Unit 15 Development of Chick

UNIT 15

DEVELOPMENT OF CHICK

Structure
15.1 Introduction Fully Formed Gastrula

Objectives 15.6 Neurulation in Chick

15.2 Structure of Egg of Chick Mechanisms of Neural Plate
Formation
15.3 Fertilisation
Morphogenesis of Mesodermal
15.4 Cleavage and Blastulation
Derivatives
15.5 Gastrulation
15.7 Folding of Embryo
Role of Hypoblast
15.8 Development of Extra-
Fate Map Embryonic Membranes
The Gastrulation Process: Development of Amnion and
Formation of Primitive Streak Chorion
Completion of Endoderm Development of Allantois
Regression of Primitive Streak 15.9 Hatching
Epiboly of Ectoderm 15.10 Summary
Characteristic Features of 15.11 Terminal Questions
Avian Gastrulation
15.12 Answers
Comparison with Amphibian
Gastrulation

15.1 INTRODUCTION
Different animals have evolved a variety of strategies of development.
However, since all animals are related, the basic mechanism of early
development has been conserved in the course of evolution, and so there are
some important similarities in early embryonic development of all metazoan
animals as you have already learnt in Block 3.

This unit speaks about development of chick as an example of an amniote

organism. Recall that amniotes are those vertebrates (reptiles, birds and
mammals) that have a water sac or amnion surrounding the developing
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organism protecting it from the external environment. Chick has been one of
the first model organisms to be studied in detail as it is easy to maintain and
large enough to be manipulated surgically and genetically during all stages of
development.

You will study about strictly coordinated sequential changes that take place
during the course of chick development viz. fertilisation, cleavage, formation of
morula, blastula, gastrula and organogenesis that give rise to the new
individual.

Chick egg is macrolecithal which undergoes discoidal and meroblastic

cleavage. Cleavage results in morula which is converted into blastula. During
gastrulation there is displacement of the parts of blastoderm so that the
presumptive endodermal and mesodermal cells are removed and brought into
the interior of embryo through morphogenetic movements. Neurulation, the
next step, consists of the formation of first the neural plate and then the neural
tube. Then due to future changes and development of extra embryonic
membranes, the new organism emerges.

Objectives
After studying this Unit you should be able to:

describe the structure and fertilization of chick egg;

describe cleavage and formation of blastula in chick egg;

discuss the process and mechanism of chick gastrulation;

describe the morphogenetic processes that lead to the formation of

neural tube and neural plate in chick;

discuss the development of extra embryonic membranes in chick; and

compare the development of chick with that of frog.

15.2 STRUCTURE OF EGG OF CHICK

The egg is about 3.0 cm in diameter and is macrolecithal. It is entirely filled up
by the yolk and over it, i.e., in animal pole lies a small cytoplasmic disc with a
nucleus. This disc is called blastodisc.

The yolk contains 48.7% water, 32.6% phospholipids and fats, 16% proteins,
1% carbohydrates and 1.1% other chemical molecules.

Yolk is covered by a layer of dense viscous albumen which forms a thin

chalaziferous layer around the vitelline membrane of the yolk. This dense
albumen forms two twisted cords or chalazae, one at each end of the egg.
They are formed by rotation of the egg during its movement through the
oviduct (Fig.15.1).
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Unit 15 Development of Chick

Fig.15.1: Diagrammatic longitudinal section of a hen’s egg.

Around the chalaziferous layer is a thick layer of watery albumen. All albumen
is secreted by the upper glandular walls or magnum of the oviduct. The
functions of albumen are to provide nutrition to the embryo, serves as a water
store and also acts as protective envelope for the embryo protecting it from
mechanical and chemical injuries.

The albumen and yolk also contain a variety of enzymes, vitamins, pigments
and phosphorus. The isthmus part of the oviduct secretes two shell
membranes made of tough keratin fibres matted together.

The two shell membranes are closely applied except at the blunt end of the
egg where they are separated by an air space formed after the egg is laid. The
nidamental glands of the oviduct secrete a porous, calcareous shell over the
two membranes which soon hardens after the egg is laid. The shell is pierced
by a large number (about 7,000) of fine pores filled with a protein related to
collagen. The diameter of the pores varies from 0.04 to 0.05 mm. These pores
allow exchange of gases (oxygen and carbon dioxide) during respiration of the
developing embryo. Once the egg is laid there is a cooling in the egg
temperature and a contraction of the egg mass creating an air space between
the two shell membranes at the broad end of the egg.

SAQ 1
Choose the correct word from the parentheses.

a) The egg of hen is (macrolecithal/microlecithal).

b) Cytoplasmic disc with a nucleus at animal pole is called

(blastoderm/blastodisc).

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15.3 FERTILISATION
The ova are released from the ovary in the form of primary oocytes. The
second maturation division occurs after ovulation in the oviduct. The released
primary oocyte in the body coelom is grasped and swallowed by the ostium of
the oviduct.

The secondary oocyte or ovum after second maturation division is surrounded

by several (5 or 6) sperms which enter in it (polyspermy), but only one sperm
succeeds in the fertilization process. The nucleus of one sperm fuses with the
female nucleus (amphimixis) and the nuclei of other sperms degenerate.

As the fertilised egg passes downward inside the oviduct, it rotates, undergoes
cleavage and various accessory egg membranes are laid down over the
developing egg. Thus, fertilisation is internal. Various egg membranes
secreted around egg are albumen, shell membranes and shell. The egg is laid
24 hours after fertilisation, and its further development takes place, when the
egg is incubated by the female. Incubation must continue steadily for 21 days
at a temperature of 37.44°C.

