective future positions in the
embryo like head, notochord etc., the anterior-posterior axis is established in the
embryo. The cells at the anterior portion of the embryo are ahead in their development
and start forming the organs even as the cells at the posterior portion are still
undergoing gastrulation. Thus, there is an obvious spatial and temporal difference in
the stages during the chick embryogenesis. For the next few days, the anterior portion
runs ahead with the development as compared to the posterior portion.

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VALUE ADDITION: Things to know

Heading: Regression of the primitive streak in chick
Body Text:

Source: http://www.mun.ca/biology/desmid/brian/BIOL3530/DEVO_04/ch04f10.jpg

Cell movement in the ectoderm (epiboly= 4 days)

At the time when the endodermal and mesodermal precursor cells are migrating to
their prospective positions in the embryo, the ectodermal precursor cells start
proliferating. These cells start surrounding the yolk by their migration called epiboly.
This process takes about 4 days to complete.
The precursor ectodermal cells migrate underneath the vitelline envelope, however,
only the cells of the outer edge of area opaca attach to the vitelline envelope. These
cells are unique as they can extend numerous large cytoplasmic processes (500 µm)
into the vitelline envelope. These processes act as the locomotor apparatus (called as
filopodia) of these marginal cells and help them in pulling other ectodermal cells
around the yolk.
The filopodia bind to a protein present in the vitelline envelope called fibronectin. This
binding helps in the migration of ectodermal cells beneath the vitelline envelope. If
experimentally, the contact between the marginal cells and fibronectin is broken, the

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 The early development of chick: cleavage and gastrulation

epidermal migration stops as filopodia retract.

At the end of gastrulation in chick, the three germ layers take their respective positions,
as in other vertebrates:
i) The ectoderm surrounds the yolk and forms the outermost covering of the
embryo.
ii) The endoderm replaces the hypoblast
iii) The mesoderm establishes itself between these two layers.

Avian embryo: Axis formation

The signals for the axis formation in chick embryo come from the start of the
embryogenesis i.e. cleavage. The various factors affecting the formation of avian axis
are:
pH and dorsal-ventral axis
The first axis that is formed is the dorsal-ventral axis, which differentiates the back and
belly side of the chick in the embryo. It is critical for the formation of the first new layer
of cells i.e. hypoblast. This axis is established due to the cleaving blastodermal cells.
These cells create a pH gradient between the albumin above the blastodisc (pH= 9.5,
basic) and subgerminal space below it (pH=6.5, acidic) (Figure 6).
The blastodermal cells transport water and Na+ ions from the albumin into the
subgerminal space, thus creating a membrane potential difference of 25 mV across the
epiblast cell layer. The membrane potential is positive at the ventral side of the epiblast.
This exchange of ions separates the two sides of the epiblast:
i) One side having negative membrane potential and facing basic albumin. This
side becomes the future dorsal side
ii) The other side having positive membrane potential and facing the acidic
subgerminal space fliud. It becomes the future ventral side of the embryo.
If the pH of the two sides is reversed or the membrane potential is reversed, the axis
formation is also reversed experimentally.

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 The early development of chick: cleavage and gastrulation

Figure 6: pH and dorsal-ventral axis formation

Source: Author
Gravity and anterior-posterior axis
The gravity plays an important role in converting radially symmetrical blastoderm into
bilaterally symmetrical embryo. When the ovum is passing through the reproductive
tract of the hen, it is rotated for 20 hours in the shell gland. The rate of the spinning
is 10-12 revolutions per hour. This spinning causes the rearrangement of the yolk
components such that its lighter components lie beneath one side of the blastoderm.
Due to this, that portion of the egg tips up and is the place from where the formation of
the primitive streak initiates. This part then becomes the posterior portion of the
embryo.
The molecular signals that make this part of the egg as posterior portion and initiate
gastrulation are not fully understood. Any part of the marginal zone can initiate
formation of the primitive streak. It has been proved by experiments (Spratt and
Haas, 1960) in which the blastoderm is separated into many parts, each having its
own marginal zone. Each part was able to form its own primitive streak.
Once the posterior marginal zone (PMZ) is formed in the marginal zone, it prevents
the other regions of the marginal zone to initiate formation of any other primitive streak
and also allows the gastrulation to commence. Thus, it appears that PMZ may contain
cells analogous to those of Nieuwkoop center of the amphibians, which give signals for
the initiation of gastrulation.
If a part of the posterior marginal zone tissue (not including Koller’s sickle) is grafted at
the anterior region, it induces the formation of the primitive streak and Hensen’s node.
At this region, the TGF-β signaling and β-catenin localization in the nucleus, coincide.
The posterior region of the embryo is the only source of Vg1.
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Important points to remember:

