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 • In early dorsal lip: the surface cells are
Ø Began with the penetration of the sperm into the underlain by a deeper marginal zone
egg which consist of several layers of cells
Ø Cortical rotation • Movement of cells around the dorsal lip of
• Stimulated by sperm fusion blastopore are associated with convergence
• Leads to the generation of an early and extension phenomena
organizing center (Nieuwkoop center) in • It provide a motive force
dorsal cells of the vegetal hemisphere • Also account for the overall elongation of
• Major activity of this organizing center is the embryo that begins during later
mesodermal induction gastrulation
Ø Late blastula stage: • Early gastrulation: cells of the deep layer
• Organizing activity: from original vegetal of the marginal zone interdigitate by a
organizing center to a more superficial process known as radial intercalation to
dorsal location form a single layer
• Goosecoid and noggin become activated • Involuted cells that were derived from the
Ø Blastopore: site at which cells move into the marginal zone will form the mesoderm of
interior of the embryo the embryo
Ø Due to high content of yolk: Ø Mediolateral intercalation
• Inward movement of vegetal cells is • A process that begins around the
impede midgastrula period
• Early Inpocketing of cells is restricted to • Lateral cells insert processes between cells
the smaller crescent of marginal cells located in the medial part of the later and
Ø Dorsal tip of blastopore ultimately become totally interposed
• Upper margin of the blastoporal groove between
• Later become the major organizing region Ø The surface layer then expands (epiboly)
(Spemann organizer) of the embryo Ø As the surface cells pass around the rim of the
Ø Yolk plug – mass of yolk left presenting at the dorsal lip, they become the endodermal lining of the
blastopore archenteron
Ø Gastrulation movements are not uniform among Ø Bottle cells decrease in height
the major amphibian groups Ø One region is destined to take part in the formation
Ø Change in shape of bottle cells is associated with an of CNS and is therefore designated as neural
inward pulling movement that results in the ectoderm
formation of the blastoporal groove Ø Remainder of the ectoderm will form the epidermis
Ø Fate maps and therefore called general cutaneous ectoderm
• Can summarize the results of many Ø Spemann and Mangold
tracing experiments • Demonstrated that the dorsal lip of the
• Show on an early embryo the regions that blastopore possesses the ability to
are destined to give rise to specific “organize” much of the future development
structures or regions later in development of the embryo
Ø Prospective endoderm • Spemann called the dorsal lip blastopore
• Around most of the ventral margins of the the organizer
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 Caudal view of
of the blastopore amphibian embryo
• 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 Bottle cells
gut
Ø Gastrulation in Xenopus
 GASTRULATION AND FORMATION OF THE GERM LAYERS

• 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
Prospective areas of amphibian embryos at the stage when • Hensen’s node – a local thickening at the
gastrulation is starting 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
Cell organization in primitive streak begins to thicken
amphibian Ø Long axis of the future embryonic body is
gastrulation 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
Gastrulation in Birds serve as a passageway for cells
Ø Blastula (in chick embryo): Ø Future embryonic endodermal cells: first cells to
• Epiblast: upper layer pass through the area of the anterior part of the
• Primary hypoblast: lower layer primitive streak
• Blastocoel: in between Ø After 8-10 hours of incubation, more than 80% of
• Area pellucida: transparent cells are found in the endoderm
• Area opaca: surround area pellucida and Ø Endodermal cells then enter the original
where the cells of the blastoderm lie hypoblastic layer and displace the cells of the
unseparated from the yolk hypoblast outward and cephalad toward the edge of
Ø Koller’s sickle the area opaca
• Thin sickle-shaped mass of cells Ø 22 hours of incubation: primitive streak regression
• Takes shape at the posterior end of the has commenced, all future endodermal cells have
embryo left the epiblast
Ø Secondary hypoblast Ø Major sites of invagination & mesodermal
