DEVBIOL

LECTURE 5: GASTRULATION
Gliceria B. Ramos| 1st TERM | A.Y. 2022-2023

OUTLINE ○ Starts with establishing the DLB, to start the

I. A IV. B
formation of the opening, the blastopore
II. ○ Is marked by the formation of the blastopore
● 2. Cell movement and rearrangements
(morphogenetic movements) - prelude to
morphogenesis (gradually generating the overall body
plan)
○ Every step is crucial
○ Through these movements the embryonic
cells are gaining molecular cues
RECAP OF BLASTULATION ● 3. Starting to acquire positional information
● Amphibian ○ PI: molecular cues that will tell an embryonic
○ At the end of blastulation: cell where it is relative to the body axis
■ embryo is a hollow ball of cells with ○ Molecular cues are collectively known as
a cavity, blastocoel positional information (PI)
■ Established dorsal side, the grey ● 4. Formation of the 3 germ layers:
crescent area in the marginal zone ○ Ectoderm - outermost
■ Polarity is established early ○ Mesoderm - middle
● Avian ○ Endoderm - innermost
○ At the end of blastulation:
■ Resulting stacks of cell layers
(epiblast and hypoblast)
● Formation of these layers
mark the early phase of
gastrula
● Swift transition from late
blastula to early gastrula
■ Before these layers, massives cells THREE GERM LAYERS
are formed in what used to be the
blastodisc Epithelial Type Mesenchymal type
■ When the blastodisc cleaves it
becomes the blastoderm Ectoderm and Endoderm Mesoderm
■ Yellow yolk - oocyte/egg cell Flat sheet, closely packed Loosely arranged
● Blastoderm is the center cells
■ Cells of blastoderm start to fall off, Little amount of ECM Plenty of ECM
and become the hypoblast
■ When the layers split, they become Examples: simple Examples: neural crest
the blastocoel, separating the squamous cells of cheeks cells and head
epiblast and hypoblast and stratified squamous mesenchymal cells
■ Subgerminal cavity epithelial cells of the
● Under blastoderm epidermis
● Formed before the two -Spread -Migrate
layers and blastocoel -Roll -Intercalate
● Increasing number of -Fold -Ingress (Movement from a
blastoderm absorb fluid in -Buckle flat sheet layer of cells into
yolk under, creating the -Bend a cavity)
cavity
● Not yet the blastocoel
■ Polarity is established early
● Site of the first falling of
cells that for the hypoblast
is the posterior end
● Mammals
○ At the end of blastulation: Can undertake major Can undertake changes in
■ 2 distinct populations of cells: inner morphologic movements: cell behaviors/activities:
cell mass (ICM) and the trophoblast - Invagination - Migration
(and the blastocoel) - Epiboly - Intercalation
■ Embryo proper is not yet formed in - Involution - Change in cell
ICM - Convergent shape
■ No polarity established yet extension - Change in cell
● Most labile - Delamination adhesiveness
- Passive movement - Change in the rate
of cell division
- Ingression
- Apoptosis

● The germ layers have the ability to perform

epithelial-mesenchymal transition while in
embryonic stage
○ Behaviors and cellular activities happen in
various combinations
○ Regulate by gene activity/activities
■ Genes are activated so the
GASTRULATION products can facilitate the
● Blastula → early gastrula → late gastrula morphological movements and
● Dorsal lip of blastopore cellular activities
○ Landmark of early gastrula ■ Examples: expression of genes for
● Dorsal lip of blastopore becomes well formed by the cyclins and cd kinases (regulate cell
late gastrula stage division)
○ Surrounding the complete blastopore ■ Expression of regulatory genes
○ 3 primary germ layers are formed (code for transcription factors) for
● “Gaster” Latin word- means stomach activation of other genes
● 1. Main goal: establish precursor of digestive gut, the ● Upregulate or
primitive gut (archenteron) downregulate genes

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 ■ Selector genes control the ○ Two or more rows of cells intercalate, but the
expression of CAMs and SAMs intercalation is highly directional
(substrate adhesion molecules) ○ Cells converge by intercalating perpendicular
(morphoregulatory molecules of to the axis of extension
embryonic development) ○ Result: overall extension of the tissue in a
● Examples of combinations: preferred direction
○ Change in rate of cell division (rapid) → cells ○ Intercalation - meet in midline, lateral or
spread out (epiboly) radial
○ Mass movement of cells and involution

MORPHOGENETIC MOVEMENTS
● Invagination
○ An epithelial sheet bends inward to form an
inpocketing
○ Formation of (dorsal lip of) blastopore
○ Only a few cells are involved (localized)
○ Example: localized depression on balloon
when you pinch your thumb

○ The process by which tissue elongates along

the anterior-posterior (AP) axis, and
becomes narrower along the medio-lateral
● Epiboly (ML) axis
○ A sheet of cell spreads by thinning, that is,
the sheet thins, while its overall surface area
increases in the other two directions
○ Can involve a monolayer (i.e. a sheet of cells
one cell layer thick) in which case the indiv.
cells must undergo change in shape
○ Faster cell division in animal pole due to lack
of yolk
○ Mitosis ○ Convergent thickening - involves the tissue
becoming thicker in the direction at right
angles to the convergent extension