S.No. Important points of chick gastrulation

1 The Hensen’s node is formed by the cells from the epiblast and the middle
layer cells of the anterior Koller’s sickle.
2 Posterior portion of the Koller’s sickle form the posterior portion of the
primitive streak
3 Dorsal mesoderm: forms the central nervous system in the ectoderm
overlying it.
4 The cells of the Hensen’s node and its derivatives: they secrete
Chordin, Noggin and Nodal proteins. They inhibit the action of the BMPs and
dorsalize the ectoderm and mesoderm.

The avian Hensen’s node is analogous to the amphibian dorsal blastopore lip
because:
i) It is the site where the gastrulation begins
ii) It is the region whose cells become chordamesoderm
iii) It is the region whose cells can establish new embryonic axis when transplanted
into new locations of the gastrula.
iv) If the Koller’s sickle is transplanted into a new embryo, it causes formation of
new axes
v) The middle layer cells of the Koller’s sickle express goosecoid as do the cells of
Spemann organizer in amphibians.
The neural induction in chick is different from that of fishes or amphibians. Unlike in
amphibians, in chick embryos, ectopic expression of chordin in non-neural epiblast
cells do not cause neural induction and BMPs do not inhibit neural induction.
The important factor that causes positive regulation of neural induction in chicks are
Fibroblast Growth Factors (FGFs). They are produced in Hensen’s node and
primitive streak and generate neuronal phenotypes in the epiblast. The signals
controlling the anterior neuron production are still unknown but studies by Diaz and
Schoenwolf (1990) and Darnell et al (1999) suggest that anterior visceral
endoderm may provide such signals.

Left-right axis

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The vertebrate body is distinctly divided into left and right axis, and so are its internal
organs located on either of the sides. For example, heart and spleen generally lie on
the left side of the body while the liver occupies the right side. There are two major
proteins that regulate the left and right axis formation:
i) Nodal (a paracrine factor)
ii) Pitx2 (a transcription factor)
The mechanisms by which these two genes i.e. nodal and pitx2 regulate the left-right
axis formation varies in different groups of vertebrates.
Some of the common genes expressed during the lefty-right axis formation are
(Figure 7):
 Gene expression on the right side of the primitive streak:
a) Sonic hedge-hog: the expression of this gene occurs on both the sides of the
primitive streak. However, once the primitive streak reaches its maximum length,
transcription of its mRNA stops on the right side of the embryo. This is due to
presence of a protein activin and its receptor, which actually downregulate the
expression of shh gene.
But at the same time activin signaling activates the expression of fgf-8. FGF-8
prevents the expression of the caronte gene. In the absence of caronte gene, the
BMPs are able to block the expression of two genes i.e. nodal and lefty-2.
b) Snail gene (cSnR): The suppression of the nodal and lefty-2 genes activates
the snail gene, expressed exclusively on the right side of the embryonic organs.

 Gene expression on left side of the primitive streak:

On the left side of the body, the lefty-1 gene is expressed and it blocks the expression
of fgf-8, while shh activates the caronte gene. The protein Caronte, is a paracrine factor
that inhibits BMPs so that nodal and lefty-2 genes remain activated. Also, due to
downregulation of BMPs, the lefty-1 is also expressed properly on the ventral midline
structures. Nodal and Lefty-2 activate pitx2 but suppress snail. The protein, Lefty-1 in
the ventral midline prevents the Caronte signal from passing to the right side of the
embryo.
Pitx2 plays an important role in creating asymmetry in the embryo as has been seen in
Xenopus. If we experimentally induce the expression of nodal or pitx2 on the right side
of the embryo, the symmetry of the embryo is reversed or randomized on the left or
right side. If any of the steps of the left-right axis formation are blocked or interfered,
the symmetry is randomized.