15.4 CLEAVAGE AND BLASTULATION

As you know, cleavage, depends to a large extent upon the amount,
distribution and orientation of yolk in the egg. As hen’s egg is macrolecithal, so
the cleavage in chick is discoidal meroblastic. That is, cleavage is confined
only to the germinal disc or blastodisc situated at the animal pole (Fig.15.2).
Cleavage starts about three hours after fertilization while the egg is still in the
hen’s oviduct. First two cleavages are at right angles to each other in the
centre of blastodisc. These cleavage furrows do not cut the germinal disc
completely through in the vertical plane.

The third set of cleavage furrows is vertical, cutting across the second set of
vertical furrows. The fourth cleavage furrow is also vertical and circular cutting
across all the cleavage furrows, forming eight central blastomeres which are
surrounded by eight marginal blastomeres. Thus, these cleavage furrows
separate the daughter central blastomeres from each other, but not from the
yolk. The central blastomeres are continuous with the underlying yolk at their
lower ends (Fig.15.3). At this time the maternal determinants are directing the
cleavage, but by the 7thor 8th cleavge division (when there are 128 cells), the
switch from maternal to zygotic gene expression occurs.

Fig.15.2: Surface view of egg showing the blastodisc with primitive streak. A
166 denotes anterior and P denotes posterior part.
Unit 15 Development of Chick

Fig. 15.3: Surface view of germinal disc of hen’s egg showing early cleavage. A)
Two-cell stage; B) Six-cell stage; C) Twenty-cell stage; D) Late
cleavage stage E) Showing relationship between yolk and surface
blastomeres.

The marginal blastomeres are continuous with the uncleaved cytoplasm at

their outer edges. Further cleavages are irregular. The central cells divide
more rapidly. The marginal cells also divide by the appearance of new
horizontal and radial furrows. The newly formed inner cells of marginal
blastomeres are added to the central cells, resulting in the increase of volume
of this area. The radial furrows extend peripherally and these peripheral cells
are still continuous with the uncleaved peripheral cytoplasm (Fig.15.3).

In later stage of cleavage, the blastomeres of the central area become

separated from the underlying yolk due to the appearance of a horizontal
cleavage in these cells. This cleavage extends peripherally cutting the inner
ends of the blastomeres. Thus, a space also appears beneath the central cells
which also extends peripherally as the horizontal cleavage extends outward.

The cavity beneath the central cells, i.e., in between central cells and yolk is
called the subgerminal cavity, which is filled with a fluid diffused from the
albumen through vitelline membrane. Thus, due to further cleavage the
blastodisc becomes cellular, called the blastoderm- a round disc, 5 to 6 cells
deep in the centre but only 1 to 2 cells deep at the periphery.

The appearance of subgerminal cavity separates the blastoderm from the

underlying yolk. Some of the cell layers from the center of blastoderm are
shed and die so that only a single or a few layer of cells in this area remains
making the area translucent. Thus the central area is known as area
pellucida, but the marginal cells remain overlapping the yolk and appear
opaque and darker. Therefore, this area is termed area opaca. These cells in
area opaca process the yolk but do not contribute in embryo formation (see
Fig.15.4A). Between the area pellucida and area opaca is a thin layer of cells
which is known as marginal zone. The embryo is now called the blastula.. At
blastula stage the embryo reaches the uterus. It may be compared to the
blastula of Amphioxus and frog, its sub-geminal cavity is equivalent to the
blastocoel, the blastoderm is the animal pole, and the yolk is the vegetal pole. 167
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The egg is laid by the female about the time the blastula is formed or even a
little later. At the time of laying, the blastoderm is composed of 20,000 to
60,000cells. Most of the cells of the area pellucida remain at the surface,
forming an “upper layer” called the epiblast (Fig.15.4A).

Shortly before the egg is laid some cells delaminate from the epiblast and
ingress into the subgerminal cavity in clusters forming the primary hypoblast
(Fig.15.4B). Shortly after the egg is laid, a small thickening of the epiblast,
called Koller’s sickle, is formed at edge of the area pellucida which is now
marked as the posterior end of the embryo. In between the area opaca and
Koller’s sickle is a belt like region of cells called the posterior marginal zone
(PMZ). At the same time a sheet of cells begins to delaminate and migrates
from the posterior edge of the area pellucida under the surface. These
delaminated cells at the posterior edge of area pellucida gradually link up with
each other and with the primary hypoblast, to form a continuous layer of
flattened cells, which lie over the yolk on the floor of the subgerminal cavity.
This layer is called as secondary hypoblast or endoblast and the upper
layer of the blastoderm is the epiblast containing ectoderm and mesoderm
cells. Hypoblast is exclusively composed of endoderm cells (Fig.15.4C).

Fig.15.4: Formation of subgerminal cavity, epiblast and hypoblast and Koller’s

sickle in the chick blastula.
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Unit 15 Development of Chick
SAQ 2
a) Which type of cleavage is observed in chick?
b) What is blastoderm?

15.5 GASTRULATION
In chick, gastrulation is a prolonged process. It is initiated soon after the
beginning of incubation after laying of the egg and is completed in about 4
days. The actual process of gastrulation is preceded by some pre-gastrular
movements of certain cells resulting in their separation from the blastoderm
and formation of a lower layer called the hypoblast. Most of the cells,
however, remain in the upper layers of the blastoderm which now constitute
the epiblast (Fig. 15.6).

15.5.1 Role of Hypoblast

In the earlier section you have already learnt how the hypoblast forms from
the blastoderm.The hypoblast thus formed expands and spreads peripherally
to give rise to a complete thin layer below the remaining part of blastoderm,
now called the epiblast. The hypoblast and epiblast are joined together at the
margin of area opaca and the space between them is the blastocoel. The
structure of the embryo is now somewhat similar to that of the frog blastula
but the hypoblast is not the precursor of either ectoderm, mesoderm or
endoderm. It is later displaced by endodermal cells derived from epiblast.

