GASTRULATION

AND FORMATION OF THE GERM LAYERS

Gastrulation 1) Invagination: Inpocketing of cells
Ø Characterized by profound but well ordered in at the VP and elongates into
rearrangements of the cells in the embryo the blastocoel
Ø Cells undergo morphogenetic movements 2) Presence of secondary
Type of movement Description Example mesenchymal cells
Invagination Inpocketing of a sheet Archenteron formation o At the innermost tip of
of cells in Amphioxus
Outpocketing of a sheet Exogastrulation
the archenteron
Evagination
of cells o Extend long Filopodia
Involution Rolling around a corner Cell movements toward the opposite wall
of an expanding outer through the amphibian of the blastula as the
layer of cells and blastopore
spreading over an
archenteron elongates
internal surface 3) Tip of archenteron makes contact
Epiboly Spreading of a cell Spreading of outer cells with the blastocoel wall
sheet toward amphibian o Secondary mesenchymal
blastopore
Sinking of individual Primary mesenchyme
cells undergo a major
Ingression
cells from a surface formation in sea urchin determinative event
into an area blastula Ø If original mesenchyme has removed, secondary
Delamination Separation of a second Formation of primary mesenchymal cells transform into primary
(polyingression) sheet from an original hypoblast in avian
single sheet embryos mesenchymal cells
Amoeboid motion Migration of cells as Migration of neural Ø After the contact to the wall of the blastocoel, they
single individuals crest cells (secondary mesenchymal cells) lose capacity and
through their own
begin to migrate and differentiate into 4 types of
motility
cell:
Ø Two main strategies embryos have adopted for
1) Pigment cells
dealing with gastrulation:
2) Blastocoelar cells
1) Carry out gastrulation movements within
3) Two coelomic pouches: protrude
the context of a sphere
from the tip of the archenteron
• See in the embryos of very
4) Circumesophageal musculature
primitive vertebrates (Amphioxus)
Ø Archenteron reaches the opposite wall of the
and amphibians
blastula, bilaminar layer ruptures to from the oral
2) Great amount of yolk dictates gastrulation
opening
• Eggs of reptiles and birds
Ø Blastopore becomes anus
Ø Blastopore – opening from the outside into the
Ø Embryo is becoming a pluteus larva when it begins
archenteron
to a triangular appearance and as the skeleton
Ø Single layer of blastula has been rearranged to
arises from the primary mesenchyme, armlike
form two layers
structures
1) Ectoderm – outer layer
2) Endoderm
• Inner layer
• Upper surface includes cells
destined to from mesoderm
Gastrulation in sea
urchin
Gastrulation in Sea Urchin Embryos
Ø Morphogenetic movements begin in the late
blastula with the separation of the primary
mesenchyme from the wall of the blastula
Ø Filopodia
• Prominent projections that develop from
60 or so ingressed primary mesenchymal
cells
• By extending and retracting, embryos
move along the basal lamina that lines the
blastocoel until they stop and form ringlike
structure near the base of the invaginating
Origin and derivatives
archenteron
of secondary
Ø Unique antigens appear on the surface of the
mesenchyme cells in
primary mesenchymal cells
the sea urchin embryo
Ø Calcified spicules formed on cables serve as the
basis for the internal skeleton of the sea urchin
larva
Ø Archenteron
• Primitive gut
• Occurs in three stages:
 GASTRULATION AND FORMATION OF THE GERM LAYERS
Gastrulation in Amphibian Embryos
Ø Began with the penetration of the sperm into the
egg
Ø Cortical rotation
• Stimulated by sperm fusion
• Leads to the generation of an early
organizing center (Nieuwkoop center) in
dorsal cells of the vegetal hemisphere
• Major activity of this organizing center is
mesodermal induction
Ø Late blastula stage:
• Organizing activity: from original vegetal
organizing center to a more superficial
dorsal location
• Goosecoid and noggin become activated
Ø Blastopore: site at which cells move into the
interior of the embryo
Ø Due to high content of yolk:
• Inward movement of vegetal cells is
impede
• Early Inpocketing of cells is restricted to
the smaller crescent of marginal cells