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Figure 7: Different genes involved in the left-right axis formation

in chick
Source: Author
VALUE ADDITION: Things to know
Heading: Cross-section of the chick embryo at primitive streak stage
Body Text:

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Source:
http://www.mun.ca/biology/desmid/brian/BIOL3530/DB_03/fig3_17.jpg

Some results of the experiments during chick gastrulation:

 If follistatin, a blocker of activin is added, the asymmetry conferred by shh

disappears and the position of the heart tube is randomized.
 If beads soaked in activin are placed on the left side of the Hensen’s node, the

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expression of the sonic hedgehog gene is suppressed. This blocks the expression of
the nodal. Under such circumstances, the heart may be present on either left or
the right side.
 If certain cell are grafted on the right side of the Hensen’s node, which secrete
sonic hedgehog, the above condition results.
In all these cases, nodal is induced symmetrically in the lateral plate of mesoderm and
the heart has equal chances of being present on either of the sides.
Once the three germ layers have occupied their respective positions in the embryo, the
organogenesis can begin. Although, several genes or factors that play role during
gastrulation and in the axes formation have been identified and their role ascertained,
there still lie several lacunae in the in-depth understanding of the exact mechanisms of
events occurring during avian gastrulation.

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SUMMARY
1. The chick egg is filled with huge amount of yolk and little cytoplasm at the point
of sperm entry.
2. The cleavage divisions start at the blastodisc and after subsequent divisions
form a single layer sheet of cells called blastoderm
3. The cleavage divisions form two distinct regions in the embryo i.e. area opaca
and area pellucida. The region between them is known as marginal zone.
4. The formation of the primitive streak and gastrulation starts at the posterior
marginal zone.
5. The primitive streak is formed from the deep cells of the posterior marginal zone
and anterior epiblast cells.
6. The primitive streak moves anteriorly and simultaneously Hensen’s node is also
formed
7. The cells that migrate through the Hensen’s node into the blastocoel form the
chordamesoderm (notochord). These cells extend upto the presumptive
midbrain where they meet the prechordal plate.
8. The prechordal induces the formation of the forebrain; chordamesoderm forms
the midbrain, hindbrain and spinal cord.
9. Cells that enter the blastocoel from the lateral side of the primitive groove form
the endoderm and in the process they displace the hypoblast.
10. The mesodermal cells follow the endodermal cells into the blastocoel.
11. The ectoderm, present on the surface, surround the entire yolk by epiboly
12. The dorsal-ventral axis in birds is dependent on the pH difference on two sides
of the embryo while gravity determines the anterior-posterior axis.
13. The left-right axis is dependent on the expression of the nodal gene on the
left-side of the embryo. Nodal gene activates the pitx2 gene which leads to
formation of the structures on the left side of the body.

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 The early development of chick: cleavage and gastrulation

EXERCISE/PRACTICE
Q.1. Give contribution of the following scientists:

i) Spratt and Haas

ii) Diaz and Schoenwolf
Q.2. Define
i) Hansen’s node ii) Primitive groove
iii) Primitive streak iv) Koller’s sickle
Q.3. Fill in the blanks:
i) The formation of ______________________________ in the marginal zone
prevents the initiation of formation of primitive streak.
ii) The gene whose expression is exclusively upregulated on the right side of the
embryo is _______________________________.
iii) FGF-8 prevents the expression of ___________________ gene.
iv) _____________________________ blocks the action of activin.
v) The central nervous system in the chick is formed by the
________________________________________________.
vi) _______________________ in chick is known as the equivalent of amphibian
dorsal blastopore lip.
vii) The ovum spins a t a rate of ________________________ while moving
through the hen’s reproductive tract.
viii) The membrane potential across the epiblast cell layers is
______________________________________________.
ix) The cells of the area opaca produce cytoplasmic processes known as
______________________ into the vitelline envelope.
x) The hypoblast cells contribute to the formation of
________________________________________________.