Hypoblast contributes cells for parts of some extra-embryonic membranes

especially the yolk sac and the stalk linking the yolk mass to the endodermal
digestive tube but none at all for the formation of the body of the embryo.
However, the hypoblast has an important role in the development of the
embryo. Its removal at an early stage stops all further development until a new
hypoblast is regenerated from the epiblast. Hypoblast induces the formation of
the primary embryonic axis (the primitive streak) in the epiblast and
determines its orientation. Hypoblast cells also provide chemical signals that
specify the migration of epiblast cells. However, the three germ layers of the
embryo proper (plus the amnion, chorion, and allantois i.e. extraembryonic
membranes) are formed solely from the epiblast.

15.5.2 Fate Maps

A chart or diagram showing the prospective fate of each part of blastula or
embryo at any stage of development is called a fate map. Fate maps of the
blastula of chick have been prepared artificially by using the vital stains such as
carmine or carbon (charcoal) particles or radioactive thymidine. Fate map
shows that blastomeres of area opaca do not form any part of the embryo
proper, they form only extraembryonic membranes.

The epiblast and hypoblast of area pellucida have different fates in the course
of embryonic development. The fate maps prepared by the use of tritiated
thymidine have shown the following structures. In the centre of area pellucida 169
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lies a small area destined to produce the notochord. Posterior to it, in the
median plane is found an elongated oval area of presumptive endoderm which
will form the gut (Fig. 15.5).

Fig. 15.5: Fate map of chick blastoderm immediately prior to gastrulation. The
primitive streak is not yet formed but it will extend eventually to the
region of the notochord. .

Further toward the posterior edge of area pellucida lies the extraembryonic
endoderm which will form the lining of yolk sac. To the right and left of the
presumptive notochord and endoderm, and posterior to extra-embryonic
endoderm lie the various subdivisions of presumptive mesoderm, i.e.,
prechordal plate or head mesoderm, mesodermal somites, lateral plate
mesoderm and extra-embryonic mesoderm.

As a result of morphogenetic movements during gastrulation the cells from the

various areas reach their respective specific destinations forming the ectoderm
on the surface, endoderm below and mesoderm between the two.

The anterior half of epiblast is the presumptive ectoderm containing central

presumptive neural plate area, anterior to it is the presumptive embryonic
epidermis and outer to it in the form of complete ring is the extraembryonic
ectoderm.

15.5.3 The Gastrulation Process: Formation of

Primitive Streak
Gastrulation in all amniotes including eutherian mammals is related to a
characteristic structure called the primitive streak formed on the epiblast
surface during the first 10-18 hours of incubation at 37.5-385°C. The primitive
streak can be considered as the equivalent of an elongated blastopore lip of
amphibian embryos. It forms as a result of convergence of epiblast cells to the
dorsal midline of the blastoderm. Dyemarking experiments and time-lapse
photography indicate that the primitive streak first arises from Koller’s sickle
and the epiblast above it the beginning of primitive streak formation is first
170 indicated by a thickening in the posterior marginal zone region of area
Unit 15 Development of Chick
pellucida immediately after the formation of endoblast. The thickening narrows
and elongates growing anteriorly in the centre of area pellucida. When fully
formed the primitive streak is a narrow structure with a groove (primitive
groove) in its floor along its length flanked by a fold (or ridge) on either side. If
we consider the primitive streak is homologous to the blastopore lip then the
primitive groove is homologous to the amphibian blastopore, The primitive
streak extends anteriorly upon about three fourth the length of area pellucida
where it ends in a deep pit called Hensen's Node with thick borders (Fig.
15.6).The center of Hensen’s node contains a funnel-shaped depression
through which cells can enter the embryo to form the notochord and
prechordal plate. Hensen’s node is the functional equivalent of the dorsal lip of
the amphibian blastopore or the organizer.

Fig. 15.6: Initiation of primitive streak formation from the thickening known as
Koller’s sickle. The streak corresponds to the blastopore lip of the
amphibians. 171
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During the formation of primitive streak the shape of area pellucida gradually
changes from circular to pea shaped with the broad side anterior and narrow
side posterior. This change is due to convergence of cells toward dorsal
midline beginning at posterior end and progressing anteriorly but stopping
where the Hensen's Node is formed. Primitive streak marks the median
anterior-posterior axis of the embryo and establishes bilateral symmetry
(Fig.15.7 and. see Box 15.1). It also establishes the dorso- ventral axis of the
embryo.

Fig. 15.7: Migration of endodermal and mesodermal cells through the primitive
streak. A) Diagram of transverse section through a 17 hours embryo,
illustrating the blastocoel; B) Diagram of medial section through the
same embryo, showing that those cells migrating through Hensen’s
Node condense to form the notochord (head process); C) Stereogram
of gastrulating chick embryo, showing the relationship of the primitive
streak, the migrating cells, and the two original layers (epiblast and
172 hypoblast) of the blastoderm.
Unit 15 Development of Chick
Box 15.1: Setting up the antero-posterior axis in chick embryo.

How do we know what will be the posterior side in the blastoderm where
the primitive streak will start to develop? As early as 1828, von Baer gave a
general rule that enabled the antero- posterior axis to be predicted in the
majority of eggs. The rule states that ‘if the egg is horizontal with the
pointed end to the right then the tail of the embryo should be towards the
observer’. This happens because the egg undergoes a continuous rotation
when it is in the uterus and this is usually in the same direction relative to
the sharp and blunt ends of the egg. The embryo and yolk do not rotate
along with the outer surface of the egg but are nevertheless tilted in the
direction of rotation .It is thought that the rotation of the egg causes the
lighter components of the yolk to accumulate under one side of the
blastoderm. These yolk components are probably maternal determinants
for development. This tilted end of the blastoderm becomes the posterior
marginal zone where the primitive streak is initiated. And the opposite end
becomes the anterior side towards which the primitive streak will extend.
The cells of PMZ initiate gastrulation and once PMZ is formed it controls
the surrounding regions of the margin between area pellucida and area
opaca and prevents the formation of any other primitive streak. Thus we
see that a radially symmetrical cleavage pattern gives rise to a bilaterally
symmetrical organism and all because of gravity and rotation of the egg!

