The third week of

development

Dr kibe
Trilaminar germ disc

 
• Of particular significance during the third week is the
formation of the primitive streak on the surface of the
epiblast.
• The streak is seen as a narrow groove with slightly bulging
regions on either side.
• The cephalic end of the streak is known as the primitive node.
• The primitive node surrounds a small pit =the primitive pit
• The cells of the epiblast migrate towards and in the direction
of the primitive streak.
• When they arrive, they detach from the epiblast and sip
underneath it (invigination).
• Once the cells have arrived between the epiblast and
hypoblast, they spread laterally to form an intermediate cell
layer, the mesoderm germ layer in a process known as
gastrulation
• Some cells do not spread laterally but rather proceed and
displace the hypoblast and form the endoderm germ layer.
• The remaining epiblast cells becomes the ectoderm germ
layer
Notochord formation

• The cells (Prenotochordal cells) invaginating in the primitive pit move

straight forward in cephalic direction until they reach the prechordal
plate (buccopharyngeal membrane),
• where they form prechordal membrane. Prenotochordal cells
become intercalated in hypoblast so that 2 cell layers form
notochordal plate.
• The notochordal plate detaches from the endoderm and
retreats into the mesoderm forming a solid cord of cells →
definitive notochord.
• The notochord forms midline axis, which will serve as the
basis of the axial skeleton. See figures below
Prenotochordal cells invaginating in primitive pit move forward until they reach the buccopharyngeal membrane and
form the Prechordal mesoderm
 