Q.4. Draw a comparison between avian Hensen’s node and amphibian dorsal lip of the
blastopore.
Q.5. List the genes involved in the development of structures on the left and right side
of the chick embryo.

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 The early development of chick: cleavage and gastrulation

Q.6. Label the parts in the following figures:

(a)

(b)

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The early development of chick: cleavage and gastrulation

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 The early development of chick: cleavage and gastrulation

Q,6. Write short notes on the following:

i) PMZ
ii) Polyinvagination islands
iii) Epiboly of the ectoderm
iv) Formation of the Hensen’s node
v) Role of pH in dorsal-ventral axis formation
vi) Regression of primitive streak
Q.7. Discuss the influence of gravity on the anterior-posterior axis formation.
Q.8. True/False
i. Cleavage in avian eggs is holoblastic
ii. Nodal and Pitx2 are involved in left-right axis formation in chick
iii. Blastoderm is a 5-7 layer thick layer of cells
iv. The space between yolk and blastoderm is called subgerminal cavity
v. E-cadherin helps in cell signaling in chick embryo
vi. Marginal zone lies between area opaca and area pellucida
vii. Scatter factor helps in delamination of cells from primitive streak
viii.Koller’s sickle lies on the anterior side of the chick embryo
ix. Blastocoel is lined by epiblast and hypoblast
x. The secondary hypoblast includes cells from epiblast and Koller’s sickle

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 The early development of chick: cleavage and gastrulation

GLOSSARY
Area opaca: The cells at the peripheral ring of the blastoderm are not shed and form
area opaca.
Area pellucida: The deep cells of the blastoderm die and are shed, giving rise to single
layer of cells called area pellucida. These cells form the actual embryo
Blastocoel: The cavity between epiblast layer and hypoblast layer
Blastoderm: A single layer of cells formed after cleavage divisions in chick at the
animal pole.
Blastodisc: The name given to small amount of cytoplasm in the avian eggs.
Epiblast: The cells of the area pellucida are known as epiblast cells
Filopodia: The cytoplasmic processes of the precursor ectodermal cells at the outer
edge of area opaca, which maintains contact with vitelline envelope. They act as the
locomotor apparatus and help pulling other ectodermal cells around the yolk during
epiboly.
Germinal crescent: The specific region at the anterior portion where the hypoblast
cells get accumulated and form the precursors of the germ cels
Hensen’s node/primitive knot: Thickening of the cells at the anterior portion of the
primitive streak
Koller’s sickle: Local thickening of the cells at the posterior margin of the blastoderm
Marginal zone/marginal belt: The thin layer of cells between area pellucida and
area opaca.
Polyinvagination islands/ Primary hypoblast: The cells delaminating from the
epiblast and exist in the groups of 5-20 each
Posterior marginal zone (PMZ): The cells that initiate gastrulation and start the
formation of the primitive streak, thereby inhibiting other cells of the marginal zone to
initiate other primitive streaks.
Primitive groove: The groove formed at the centre of the primitive streak due to
ingression of cells
Primitive pit: A funnel shaped depression at the centre of the Hensen’s node
Primitive streak: Thickening of the cells at the middle line of the blastoderm in
anterior-posterior direction, starting from the posterior side near Koller’s sickle
Scatter factor: A protein (190-kDa) secreted by the cells themselves as they enter the
primitive streak and is required for their delamination from it
Subgerminal cavity: The space between yolk and blastoderm

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 The early development of chick: cleavage and gastrulation

Telolecithal eggs: The eggs which contain large amount of yolk and the small amount
of cytoplasm lies at one end.

REFERENCES

1. Developmental Biology: S.F. Gilbert 7th Ed.

 SUGGESTED READINGS
1. An introduction to embryology: B.I. Balinsky and B.C. Fabian

 USEFUL WEB LINKS

1. http://www.ncbi.nlm.nih.gov/books/
2. https://embryology.med.unsw.edu.au/embryology/index.php/Book_-_The_Ea
rly_Embryology_of_the_Chick_5
3. http://www.devbio.biology.gatech.edu/?page_id=52

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