Movements of Epiblast Cells

The cells for the presumptive mesodermal and endodermal organ areas pass
from the epiblast into the blastocoel below through the Hensen's Node or
primitive groove of the primitive streak. This occurs by the cells sheets
converging toward the Hensen's Node or the streak. On reaching there the
cells change shape becoming ''bottle cells", the sheet breaks up into separate
cells which pass into the blastocoel individually by involution through the
Hensen's Node or the primitive streak. Once inside the blastocoel, the cells
flatten and continue to migrate as streams of loosely connected mesenchyme
vertically down or laterally and anteriorly.

As cells enter the primitive streak, they undergo an epithelial-to-mesenchymal

transformation and the basal lamina beneath them breaks down. In contrast to
the amphibian gastrulation where sheets of cells involute during gastrulation,
avian grastrulation is an ingression of epiblast cell which form a
loose mesenchyme within the blastocoel. The first cells to enter blastocoel
are those of the presumptive foregut endoderm located immediately around
the Hensen's Node through which they move in. On entering the blastocoel
they move anteriorly and ventrally and displace the cells of the anterior part of
hypoblast. Later, infolding of this endodermal layer forms the foregut.

Next, the cells of presumptive chorda-mesoderm migrate into the blastocoel

also through the node and then move anteriorly in the mid line just below the
overlying epiblast to form the head mesoderm and anterior part of notochord
called the head process. Later, the remaining endodermal and mesodermal
cells of the epiblast migrate through the anterior and posterior regions of the
streak, respectively. Once inside the blastocoel they form two streams of 173
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migrating cells. One stream contains the endodermal cells which move down
along the midline pushing the hypoblast to the side and forming a continuous
sheet with that of the foregut endoderm. The other stream contains cells of
presumptive somite and lateral plate mesoderm. They remain within the
blastocoel, move laterally and anteriorly to take up positions on either side of
the forming notochord as a loose sheet of mesoderm between the epiblast and
hypoblast. The movement of cells within the blastocoel is facilitated by
hyaluronic acid secreted by the ectodermal cells of epiblast. This
polysaccharide accumulates in the blastocoel and coats the surface of the
incoming cells which keeps them separate allowing their migration as
individual cells.

SAQ 3
Choose the appropriate word given in the parenthesis.

i) Most of the cells remain in the upper layers of the blastoderm which
constitute the (epiblast/hypoblast).

ii) As a result of morphogenetic movements during gastrulation the cells

from various areas reach their respective specific destinations forming
the (ectoderm/endoderm) on the surface.

iii) During the formation of primitive streak, the shape of area

(opaca/pellucida) gradually changes from circular to pea shaped with
broad side anterior and narrow side posterior.

iv) The cells for the presumptive mesodermal and endodermal organ areas
pass from the epiblast into the (blastocoel/gastrocoel) through the
Hensen’s Node or primitive groove of the primitive streak.

v) The (polysaccharide/disaccharide) accumulates in the blastocoel and

coats the surface of incoming cells.

15.5.4 Completion of Endoderm

The first cells that migrate through the anterior part of streak form the
endoderm. By 22 hours of incubation most endodermal cells have migrated
internally while the mesodermal cells continue to migrate till later. As the
mesodermal ingression continues, the primitive streak begins to regress
moving the Hensen’s node from near the center of area pellucida towards a
posterior position. As the Hensen’s node recedes backward, and the
notochordal process elongates, the presumptive endoderm of the middle and
posterior part of the gut, located just behind the node, migrate inside as an
endodermal strip beneath the notochord. These will give rise to the
endodermal organs, major part of the gut of the embryo and most of the
extraembryonic membranes. The remaining part of extraembryonic
membranes will be formed by the hypoblast and cells of the area opaca.

174 In chick no archenteron is formed during gastrulation (Figs.15.8 and 15.9).

Unit 15 Development of Chick

Fig.15.8: Primitive streak and germ layer formation in the chick embryo. A-
Surface view of the epiblast; In B, C and D the left side pictures show
the surface and the right side depict the pattern of the mesodermal
areas as they spread over the hypoblast below; E- is a diagrammatic
cross section showing the involution of mesoderm through the
primitive streak.

Fig.15.9: Ingression of endodermal and mesodermal cells through the Hensen’s

node and the primitive groove. The migrating cells of the endoderm
from the Hensen’s node move anteriorly to form the head process and
the migrating endoderm cells move deep laterally over the hypoblast.
The mesodermal cells form a loosesheet of mesenchyme which will
form the mesodermal organs. 175
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15.5.5 Regression of Primitive Streak

As the inward migration of cells through the primitive streak ends progressively
along its anterior to posterior regions, the streak gradually regresses in the
same direction. As the streak shortens, the Hensen’s node, which forms its
anterior end, also moves in a posterior direction (Fig.15.9). Simultaneously,
the presumptive notochordal cells still remaining in the epiblast migrate inward
through the posteriorly moving node adding to the length of the notochord. The
regressing streak thus leaves behind in its path the dorsal axis of the embryo
represented by the posteriorly lengthening notochord (Fig.15.9). By the time
the streak disappears and the Hensen’s node arrives at the final position at the
posterior border of area pellucida all the presumptive endodermal and
mesodermal cells have left the epiblast which now consists of only the
ectodermal cells. Regression of streak begins after about 22 hours of
incubation and is completed in the next about 20 hours. It should be noted that
unlike amphibian gastrulation, differentiation of axial structures and some
organs such as neural tube, somites, foregut, heart, occurs cephalo-caudally
along with the progress of later stages of gastrulation in chick embryos
(Fig.15.10). This means that head structures are already forming while
migration of cells is still going on in the regressing primitive streak (See
Fig.15.11A and B).