● Involution
○ In contrast with invagination
○ Mass movement of cells rolling inward to
form an underlying layer via bulk movement
of cells
○ Example: Ingredients on the top of the
halo-halo will move to the bottom of the tall
glass when you put in your spoon
Sketch showing the convergence force (y-axis) increases
through gastrulation, and the plateaus (during early
neurulation) before increasing again (during late neurulation)
● Delamination
○ Splitting of layer of cells
■ Cells falling off from the thick layer

● Passive movement and Migration of Cells

● Convergent extension ○ Amoeboid Migration
■ (e.g. Primordial germ cells)

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 ■ Migration by amoeboid strength of the adhesion contact
motion between two cells.
○ Mesenchymal Migration ■ At tissue level, Cortex tension and
■ (e.g. neural crest cells & cell-cell adhesion determine the
head mesenchyme) shape of multicellular aggregates
● Powerful and the sorting order in heterotypic
migratory cells aggregates
○ Collective migration ■ At embryonic level, The interaction
■ (e.g. gastrulating cells) between tissues, forming at
● Involution inward different phases of development
○ These cellular activities are and characterized by different
regulated by gene activity cortical and adhesive properties,
controls correct germ layers
formation during gastrulation.

● CAMs and SAMs (Morphoregulatory molecules) →

capable of reversible adhesion; quick
attachment/detachment
● Ingression ● Embryonic cells synthesize stage-dependent and
○ Cells leave an epithelial sheet by region-specific ECM components in accord with
transforming to mesenchymal cells cell activities (spatio-temporal gene expression)
to move into a cavity ○ CAMs (cell adhesion molecules)
■ Cell to cell contact
■ Example:
● Compaction of
blastomeres in mammalian
blastocyst
● Remember compaction (at
8-cell stage) in mammals -
CAMs are involved
■ Undergo dynamic expression
patterns correlated with cell fates
● Examples:
○ E-cadherin - high
expression in
prospective
epidermis
● Change in cell shape and position ○ N-cadherin -
○ (e.g. blastocyst of mammalian embryo) high expression
during 16-cell stage, there is polarization in prospective
■ Polarized outside = trophectoderm neural plate
■ Polarized inside = inner cell mass ○ SAMs (substrate adhesion molecules)
○ E.g. Formation of bottle shaped cells ■ Cell to ECM contact
(epithelial type cells) in amphibian blastopore ■ Establish structural
for involution integrity/relationship between cells
■ Epithelial cells have basement and and the surrounding matrix (ECM)
apical membrane, apical surface is ■ Provide contact guidance to
unattached unlike basement migrating cells
membrane, thus, when they change ■ Examples:
in cell shape, they pull along with ● Laminin, fibronectin,
them the basement membrane to integrins (present in ECM)
eventually form the blastopore

● Apoptosis ● The process of morphoregulation (concept proposed

○ Programmed cell death
○ E.g. Deletion of webbing in digits
● Change in degree of cell adhesiveness
○ Acquire different degrees from cell level,
tissue level, and embryonic level
■ interface-specific localization of
cadherins and the actomyosin
cortex determine the shape and the
by Edelman) that regulates morphogenesis during
development, adaptation, and regulation involves
cellular programmes such as cell division, movement,
adhesion and death, and is controlled by molecules.
● The coordinated expression and function of these
morphoregulatory molecules (CAMs: cell adhesion
molecules; SAMs: substrate adhesion molecules;
JAMs: cell junctional molecules) provide an
essential link between genetic and epigenetic
mechanisms