Ø Dorsal tip of blastopore
• Upper margin of the blastoporal groove
• Later become the major organizing region
(Spemann organizer) of the embryo
Ø Yolk plug – mass of yolk left presenting at the
blastopore
Ø Gastrulation movements are not uniform among
the major amphibian groups
Ø Change in shape of bottle cells is associated with an
inward pulling movement that results in the
formation of the blastoporal groove
Ø Fate maps
• Can summarize the results of many
tracing experiments
• Show on an early embryo the regions that
are destined to give rise to specific
structures or regions later in development
Ø Prospective endoderm
• Around most of the ventral margins of the
blastopore and extending down onto the
ventral part of the embryo
• Rolled into the interior of the embryo and
eventually comes to line its archenteron
Ø Chordamesoderm
• Most of the cells passing over the dorsal lip
of the blastopore
• Give rise to the notochord and cephalic
mesoderm
Ø Involution:
• When begun, the early archenteron is
lined by Chordamesoderm on its dorsal
surface and elsewhere by endoderm
• As it continues, archenteron increases in
size and extends beneath the outer layers
of cells toward the cephalic end
Ø Invaginated endodermal cells spread beneath to
form a complete endodermal lining to the primitive
gut
Ø Gastrulation in Xenopus
• In early dorsal lip: the surface cells are
underlain by a deeper marginal zone
which consist of several layers of cells
• Movement of cells around the dorsal lip of
blastopore are associated with convergence
and extension phenomena
• It provide a motive force
• Also account for the overall elongation of
the embryo that begins during later
gastrulation
• Early gastrulation: cells of the deep layer
of the marginal zone interdigitate by a
process known as radial intercalation to
form a single layer
• Involuted cells that were derived from the
marginal zone will form the mesoderm of
the embryo
Ø Mediolateral intercalation
• A process that begins around the
midgastrula period
• Lateral cells insert processes between cells
located in the medial part of the later and
ultimately become totally interposed
between
Ø The surface layer then expands (epiboly)
Ø As the surface cells pass around the rim of the
dorsal lip, they become the endodermal lining of the
archenteron
Ø Bottle cells decrease in height
Ø One region is destined to take part in the formation
of CNS and is therefore designated as neural
ectoderm
Ø Remainder of the ectoderm will form the epidermis
and therefore called general cutaneous ectoderm
Ø Spemann and Mangold
• Demonstrated that the dorsal lip of the
blastopore possesses the ability to
“organize” much of the future development
of the embryo
• Spemann called the dorsal lip blastopore
the organizer
Caudal view of
amphibian embryo
Bottle cells
 GASTRULATION AND FORMATION OF THE GERM LAYERS
Prospective areas of amphibian embryos at the stage when
gastrulation is starting
Cell organization in
amphibian
gastrulation
Gastrulation in Birds
Ø Blastula (in chick embryo):
• Epiblast: upper layer
• Primary hypoblast: lower layer
• Blastocoel: in between
• Area pellucida: transparent
• Area opaca: surround area pellucida and
where the cells of the blastoderm lie
unseparated from the yolk
Ø Koller’s sickle
• Thin sickle-shaped mass of cells
• Takes shape at the posterior end of the
embryo
Ø Secondary hypoblast
• Pushes anteriorly, compressing and folding
the primary hypoblast ahead of it
• Forms extraembryonic endoderm,
principally the yolk stalk
Ø Neither the primary nor the secondary hypoblast
seems to form any of the embryonic germ layers
Ø Primordial germ cells are found in the primary
hypoblast
Ø Gastrulation and formation of the definitive
embryonic germ layers begin with the appearance
of a condensation of cells in the posterior part of the
epiblast
Ø Incubation of 3-4 hours: condensation gradually
assumes a cephalocaudal elongation
Ø Incubation of 7 or 8 hours: elongation is still
definite
Ø End of the first half day: thickened areas has a
shape which has led to being the primitive streak
Ø Primitive streak
• Result of an inductive interaction of the