• Pushes anteriorly, compressing and folding formation:
the primary hypoblast ahead of it a) Along the length of the primitive streak
• Forms extraembryonic endoderm, • Coherent layer of mesodermal cells
principally the yolk stalk expands parallel to the underlying
Ø Neither the primary nor the secondary hypoblast layer of the embryonic endoderm
seems to form any of the embryonic germ layers b) Through Hensen’s node
Ø Primordial germ cells are found in the primary • A rod of mesodermal cells directed
hypoblast cephalad lies in the midline of the
Ø Gastrulation and formation of the definitive embryo
embryonic germ layers begin with the appearance • Mesodermal rod becomes the
of a condensation of cells in the posterior part of the notochord
epiblast Ø Mesodermal cells that exit the cranial part of the
Ø Incubation of 3-4 hours: condensation gradually primitive streak > formation of embryonic
assumes a cephalocaudal elongation mesoderm
Ø Incubation of 7 or 8 hours: elongation is still Ø Mesodermal cells that exit from the caudalmost
definite part of the primitive streak > part of the
Ø End of the first half day: thickened areas has a extraembryonic mesoderm
shape which has led to being the primitive streak Ø Epiblast
Ø Primitive streak • 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 • If expression of this gene is blocked,
Bird embryo Amphibian embryo mesoderm fails to form
Two-layered structure Surface layer of the embryo Origin of the germ layers in Rodents
containing epiblast and contains cells destined to Ø Primitive endoderm
hypoblast (blastula stage) become part of the endoderm
• Early hypoblast
Cells that will form the and mesoderm
endoderm and mesoderm are • Forms a single layer beneath the inner cell
located in the surface layer of mass
the embryo • Cells spread out beneath the trophoblast
Effects of the hypoblast on the Induction of mesoderm and (trophectoderm) to form the endodermal
epiblast in early chick embryos control of polarity by the vegetal layer of the parietal yolk sac
yolk mass Ø Reichert’s membrane
Movement of surface cells Cells move by means of epiboly • A thick basement membrane secreted by
toward the primitive streak in toward the amphibian
the parietal endodermal cells
the chick blastopore
• Served as a source of material for
Cells that will constitute the future endodermal layer migrate from
exterior to the interior of the embryo first and the inward analytical studies of basal laminae
movement of the mesodermal cells follow later Ø Sections of trophectoderm:
Area where the chick notochord Dorsal lip of blastopore where a) Polar trophectoderm
is formed the amphibian notochord arises • The part overlying the inner cell
mass
Origin of the germ layers in Mammals • Cells are still able to undergo
Ø Morphogenetic movements and tissue displacement mitotic division
are similar with birds • Descendants contribute to the
Ø Inner cell mass can be compared with the cap of mural trophectoderm
blastomeres situated in the AP of the yolk sphere b) Mural trophectoderm
Ø Hypoblast • Remainder; surrounding the
• A thin layer formed by the first cells that blastocyst cavity
segregate out from the inner cell mass • Cells are unable to undergo
• Forms only extraembryonic endoderm normal mitotic divisions
• Equivalent to the hypoblast of the chick • Cells then transformed into giant
embryo cells by becoming polypoid
• Contributes the cells that will line the yolk Ø Inner cell mass protrudes deeply into the blastocyst
sac cavity in the form of a tonguelike lobe
Ø Epiblast Ø Proamnion
• Remainder of the inner cell mass • A cavity that forms within the lobe
• Future ectodermal cells • Cells surrounding it represent the
• Contains the cell that will ultimately primitive ectoderm
migrate through the primitive streak and Ø Inverted egg cylinder – term for the embryo at this
become the definitive endoderm and stage due to its unusual configuration
mesoderm of the embryo Ø Ectoplacental cone – above the primitive ectoderm;
Ø Embryonic disk form from the cells of the polar trophoblast
• 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 *HANGGANG DITO NA LANG!!!! DI KO NA KERI*
formation of the primitive streak
Ø Cells that constitute the embryonic mesoderm and *FOR THE LATTER PARTS, TINGNAN NIYO NA LANG
some extraembryonic mesoderm appear to pass PICTURES SA BOOK HEHEHEHE*
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