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● These molecules exert critical interactions at both the
cell surface and the cytoskeleton and mediate their
effects through activation of intracellular signalling
cascades such as the p21ras/MAP-kinase pathway
● Role of CAMs and SAMs in morphoregulatory
cycles
○ Selector genes: called for transcription
factors (transcription control)
○ CAMs/SAMs genes: controlled by
transcription factors depending on embryonic
stage and region at a specific time; then
synthesized and deployed
○ Cell-cell adhesion: interaction with
CAM/SAM of other cells/ECM; cause various
cell movements and may involve changes in
cell shape (drivers of morphogenesis)
○ Morphogenesis: new cell-to-cell signaling
(intracellular) to provide feedback to selector
genes if more CAMs/SAMs genes are
needed
○ Cycle will be repeated as long as more
CAMs/SAMs are needed for embryonic
positioning
Amphibian Gastrulation
● At the start of amphibian gastrulation, there is
orchestration of the 3 morphogenetic movements —
invagination, epiboly, and involution.
● Gray Crescent
○ It determines the site of the dorsal lip of the
blastopore (DLB)
● Dorsal Lip of Blastopore
○ Formed due to bottle-shaped cells
○ Undergo invagination
● The cells in animal hemisphere
○ Undergo mitotic division — epiboly
○ With continuous spreading of cell, goes
inward — involution
■ This can intercalate with the
ectoderm, others will also
intercalate with the endoderm —
eventually causing the elongation of
the whole embryo in the late
gastrula stage.
● Dorsal Lip of the Blastopore (RED PART)
○ This now becomes the site of cell turnover
■ Continuous involution – cells
continuously go inward
○ Cells migrate from DLB to the other side
● Migration of cells
○ It is guided by Fibronectin
○ Anteriorly
● Chordamesoderm
○ Pick-up point for neurulation
○ The last group of cells at the vicinity of DLB
that cannot enter the other side will form
Chordamesoderm
○ This has a powerful influence on the
formation of the neural tube
Avian Gastrulation
● Blastodisk cleaves after fertilization
○ This forms eventually as Blastoderm
○ Blastoderm will start to absorb the fluid
underneath — creating a subgerminal cavity
● Subgerminal cavity forms underneath the blastoderm
● Blastoderm delaminates to form epiblast and primary
hypoblast
● Blastocoel forms at the expense of the subgerminal
cavity
● Blastoderm
○ Top view of the yolk
○ Have undergone cleavage division
○ There is a subgerminal cavity underneath —
viewed under a microscope
■ Black area - blastoderm
■ Light yellow area - area pellucida
(lucid - translucent)
■ Dark yellow area - area opaca (has
close contact with underlying yolk)
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● Blastoderm (Sagittal section)
○ Eventually, splitting into hypoblast and
epiblast — creating the blastocoel (that is
formed in expense of subgerminal cavity)
○ Blastocoel
■ Site where the first falling of cells
will occur — the thickening
● Primitive streak
○ Forms at the epiblast
○ This will designate the future posterior end
and future anterior end of the embryo
● Sagittal section
○ Area pellucida
■ Translucent region occupied by the
blastoderm and the subgerminal
space
○ Area opaca
■ Opaque region because of the
close contact with the underlying
yolk
■ Its cells involved in the processing
of the yolk
● Cells delaminate and ingress from the epiblast into
subgerminal cavity
○ Involves falling off of cells (ingression)
● Eventually, the ingressing cells from the epiblast will
form the primary hypoblast
● Cells from posterior margin (secondary hypoblast)
migrate anteriorly and join the primary hypoblast
Avian gastrulation:
Epiblastic cells move into the cavity
● cells at the anterior end are already starting to form
organs
● posterior portions of the embryo (from HN) are still
undergoing gastrulation
● The primitive streak with the HN will form the
posterior end of the digestive gut, the anus
● Cells anteriorly now starts to form organs, or they
start undergoing neurulation
● The other half, which is more posteriorly: Hensen’s
node is still undergoing late gastrulation, and yet, it is
now regressing posteriorly; Pushed downward
posteriorly is the primitive streak
● The neural structures are pushing anteriorly
● Basically, the neural structures are pushing anteriorly,
and these are all found in the anterior part, whereas
in the posterior part, everything would be the
primitive structures
● If we trace further down, it will now regress into the
posterior end, which forms the posterior end of the
digestive gut that will eventually give rise to the anus.
● All of these give rise to the organs that start out with
the neural structures
● Hensen’s node will be regressing downwards
● From the point posterior the Hensen’s node and
downwards, would be the primitive structures
● From the point anterior to Hensen’s node and
upwards, are the neural structures
Proamnion
- is on top of the merging point of the ectoderm (surface) and
migrating cells underneath which is the endoderm
- This is devoid of mesenchyme, so it appears lighter
- this is within the area pellucida
Notochord
- will form the scaffold
- It provides an axial skeleton for the formation of the CNS, but
it will eventually undergo degeneration, and will be
incorporated into the vertebral column as the nucleus
pulpusos
● The area occupied by the embryo is the area
pellucida and outside is the area opaca
`
Mammalian Gastrulation
- Marked with the formation of a primitive streak similar
with that of avian embryo
● Mammalian gastrulation is the same as in the avian
embryo on establishing the primitive streak
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 ● The inner cell mass will undergo delamination, and it
will form the epiblast and hypoblast

● The epiblast will eventually give rise to the amniotic

ectoderm, which encloses an amniotic cavity

Epiblast will split into two:

1. amniotic ectoderm, which encloses a cavity called the
amniotic cavity
2. remaining or remainder of the embryonic epiblast, which is
where the primitive streak will form.

● Formation of the epiblast and hypoblast is bilaminar

germ disc formation
● Primitive streak is here
● Gastrulation is complete

● Embryonic epiblast is where the primitive streak will

form and there will be ingression of cells
● Those that are ingressing will form the embryonic
ectoderm and embryonic mesoderm
● Left on top, or the outermost would be forming the
embryonic ectoderm
● Three primary germ layers are formed and this is
trilaminar disc formation
● The amniotic ectoderm and the rest of the hypoblast
and trophoblast will contribute to the large part of the
placenta

PLACENTATION

● Primitive streak forms in the epiblast ; it involves

ingression
● Endoderm forms
● Mesodermal cells form
● The ones left at the midline will form the notochordal
process which moves anteriorly ; notochordal process
is the same as that of the axial mesoderm