Gastrulation

• Definition = migration and division of cells to set up the 3 primary

germ layers.
• What positions do presumptive germ layers occupy in the blastula,
before gastrulation?
– Experimental answers to this question provide our basis for
understanding cell movements that occur during gastrulation.
• FATE MAPS = diagram of blastula/blastodisc showing the “fate” of
each part.
• Fate mapping technique developed by Vogt in 1920s. Involves …
– Marking surface of blastula with vital dyes
– Dyes retained by cells for prolonged periods, but don’t interfere with
normal cellular processes
– Follow movements of marked cells during gastrulation to ultimate
locations in later embryos
Sample fate map of frog embryo
 Gastrulation
• Micro- and Mesolecithal eggs (Amphioxus and
Amphibians) …
• Basic cell movements during gastrulation are
spreading and stretching, accompanied by cell
division.
• 7 stages to gastrulation in micro- and
mesolecithal eggs.
 Steps in Gastrulation
1. Prospective endodermal cells in vegetal hemisphere elongate and move
toward interior.
2. Base of cell remains attached to surface of blastula, this causes strain on
surface which causes an indentation to form (blastopore).
3. Area directly above the blastopore = Dorsal Lip of Blastopore. This is
where cells begin to move inward.
4. Invagination spreads circulolaterally enclosing yolk plug of endoderm.
5. Continued invagination and spreading so that surface of gastrula
contains only ectoderm.
6. Notochord and somatic mesoderm become stretched out longitudinally
inside, neural ectoderm longitudinally on surface.
7. When complete:
• composite inner layer involving both mesoderm and endoderm
• notochordal mesoderm and neural ectoderm in parallel
• blastocoel eliminated by spreading of endoderm
• yolk plug has withdrawn internally so blastopore is only a narrow slit
Fig 5.7 – Gastrulation and
Neurulation in Amphioxus
Fig 5.8 – Early embryonic development in Lampreys
Fig 5.11 – Gastrulation
and Neurulation in
Amphibians
 Amphibian Gastrulation Video
http://worms.zoology.wisc.edu/frogs/gastxen/wholegas.html
Gastrulation in Macrolecithal Eggs
• Pattern of movements is different, but the
process is basically the same.
• Elasmobranchs = basically infolding over
posterior margin of blastodisc.
• Outer surface becomes ectoderm, inner
surface endoderm, mesoderm resides in
between.
• Entire embryo still on top of yolk.
 Gastrulation in Macrolecithal Eggs
Reptiles and Birds:
1. Blastodisc contains cells of varying size: large, yolk-rich cells accumulate
at lower and posterior surface, small yolk-poor cells at upper surface.
2. A separation gradually forms between large lower and small upper cells
(= blastocoel), forms epiblast (dorsally) and hypoblast (ventrally).
– Upper epiblast forms prospective ectoderm and mesoderm, lower hypoblast
forms prospective endoderm.
3. Hypoblast spreads anterolaterally to form endoderm layer.
4. Mesoderm infolding occurs in central region of embryo.
– Infolding begins at posterior end of blastodisc; progresses anteriorly forming a
groove bounded by parallel ridges = primitive streak
– At anterior end of this groove, there exists a raised node of tissue with a pit
(primitive pit) extending down and forward beneath it. Node = Henson’s Node –
equivalent of dorsal lip of blastopore in micro- and mesolecithal eggs.
5. Mesodermal tissue migrates downward along primitive streak
expanding laterally beneath ectoderm and above endoderm.
Fig 5.13 – Gastrulation in the
Bird embryo
Chick Gastrulation Video
http://www.gastrulation.org/
Gastrulation in Macrolecithal Eggs
• Hypoblast may exert causal influence on
primitive streak:
• If hypoblast separated from epiblast and
reoriented, the axis of the primitive streak is
similarly reoriented.
• If hypoblast removed, primitive streak fails to
form
• These experiments suggest that the hypoblast
forms a necessary substratum for orientation
and deployment of prospective mesoderm.
 Gastrulation in Mammals
• Cavities form above and below ICM,
which expand (as amnion and yolk sac,
respectively), leaving a flat 2-layered
plate of cells.
• Primitive streak forms as in birds and
reptiles to produce primary germ layers.
Figs 5.15 & 5.16 – Gastrulation in Mammals
Major Tissue Regions after Gastrulation
• Skin ectoderm
• Neural ectoderm
• Notochordal mesoderm
• Lateral mesoderm
• Endoderm
NEURAL TUBE FORMATION – Amphioxus
1. Folding up of tissue at junction of future skin
ectoderm and neural ectoderm areas; the two tissues
separate as this fold forms
2. Skin ectoderm grows over the top of neural ectoderm
3. Beneath “skin,” lateral margins of neural ectoderm
grow upward and together to form tube
4. Tube first closes at midpoint, progresses anteriorly
and posteriorly. Anterior end opens to surface as
neuropore, posterior end forms common opening
with blastopore (becomes anus) forming neurenteric
canal.
Fig 5.7 – Gastrulation and
Neurulation in Amphioxus
NEURAL TUBE FORMATION – Vertebrates