Fig. 15.10: Regression of the primitive streak, leaving notochord in its wake.
Time represents hours after achieving maximum length by the
primitive streak.

Fig. 15.11: Dorsal view of regression of primitive streak. Note that while the
streak is regressing from anterior to posterior region the organs are
176 already forming in the anterior region.
Unit 15 Development of Chick
15.5.6 Epiboly of Ectoderm

As the presumptive endodermal, notochordal and mesodermal cells migrate

inward the epiblast is made entirely of ectodermal cells (of the embryonic
epidermal and neural ectoderm). Simultaneously the ectodermal precursors
proliferate, expand as a sheet beneath the vitelline membrane outward from
the area opaca ultimately encircling the yolk through epiboly (recall
morphogenetic movents from Unit 12). The marginal cells of area opaca are
different from other blastodermal cells, they can extend enormous cytoplasmic
extensions or fillopodia to form a firm contact with the inner surface of
vitelline membrane.. It appears that these marginal cells of area opaca migrate
outward along the inner surface of the vitelline membrane with the help of
these pseudopodia and at the same time drag along the expanding sheet of
ectodermal cells of which they are the leading members. It is believed that the
fillopodia connect to protein fibronectin in the vitelline membrane and if the
fibronectin is broken experimentally, ectoderm migration stops. Similarly
removal of the vitelline membrane inhibits epibolic movements and expansion
of ectoderm.

15.5.7 Characteristic Features of Avian Gastrulation

i) Formation of hypoblast and its important role in formation of the axis and
orientation of embryo.

ii) Presence of the cells of all the three germinal areas in the epiblast.

iii) Formation of the primitive streak and its regression in later stages of
gastrulation.

iv) Absence of archenteron formation. Gastrular cell movements include

ingression, involution, convergence, divergence and epiboly.

v) Cephalo-caudal differentiation of axial structures and growth of the

embryo during gastrulation.

15.5.8 Comparison with Amphibian Gastrulation

On comparison of chick gastrulation with amphibian gastrulation you will find

many similarities along with important differences. Primitive streak is
analogous to the blastopore although there is no actual pore. The Hensen’s
node represents the dorsal lip of amphibians and the folds on the lateral sides
of primitive groove represent the lateral lips. However, there is no ventral lip.
As in amphibians, the prechordal endoderm and chorda mesoderm of chick
migrate inward through the Hensen’s node (dorsal lip) and the remaining
endoderm and mesoderm do so by involution over the lateral folds of the
streak (lateral lips). The topographic locations of the presumptive ectodermal
chordamesodermal; mesodermal and endodermal area relative to each other
in the epiblast are nearly the same as on the surface of amphibian blastula. As
in amphibians, the involuting mesodermal and endodermal cells change shape
to become bottle cells in the gastrulating chick embryos as well. 177
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15.5.9 Fully Formed Gastrula

Gastrula is fully formed when primitive streak completely disappears. The fully
formed gastrula consists of three germ layers-ectoderm, chordamesoderm and
endoderm. The ectoderm and chorda-mesoderm remain in continuity along
the axis of primitive streak. The endoderm is also united with the mesoderm
and ectoderm at the anterior and posterior end of streak (Fig.15.12).

Fig.15.12: Cross section of an early chick embryo, illustrating the relationship

of the germ layers.

SAQ 4
i) List the types of morphogenetic movements of cells that occur during
gastrulation in chick embryos.

ii) What is the role of hypoblast in chick embryos?

iii) Cells of which presumptive germinal layers are constituents of the

epiblast in chick embryo?

iv) How do mesodermal and endodermal cells migrate inward?

v) Which structures are formed from cells of area opaca?

vi) List similarities between gastrulation in chick and amphibians.

15.6 NEURULATION IN CHICK

You already know from Unit 13 when studying about the development of frog
that the vertebrate brain and spinal cord start their development as a flat plate
of neuroepithelial cells that folds up along most of its length to form a tube.
The process of forming this neural tube is called neurulation. 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
neurulation has begun in the anterior part of the embryo, the posterior region
is still in the process of gastrulation. At a time when the neural folds are just
about to form in the posterior region, in the anterior region the neural folds
178 have already started fusing forming the neural tube (Fig.15.13).
Unit 15 Development of Chick

Fig.15.13: Dorsal view of chick embryo of 25-26 hours with 5 pairs of somites.
Neural folds approaching each other fuse and form neural tube in the
cephalic region; but in the posterior region gastrulation is still
occurring and primitive streak and neural folds are yet to form.

We described two important changes in cell shape while discussing

neurulation in amphibians. Such changes occur in chick embryo as well. One
is related to changes in neural ectoderm cells from cuboidal to columnar
shape that results in narrowing and thickening of neural plate. The second is
related to narrowing of the apical ends of some neural epithelial cells when the
neural plate rolls up to form two parallel neural ridges or folds that eventually
meet to form the neural tube.

15.6.1 Mechanisms of Neural Plate Formation

You are now familiar with the concept that the neurulation comprises of two
processes i) the neural plate formation and ii) the neural tube formation. All the
mechanisms responsible for these two processes are not very clear.

Following induction by the underlying mesoderm during gastrulation, the

ectodermal cells that will give rise to the neural tube appears as a thick plate
known as neural plate. 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 may be involved in the elongation process, evidence for this is
inconclusive. The role of microtubules appears to be important in stabilizing
the elongated state of the cells rather than in the elongation process itself.