epiblast with the hypoblastic layer
• Early primitive streak: elongates in both a
cephalic and a caudal direction
• 16 hours of incubation: primitive streak
stage
• Primitive groove – central furrow that
runs down the center of the primitive
streak
• Primitive ridges – a thickened margins in
both sides of primitive groove
• Hensen’s node – a local thickening at the
cephalic end of the primitive streak
Ø Incubation of 18 hours: primitive streak has
reached its full length and the cephalic end begins
to regress
Ø Head process – the area where the notochord has
been recently laid down
Ø Embryonal area or embryonic shield – form when
the part of the area pellucida adjacent to the
primitive streak begins to thicken
Ø Long axis of the future embryonic body is
established by primitive streak
Ø Embryonic germ layers are formed by migration of
cells in the epiblast toward Hensen’s node and the
primitive streak
Ø Their ingression forms the embryonic mesoderm
and endoderm
Ø Anterior portion of the primitive streak and node
serve as a passageway for cells
Ø Future embryonic endodermal cells: first cells to
pass through the area of the anterior part of the
primitive streak
Ø After 8-10 hours of incubation, more than 80% of
cells are found in the endoderm
Ø Endodermal cells then enter the original
hypoblastic layer and displace the cells of the
hypoblast outward and cephalad toward the edge of
the area opaca
Ø 22 hours of incubation: primitive streak regression
has commenced, all future endodermal cells have
left the epiblast
Ø Major sites of invagination & mesodermal
formation:
a) Along the length of the primitive streak
• Coherent layer of mesodermal cells
expands parallel to the underlying
layer of the embryonic endoderm
b) Through Hensen’s node
• A rod of mesodermal cells directed
cephalad lies in the midline of the
embryo
• Mesodermal rod becomes the
notochord
Ø Mesodermal cells that exit the cranial part of the
primitive streak > formation of embryonic
mesoderm
Ø Mesodermal cells that exit from the caudalmost
part of the primitive streak > part of the
extraembryonic mesoderm
Ø Epiblast
• Cuboidal to columnar epithelial cells
 GASTRULATION AND FORMATION OF THE GERM LAYERS

• Deeper parts are bound together by gap Ø Prospective potency – the type of differentiation of
junctions which a cell or group of cells is capable at a given
• When cells enter the primitive groove, it stage of development
undergo change of shape Ø Typically, prospective potency > prospective
• Change of shape is associated with the significance (at early developmental stages)
appearance of intracellular microtubules Ø As developmental restriction sets in, prospective
• Tight junctions begin to break up potency = prospective significance
Ø Techniques used in the construction of fate maps:
a) Mark certain cells with vital dyes
b) Transplantation of radioactively labeled
pieces of early embryos into equivalent
locations in unlabeled host
c) Microinjection into individual cells of
intracellular dyes or molecular marker
d) Explanting small pieces of embryos as
grafts onto the chorioallantoic membrane
or into the coelomic activity
Ø Prospective center – territory where a specific
potency is regularly manifested
Ø Cell adhesion molecules:
Primitive streak • Pregastrulation: epiblast and hypoblast
stage contain N-CAM and L-CAM
• As mesodermal cells migrate through the
Four stages in the formation primitive streak, neither CAM can be
of Primitive Streak detected
• Cells of future nervous system lose their L-
CAM but retain N-CAM
• Nonneural ectoderm retain L-CAM but
lose N-CAM
Formation of
the mesodermal
layer in early chick
embryo

Map of the
prospective areas of
the outer layer of the
chick embryo in the
primitive streak

Molecular Aspects of Early Avian Development

Ø Expression of some embryonic genes begins as
early as the morula stage
Ø Activin is expressed in the hypoblast and can
induce axial structures (notochord, somites and
neural tube) in explants or epiblast
Ø Parallel with the mesodermal induction by specific
vegetal region in amphibian embryo
Ø Goosecoid genes is expressed in Hensen’s node and Map of the