1. Formation of neural folds along margins of

skin-neural ectoderm
2. Mid-dorsal meeting of folds, simultaneous
with joining of skin ectoderm
3. During folding, high crests of tissue are
formed on either side = neural crest cells
Fig 5.16 – Gastrulation
and Neurulation in
Mammals
 MESODERM DEVELOPMENT
• Majority of body structures are mesodermal in origin.
• Notochordal Mesoderm rapidly rounds up and
separates from lateral mesoderm, forming a discrete
cylinder = notochord.
– Notochord is much reduced or obliterated in most adult
vertebrates, but forms the center around which vertebral
formation occurs.
• Lateral Mesoderm – Amphioxus
– Mesoderm forms paired series of segmentally arranged
blocks = somites.
– From their initiation, somites have a cavity inside = coelomic
cavity.
Fig 5.7 (b) – Notochordal and lateral mesoderm in Amphioxus
 MESODERM DEVELOPMENT
• Lateral Mesoderm – Vertebrates
• Initially there is no segmentation of mesoderm; instead forms as
a continuous sheet without a central cavity.
• Mesodermal differentiation occurs from dorsal midline outward
into 3 divisions, each extending the entire length of the body
trunk.
• Differentiation always occurs head-to-tail.
• The 3 divisions are:
1. Next to neural tube and notochord = Epimere (somites). Thicken and
subdivide on either side to form longitudinal rows of blocks. This is
the first indication of segmentation in vertebrate embryos.
Proliferation and differentiation occurs within somite forming:
• Sclerotome = portion surrounding notochord and neural tube
• Dermatome = outermost portion near skin ectoderm
• Myotome = middle portion between and ventral to sclerotome and dermatome
 MESODERM DEVELOPMENT
2. Lateral and ventral to somites is a relatively
small region of mesoderm, known as
intermediate mesoderm or Mesomere. This
may show segmentation similar to somites.
3. Beyond mesomere region, extending
ventrolaterally is a sheet of mesoderm known
as lateral plate mesoderm or Hypomere.
– Apart from cyclostomes, there is no segmentation
in this region. Coelomic cavity forms within lateral
plate mesoderm, dividing it into:
• Somatopleure = external mesoderm + ectoderm
• Splanchnopleure = internal mesoderm + endoderm
Figs 5.11 & 5.16
Mesoderm divisions
in Amphibians and
Mammals
 Organogenesis/Differentiation
• Once the mesoderm divisions are set up, then
ontogenetic development proceeds to
embryonic differentiation to adult body.
• What causes this differentiation?
• Induction = process by which developmental
fate of cells is determined
 Induction
• Classic Experiments of Spemann (1920s) – won Nobel Prize
• Took piece of presumptive neural plate from early gastrula stage,
transplanted to ventral region of another early gastrula  normal
development, prospective neural ectoderm becomes skin ectoderm.
• Similar experiment with late gastrula  transplanted neural
ectoderm becomes neural ectoderm regardless of transplant site.
• Transplant dorsal lip (prospective notochordal mesoderm) of early
gastrula to ventral region in another embryo  induces formation
of a secondary embryo involving both host and transplanted tissues.
• Conclusions:
– Some change took place between early and late gastrula that determined fate of
cells
– Dorsal lip responsible for inducing shift in direction of differentiation of host
tissue. Dorsal lip plays a central role in determining the craniocaudal axis of the
embryo = Primary Organizer.
Spemann Expt - Transplanted dorsal lip (prospective notochordal mesoderm) of
early gastrula to ventral region in another embryo  induces formation of a
secondary embryo involving both host and transplanted tissues.
 Primary Induction
• Definition = induction with primary
importance in determining cranio-caudal axis,
also the first of many inductive interactions.
• Two types of inductive interactions:
– Instructive = directs differentiation of cells along a
certain path
– Permissive = allows differentiation when given a
proper stimulus
 What is the Mechanism of Induction?
• Classic Triturus (Salamander) experiment …
• Dorsal Lip (Inducer) cultured for 24 hr across
nitrocellulose membrane with pores of specific
diameter from undifferentiated ectoderm → Ectoderm
differentiates to become neural tissues.
• If undifferentiated ectoderm cultured in the absence of
dorsal lip, it develops into unspecialized skin ectoderm.
• EM analysis of membrane after culture showed no
cellular processes between dorsal lip and ectoderm.
• Conclusion = a diffusable substance responsible for
induction.
 What is the Mechanism of Induction?
• Chemical nature of diffusable substance?
– Not known with certainty
• Purified active ingredients from various
inducers turn out to be proteins or
glycoproteins
• Also some evidence that changes in ratio of
bound/free ions within cells of early gastrula
may influence induction.
 What is the Mechanism of Induction?
• Other inductive interactions between cells can
result from …
– Cell-to-cell direct physical interactions – usually
between molecules located on the cell surface
– Cell-to-cell communication of signals through gap
junctions
• For induction via diffusable substances or
direct physical interactions, cell membrane
receptors on the induced cells are required
 SUMMARY
• The full developmental pathway is dependent
upon the genetic and biochemical capacities
of the induced cells + the full inductive
capabilities of the inducing tissue.
• Many inductive interactions occur throughout
development and influence gene expression
and the production of specific gene products
and cell migration, among other processes
• Precise mechanisms remain uncertain in most
cases.