The bending of neural plate to form neural tube is associated with changes in
cell shape as well. The cells at the edge of the plate are constricted at the
apical end and they become wedge shaped. This change in shape pulls the
edges on both sides up so that the neural plate folds. Cells in the midline of
the neural furrow become wedge shaped that is, the apical edge is constricted
because of the contraction of actin microfilaments and the base becomes
broad. This forms the medial hinge point (MHP). The cells of the MHP are
firmly attached to the notochord which enables the formation of the neural
furrow (Fig.15.14B). Two additional dorsolateral hinge points from where the
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ridge folds curve in to meet (Fig.15.14C). These are anchored to the surface
ectoderm cells. The neural tube is formed as the lateral folds meet in the
midline (Fig.15.14D). When the tube forms, the cells on the apex of the folds
delaminate and fall off as neural crest cells that migrate to their respective
spaces in the embryo.

The neural tube seperates from the presumptive epidermal layer due to
change in cell adhesiveness because neural cells now begin to express N-
cadherin and N-CAM instead of the E-cadherin which is expressed by the
nearby epithelial cells (recall mechanism of cell adhesion from Unit-10). This
allows the neural tube to sink below the surface and the ectodermal cells then
form a continuous layer over it.

Fig.15.14: Stages in the neurulation of chick. A) Neural plate formation; B), C),
and D) The bending of plate in three locations (indicated by arrows),
just above the notochord (N) and on each side of the neural plate,
just below the tips of neural folds

Before we proceed to discuss the mesodermal derivatives, attempt the

following SAQ.

SAQ 5
State whether 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) Neurualtion results in the separation of ectoderm into epidermal
ectoderm, neural ectoderm and neural crest cells.
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 significant role during the rolling up of the neural plate
into neural tube.
vii) In chick the neurulation process occurs simultaneously through out the
length of the embryo.
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Unit 15 Development of Chick
15.6.2 Morphogenesis of Mesodermal Derivatives
Recall from the section on gastrulation that the presumptive mesodermal cells
from the epiblast ingress from the Hensen’s node and from the different
regions of the primitive streak along the dorso-ventral axis into the embryo.to
form the loose sheets of mesodermal cells. 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 the following distinct regions:

Chordamesoderm or axial mesoderm: After the primitive streak is fully

extended in length some cells migrate from the Hensen’s node to move
forwards in the middorsal line. This mesodermal tissue is
chordamesoderm that separates as a middorsal strip from the rest of the
mesodermal tissue. Chordamesoderm gives rise to the notochord which
establishes the anterior-posterior body axis of the embryo. The
notochord gives support to the developing embryo and later is cleared
due to apoptosis remaining only as the intervertebral discs (Fig.15.15B).

Dorsal mesoderm or paraxial mesoderm: As the notochord is laid down,

the mesoderm on each side forms blocks of tissues called somites
present on each side of the neural tube (Fig.15.15C). These will form the
muscles and connective tissue in the back of the embryo, such as
dermis, muscle and skeletal elements (vertebrae and ribs) surrounding
the spinal cord and some muscles in the limb and ventral regions mainly
the abdominal walls. The anterior most paraxial mesoderm does not
form somites and is called head mesoderm. Located in the head region,
it will give rise to head muscles.

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.15.15).

Fig. 15.15: Stages in the development of mesoderm shown in transverse

sections of chick embryo at trunk level. 181
 Block 4 Developmental Biology of Vertebrates-II

Lateral plate mesoderm: Lateral plate mesoderm is a sheet of loosely

connected cells on either side of gut (Fig.15.15 C and D). This loose
sheet of epithelial cells splits into two layers, the somatic mesoderm
which becomes closely associated with ectoderm and the splanchnic
mesoderm which becomes closely associated with endoderm. The
space between the two regions of mesoderm is the future coelomic
cavity (Fig. 15.15D). Lateral plate mesoderm gives rise to heart, blood
vessels and blood cells and the lining of the body cavities. All the
components of pelvic limb skeleton except muscles are derived from
lateral plate mesoderm. Lateral plate mesoderm also helps to form some
of the extraembryonic membranes that are used for transporting
nutrients to the embryo.

SAQ 6
Name the organs derived from

i) Dorsal mesoderm

ii) Lateral plate mesoderm

15.7 FOLDING OF EMBRYO

You have learnt at the beginning of the unit that the entire blastoderm does not
form the embryo. Its central portion, called the area pellucida, only forms the
embryo, and its outer portion, the area opaca forms the extra-embryonic
membranes, such as yolk sac, amnion, serosa (chorion) and allantois.

The body of the embryo becomes separated from the yolk sac. This is
achieved by the formation of folds which appear all around the body of
embryo. The folds involve all three germ layers and are directed downward
and inward, undercutting the body of the embryo proper. They are known as
body folds. The various folds do not appear simultaneously.

The first to appear is the head fold just in front of the head. It undercuts the
head and anterior part of the trunk of the embryo, so that these parts project
freely over the surface of yolk sac. The lateral and posterior parts of the body
folds develop soon after.

The posterior fold undercuts the tail end and the posterior part of trunk which
also projects freely over the surface of yolk sac. These body folds gradually
contract underneath the embryo and eventually the body of embryo is
connected with the yolk sac and other extra-embryonic membranes by a
narrow stalk, the yolk stalk. Figure 15.16 shows the various body infoldings
182 that give it shape and make it constricted off from the yolk.
Unit 15 Development of Chick

Fig.15.16: Post-neurular development. A) Stained preparation of the stage 10;

B) External view of embryo of about 27-28 pair of somites (51 to 56
hours of incubation, stage 16); C) Internal anatomy of stage 16 (of B);
D) Embryo of about 72 to 75 hours of incubation (Stage20); E)
embryo of stage 27, F) Embryo of stage 23.

Flexure and Torsion

When the head is formed, it gets bent down ventrally with respect to the main
axis of the embryo, this bending is called cranial flexure. Then first the head,
and gradually the entire body turns sideways so that the embryo comes to lie
on its left side over the yolk, this dextral twist is knows as torsion. The anterior
end in its cervical region becomes still more bent over towards its ventral
surface.