even in cells of Koller’s sickle prospective areas of the
Ø Parallel with the pattern of expression of this gene invaginated layer of
in amphibian embryo the chick in the
Ø Retinoic acid primitive-streak stage
• An important morphogen
• Hensen’s node is rich with this
Prospective Areas in Chicks at the Primitive-Streak
Stage
Ø Prospective significance (prospective fate) – the fate
of a cell or a group of cells during the course of
normal development
 GASTRULATION AND FORMATION OF THE GERM LAYERS
Comparison of Avian and Amphibian Development
Bird embryo Amphibian embryo
Two-layered structure Surface layer of the embryo
containing epiblast and contains cells destined to
hypoblast (blastula stage) become part of the endoderm
Cells that will form the and mesoderm
endoderm and mesoderm are
located in the surface layer of
the embryo
Effects of the hypoblast on the Induction of mesoderm and
epiblast in early chick embryos control of polarity by the vegetal
yolk mass
Movement of surface cells Cells move by means of epiboly
toward the primitive streak in toward the amphibian
the chick blastopore
Cells that will constitute the future endodermal layer migrate from
exterior to the interior of the embryo first and the inward
movement of the mesodermal cells follow later
Area where the chick notochord Dorsal lip of blastopore where
is formed the amphibian notochord arises
Origin of the germ layers in Mammals
Ø Morphogenetic movements and tissue displacement
are similar with birds
Ø Inner cell mass can be compared with the cap of
blastomeres situated in the AP of the yolk sphere
Ø Hypoblast
• A thin layer formed by the first cells that
segregate out from the inner cell mass
• Forms only extraembryonic endoderm
• Equivalent to the hypoblast of the chick
embryo
• Contributes the cells that will line the yolk
sac
Ø Epiblast
• Remainder of the inner cell mass
• Future ectodermal cells
• Contains the cell that will ultimately
migrate through the primitive streak and
become the definitive endoderm and
mesoderm of the embryo
Ø Embryonic disk
• Collective term for the remaining cells of
the inner cell mass that become more
regularly arrange
• Soon, one margin of the disk becomes
thickened
• The thickening part is destined to become
the caudal end of the embryo
• This caudal thickening results to the
formation of the primitive streak
Ø Cells that constitute the embryonic mesoderm and
some extraembryonic mesoderm appear to pass
through the posterior part of the primitive streak
Ø Earliest extraembryonic mesodermal cells originate
from the endoderm (hypoblast) of the yolk sac and
not the trophoblast
Ø Embryonic mesoderm
• Arises by the migration of cells in the
epiblast toward the primitive streak
Ø Nodal
• Member of TGF-β gene family
• Expressed around Hensen’s node
• Encode a secreted signaling molecule that
is essential for the formation of mesoderm
• If expression of this gene is blocked,
mesoderm fails to form
Origin of the germ layers in Rodents
Ø Primitive endoderm
• Early hypoblast
• Forms a single layer beneath the inner cell
mass
• Cells spread out beneath the trophoblast
(trophectoderm) to form the endodermal
layer of the parietal yolk sac
Ø Reichert’s membrane
• A thick basement membrane secreted by
the parietal endodermal cells
• Served as a source of material for
analytical studies of basal laminae
Ø Sections of trophectoderm:
a) Polar trophectoderm
• The part overlying the inner cell
mass
• Cells are still able to undergo
mitotic division
• Descendants contribute to the
mural trophectoderm
b) Mural trophectoderm
• Remainder; surrounding the
blastocyst cavity
• Cells are unable to undergo
normal mitotic divisions
• Cells then transformed into giant
cells by becoming polypoid
Ø Inner cell mass protrudes deeply into the blastocyst
cavity in the form of a tonguelike lobe
Ø Proamnion
• A cavity that forms within the lobe
• Cells surrounding it represent the
primitive ectoderm
Ø Inverted egg cylinder – term for the embryo at this
stage due to its unusual configuration
Ø Ectoplacental cone – above the primitive ectoderm;
form from the cells of the polar trophoblast
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