Various organs are formed on the third and fourth days of incubation, such as
the nervous system, sense organs, four pairs of pharyngeal pouches out of
which only the first three pairs meet the ectoderm to open as gill clefts, five
pairs of visceral arches, heart and, blood vessels, and paired segmental
somites. 183
 Block 4 Developmental Biology of Vertebrates-II

15.8 DEVELOPMENT OF EXTRA-EMBRYONIC

MEMBRANES
As you had seen in Figure 15.15, during neurulation, the lateral plate
mesoderm splits into an outer somatic layer lying beneath the ectoderm and
an inner layer lying outer to the endoderm layer. In between these two layers
of mesoderm lies the coelomic space.

The ectoderm and somatic layer of mesoderm are collectively called

somatopleure, while the splanchnic layer of mesoderm and endoderm form the
splanchnopleure. During later developmental stages, the somatopleure and
splanchnopleure gradually spread outward beyond the developing embryo.

As the body of embryo grows, it becomes separated from the foetal

membranes by the appearance of body folds - cephalic, caudal and lateral
folds. These folds limit the body of embryo and separate the embryonic region
from the extraembryonic region.

The blastoderm besides forming the embryo, gives rise to certain other
structures which do not take part in the formation of embryo proper, but are
external to the developing embryo. These structure are collectively called
extra-embryonic membranes.

These membranes are essential for complete development of the embryo.

These extraembryonic membranes are concerned with nutrition, excretion,
respiration and protection from desiccation and concussions. These
membranes are amnion, chorion or serosa, allantois and yolk sac.

15.8.1 Development of Amnion and Chorion

The origin of amnion and chorion is considered together since they develop
simultaneously from the extra-embryonic somatopleure. Around 30 hours of
incubation, the extra-embryonic somatopleure of the blastoderm rises up in
front of the embryo as a fold, the fold grows and forms an arch over the
embryo.

It forms a double somatopleuric hood, called the cephalic amniotic fold. As this
fold gradually extends backward, its caudally extending side limbs called
lateral amniotic folds arch over the embryo from either lateral side. A similar
fold or elevation over the tail appears, called the caudal amniotic fold. All these
amniotic folds converge and fuse over the embryo, enclosing it within two
sheets of somatopleure from all sides except the region of yolk stalk
(Fig.15.17).

Functions of Amnion and Chorion

The amnion, chorion and amniotic cavity serve the following functions
for the embryo:

a) The amnion protects the embryo from desiccation and sudden

temperature changes.

b) The amniotic fluid is an efficient shock absorber and protects the embryo
from the mechanical shocks.

c) The shell and shell membranes are protective and prevent desiccation of
184 the embryo.
Unit 15 Development of Chick
d) The mesoderm of amnion during later development forms muscle cells
which contract rhythmically, rocking the embryo within the amniotic fluid
so as to prevent it from adhesion to the embryonic membranes.

Fig. 15.17: An early chick embryo showing body folds delimiting from extra-
embryonic areas.

15.8.2 Development of Allantois

About the third day of incubation the floor of the endodermal hindgut begins to
bulge as a bladder, called the allantois. The allantoic evagination is formed
from the splanchnopleure, that is, inner layer of endoderm and an outer layer
of splanchnic mesoderm. It grows rapidly and spreads into the extra-
embryonic coelom, the space between the yolk sac, the amnion and the
chorion.

The distal part of the allantois expands and remains connected with the
hindgut of the embryo by means of a narrow allantoic stalk. When the body
folds contract, embryo is separated from the extra-embryonic parts, the
allantoic stalk is enclosed together with the stalk of yolk sac, or an umbilical
cord is formed (Fig. 15.18).

Fig.15.18: Early stage in the development of the extra-embryonic membranes of

chick.
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 Block 4 Developmental Biology of Vertebrates-II

As the allantois vesicle enlarges and spreads outward, its distal part becomes
flattened and expands between the amnion and yolk sac on one side and the
chorion on the other side. Due to this, the splanchnic mesoderm of the
allantois fuses with the inner somatic mesoderm of the chorion to form an
allanto-chorion.

The allanto-chorion grows around the albumen of the egg to enclose it in an

albumen sac. It aids in absorption of water and albumen. The allantois
becomes highly vascular due to the appearance of blood vessels in the
splanchnopleure. It also receives a pair of allantoic or umbilical arteries, and
its blood is returned into a pair of allantoic or umbilical veins.

The chorio-allantoic circulation continues until the young chick breaks the egg-
shell and begins to breathe the surrounding air. Thus, the umbilical vessels
close, the circulation ceases and the allantois dries up and separates from the
body of the young chick. At the time of hatching, the allantoic vesicle with
excretory wastes is detached from allantoic stalk and is left attached to the
broken shell.

Functions of Allantois

The allantois serves following functions for the embryo:

i) The allantois serves as an embryonic urinary bladder collecting uric acid

from the kidneys which is, thus, prevented from escaping into other parts
of the embryo where it might be harmful.

ii) The vascular allanto-chorion is in contact with shell membranes and it

brings about the respiration of the embryo by an exchange of oxygen
and carbon dioxide through the porous shell.

iii) Development of Yolk-Sac: During neurulation, the gut region of embryo

has a roof and side but no floor. The ventrally open gut rests on the yolk
mass. The formation of yolk sac first among extraembryonic membranes
is essential, since nutrition is supplied to the developing embryo with the
help of yolk sac. The extraembryonic splanchnopleure grows over the
yolk and eventually surrounds the entire yolk to form the yolk sac.

It has an inner layer of endoderm and an outer layer of splanchnic mesoderm.

The yolk sac in the beginning is joined by a wide yolk stalk to the midgut of the
embryo. But later on, as the body folds move toward one another beneath the
embryo, a floor of the gut is formed except in a small region of the midgut
through which it communicates with the yolk sac.

Thus, the yolk sac cavity remains in continuity with the gut cavity through yolk
duct of yolk sac stalk. The endodermal surface of the yolk sac is thrown into
folds called the yolk sac septa that penetrate the yolk mass. The rich blood
circulation develops within the splanchnic mesoderm layer of the yolk sac.
These are the paired vitelline arteries and veins, now called the
186
omphalomesenteric blood vessels (Fig.15.19).
Unit 15 Development of Chick

Fig.15.19: Later stage in the development of the extra-embryonic membranes of

the chick.

The endodermal cells of the yolk sac secrete digestive enzymes which digest
the yolk. The digested yolk is collected by left and right vitelline veins, both of
which open into an unpaired ductus venosus which opens into sinus venosus
of the heart.

Thus, the yolk sac serves as a digestive and absorptive surface by which yolk
is made available to the embryo. Shortly before hatching the shrivelled up
remains of the yolk sac are retracted into the abdominal cavity of the embryo,
and the walls of the abdominal cavity close behind it (Fig.15.20).

Fig.15.20: The fully matured extra-embryonic membranes of the chick.

The amnion, chorion, and allantois are devices by which the embryo can
develop on dry land (as also in reptiles and mammals). The yolk sac (also
found in fishes, reptiles and mammals) develops for the absorption of yolk. 187
 Block 4 Developmental Biology of Vertebrates-II

SAQ 7
Write one important function of each of the following:

i) Amnion

ii) Chorion

iii) Allantois

15.9 HATCHING
The early embryo was at right angles to the long axis of the egg, later it comes
to lie along the long axis with its head near the blunt end. The beak of the
chick is covered by a horny caruncle by which it first pierces the inner shell
membrane to take air into the lungs from the air space. Soon after the
caruncle breaks the egg shell and the chick emerges after 21 days of
incubation at 103°F.

Since the inside surface of shell is concave, it is easier for the chick to break
open the shell and shell membrane as all the force is concentrated at one
point. The outer surface of the shell is convex, hence, the force exerted on the
outside of the shell is distributed on all sides and the shell does not break
easily.

The chick breaks open the shell by repeated blows of the egg tooth and
hatches out leaving behind the allanto-chorion and amnion in the shell. The
newly hatched chick is precocious or nidifugous, i.e., it is fully clad with wet
feathers which soon dry up on the exposure to air, and is able to walk and
feed.

15.10 SUMMARY
Let us summarise what you have learnt so far:

• Chick egg is macrolecithal. It is entirely filled by yolk and yolk is covered

by albumen. Cleavage in chick is discoidal and meroblastic.

• Cleavage produces morula which is converted into blastula or a blastula-

like structure the blastodisc. The organization of blastula is dependent
on the amount of yolk originally deposited in the egg. The amount of yolk
influences the whole course of development and structure of embryonic
stages. Such an influence continues even during subsequent stages of
gastrulation.

• During the gastrulation of aves (also in the reptiles, egg laying

mammals), the primitive streak provides the passage for the movement
of presumptive notochordal, mesodermal as well as endodermal cells.
Hence the primitive streak may be called the closed blastopore of
Amphibia.

• Neurulation consists of, first the neural plate formation and then the
188 neural tube formation. The neural plate is formed by changes in the
Unit 15 Development of Chick
shape of the cells by the cell elongation and accompanying apical
shrinkage. The rolling of neural plate into neural tube due to changes in
cell shape is brought about by the contraction of actin filaments in the
apical end of the cells.

• Post neural development processes include, folding, flexure and torsion

of embryo.

• The blastoderm besides forming embryo gives rise to extra embryonic

membranes i.e. amnion, chorion and allantois. All these perform specific
functions.

15.11 TERMINAL QUESTIONS

1. Discuss the process of gastrulation in chick.

2. Define the following terms:

a) Blastoderm

b) Morphogenetic movements

c) Meroblastic cleavage

d) Fate maps

e) Coelobastula

f) Epiblast

g) Inner cell mass

3. What is the fate of presumptive notochordal area of blastula of the chick

in the adult stage?

4. Describe the process of neurulation in chick.

5. Discuss the process of development of extra embryonic membranes in

chick.

15.12 ANSWERS
Self Assessment Questions
1. a) Macrolecithal b) Blastodisc

2. a) Meroblastic, discoidal

b) When due to cleavage, the blastodisc becomes cellular, round disc

like structure, it is called blastoderm.

3. i) epiblast

ii) ectoderm

iii) area pellucida

189
 Block 4 Developmental Biology of Vertebrates-II

iv) blastocoel

v) polysaccharide

4. i) Convergence, divergence, involution, poly-invagination, epiboly.

ii) Influences formation of antero-posterior medial axis and orientation

of embryo.

iii) Ectoderm, endoderm, mesoderm.

iiii) Individually through primitive streak.

iiv) Extra embryonic membranes (yolk sac, amnion, allantois, chorion).

iv) Refer to Subsection 15.5.7.

5. i) F ii) T iii) T iv) F v) T vi) F vii) F.

6. i) Muscles, cartilage, dermis.

ii) Heart, blood vessels and blood cells.

7. i) The amnion projects the embryo from desiccation and sudden

temperature changes.

ii) The function of chorion is to contribute to the development of the

placenta in placental animals.

iii) The allantois serves as an embryonic urinary bladder collecting

uric acid from the kidneys which is, thus, prevented from escaping
into other parts of the embryo where it might be harmful.

Terminal Questions
1. Refer to Section 15.5.

2. Refer to the relevant parts of the text.

3. The presumptive notochordal area of zygote of a chick forms notochord

in the early embryonic stages but it is destined to be obliterated
completely by the adult stage except in the cyclostomes.

4. Refer to Section 15.6.

5. Refer to Section 15.9.

190