Prenotochordal cells become intercalated in hypoblast so that 2 cell layers form notochordal plate
The notochordal plate detaches from the endoderm and retreats into the mesoderm forming a solid cord of cells →
definitive notocord
Establishment of the Body Axes
• Establishment of the body axes, anteroposterior,
dorsoventral, and left-right, takes place before and during
the period of gastrulation.
• The anteroposterior axis is signaled by cells at the anterior
(cranial) margin of the embryonic disc.
• This area, the anterior visceral endoderm (AVE), expresses
genes essential for head formation, including the
transcription factors OTX2, LIM1, and HESX1 and the
secreted factor cerberus.
• These genes establish the cranial end of the embryo before
gastrulation.
• The primitive streak itself is initiated and maintained by
expression of Nodal, a member of the transforming growth
factor β (TGF-β) family (Fig. 4.5).
• Once the streak is formed, a number of genes regulate
formation of dorsal and ventral mesoderm and head and tail
structures
• Another member of the TGF-β family, bone morphogenetic
protein-4 (BMP- 4) is secreted throughout the embryonic
disc (Fig. 4.5).
• In the presence of this protein and fibroblast growth factor
(FGF), mesoderm will be ventralized to contribute to kidneys
(intermediate mesoderm), blood, and body wall mesoderm
(lateral plate mesoderm)
• In fact, all mesoderm would be ventralized if the activity of
BMP-4 were not blocked by other genes expressed in the
node. For this reason, the node is the organizer.
• It was given that designation by Hans Spemann, who first
described this activity in the dorsal lip of the blastopore, a
structure analogous to the node, in Xenopus embryos.
• Thus, chordin (activated by the transcription factor
Goosecoid ), noggin, and follistatin antagonize the activity of
BMP-4.
• As a result, cranial mesoderm is dorsalized into notochord,
somites, and somitomeres
• Later, these three genes are expressed in the notochord and
are important in neural induction in the cranial region.
• As mentioned, Nodal is involved in initiating and maintaining
the primitive streak
• Similarly, HNF-3β maintains the node and later induces
regional specificity in the forebrain and midbrain areas.
Without HNF-3β, embryos fail to gastrulate properly and lack
forebrain and midbrain structures.
• As mentioned previously, Goosecoid activates inhibitors of
BMP-4 and contributes to regulation of head development.
• Overexpression or underexpression of this gene results in
severe malformations of the head region, including
duplications
• Regulation of dorsal mesoderm formation in mid and caudal
regions of the embryo is controlled by the Brachyury (T)
gene
• Thus, mesoderm formation in these regions depends on this
gene product, and its absence results in shortening of the
embryonic axis (caudal dysgenesis; )
• The degree of shortening depends upon the time at which
the protein becomes deficient
• Left-right sidedness, also established early in development, is
orchestrated by a cascade of genes.
• When the primitive streak appears, fibroblast growth factor
8 is secreted by cells in the node and primitive streak and
induces expression of Nodal but only on the left side of the
embryo
• Later, as the neural plate is induced, FGF-8 maintains Nodal
expression in the lateral plate mesoderm (Fig. 4.10), as well
as Lefty-2, and both of these genes
• upregulate PITX2, a transcription factor responsible for establishing
left sidedness
• Simultaneously, Lefty-1 is expressed on the left side of the floor plate
of the neural tube and may act as a barrier to prevent left-sided
signals from crossing over.
• Sonic hedgehog (SHH ) may also function in this role as well as
serving as a repressor for left sided gene expression on the right.
• The Brachyury(T) gene, another growth factor secreted by the
notochord, is also essential for expression of Nodal, Lefty-1, and Lefty-
2
• Genes regulating right-sided development are not as well defined,
although expression of the transcription factor NKX 3.2 is restricted to
the right lateral plate mesoderm and probably regulates effector
genes responsible for establishing the right side.
• Why the cascade is initiated on the left remains a mystery, but the
reason may involve cilia on cells in the node that beat to create a
gradient of FGF-8 toward the left.
• Indeed, abnormalities in cilia-related proteins result in laterality
defects in mice and some humans with these defects have abnormal
ciliary function
Fate Map Established During Gastrulation
• Regions of the epiblast that migrate and ingress through the
primitive streak have been mapped and their ultimate fates
determined
• For example, cells that ingress through the cranial region of
the node become notochord; those migrating at the lateral
edges of the node and from the cranial end of the streak
become paraxial mesoderm
• cells migrating through the midstreak region become intermediate
mesoderm; those migrating through the more caudal part of the
streak form lateral plate mesoderm;
• and cells migrating through the caudal-most part of the streak
contribute to extraembryonic mesoderm (the other source of this
tissue is the primitive yolk sac [hypoblast
Growth of the Embryonic Disc
• The embryonic disc, initially flat and almost round, gradually
becomes elongated, with a broad cephalic and a narrow
caudal end
• Expansion of the embryonic disc occurs mainly in the
cephalic region; the region of the primitive streak remains
more or less the same size
• Growth and elongation of the cephalic part of the disc are
caused by a continuous migration of cells from the primitive
streak region in a cephalic direction.
• Invagination of surface cells in the primitive streak and their
subsequent migration forward and laterally continues until
the end of the fourth week.
• At that stage, the primitive streak shows regressive changes,
rapidly shrinks, and soon disappears
• That the primitive streak at the caudal end of the disc
continues to supply new cells until the end of the fourth
week has an important bearing on development of the
embryo
• In the cephalic part, germ layers begin their specific
differentiation by the middle of the third week whereas in
the caudal part, differentiation begins by the end of the
fourth week.
• Thus gastrulation, or formation of the germ layers, continues
in caudal segments while cranial structures are
differentiating, causing the embryo to develop
cephalocaudally
C L I N I C A L C O R R E LAT E S
• Teratogenesis Associated With Gastrulation
• The beginning of the third week of development,
when gastrulation is initiated, is a highly sensitive
stage for teratogenic insult.
• At this time, fate maps can be made for various organ
systems, such as the eyes and brain anlage, and these
cell populations may be damaged by teratogens
• For example, high doses of alcohol at this stage kill cells in
the anterior midline of the germ disc, producing a deficiency
of the midline in craniofacial structures and resulting in
holoprosencephaly.
• In such a child, the forebrain is small, the two lateral
ventricles often merge into a single ventricle, and the eyes
are close together (hypotelorism).
• Because this stage is reached 2 weeks after fertilization, it is
approximately 4 weeks from the last menses.
• Therefore, the woman may not recognize she is
pregnant, having assumed that menstruation is late
and will begin shortly.
• Consequently, she may not take precautions she
would normally consider if she knew she was
pregnant.
• Gastrulation itself may be disrupted by genetic
abnormalities and toxic insults.
• In caudal dysgenesis (sirenomelia), insufficient
mesoderm is formed in the caudal-most region of the
embryo. Because this mesoderm contributes to
formation of the lower limbs, urogenital system
(intermediate mesoderm), and lumbosacral vertebrae,
abnormalities in these structures ensue
• Affected individuals exhibit a variable range of defects,
including hypoplasia and fusion of the lower limbs, vertebral
abnormalities, renal agenesis, imperforate anus, and
anomalies of the genital organs
• In humans, the condition is associated with maternal
diabetes and other causes.
• In mice, abnormalities of Brachyury (T), Wnt, and engrailed
genes produce a similar phenotype.
• Situs inversus is a condition in which transposition of the
viscera in the thorax and abdomen occurs.
• Despite this organ reversal, other structural abnormalities
occur only slightly more frequently in these individuals.
• Approximately 20% of patients with complete situs inversus
also have bronchiectasis and chronic sinusitis because of
abnormal cilia (Kartagener syndrome).
• Interestingly, cilia are normally present on the ventral surface
of the primitive node and may be involved in left-right
patterning during gastrulation.
• Other conditions of abnormal sidedness are known as
laterality sequences.
• Patients with these conditions do not have complete situs
inversus but appear to be predominantly bilaterally left sided
or right sided
• The spleen reflects the differences; those with left-sided
bilaterality have polysplenia, and those with right-sided
bilaterality have asplenia or hypoplastic spleen.
• Patients with laterality sequences also are likely to have
other malformations, especially heart defects.
Sirenomelia (caudal dysgenesis). Loss of mesoderm in the lumbosacral
region has resulted in fusion of the limb buds and other defects
Tumors Associated With Gastrulation

• Sometimes, remnants of the primitive streak persist in the

sacrococcygeal region.
• These clusters of pluripotent cells proliferate and form
tumors, known as sacrococcygeal teratomas, that commonly
contain tissues derived from all three germ layers
• This is the most common tumor in newborns, occurring with
a frequency of one in 37,000.
• These tumors may also arise from primordial germ cells
(PGCs) that fail to migrate to the gonadal ridge
Sacrococcygeal teratoma resulting from remnants of the primitive streak.
These tumors may become malignant and are most common in females
• By the beginning of the third week, the trophoblast is characterized
by primary villi that consist of a cytotrophoblastic core covered by a
syncytial layer
• During further development, mesodermal cells penetrate the core of
primary villi and grow toward the decidua. The newly formed
structure is known as a secondary villus (Fig. 4.15).
• By the end of the third week, mesodermal cells in the core
of the villus begin to differentiate into blood cells and
small blood vessels, forming the villous capillary system
• The villus is now known as a tertiary villus or definitive
placental villus.
• Capillaries in tertiary villi make contact with capillaries
developing in mesoderm of the chorionic plate and in the
connecting stalk
• These vessels, in turn, establish contact with the
intraembryonic circulatory system, connecting the
placenta and the embryo.
• Hence, when the heart begins to beat in the fourth
week of development, the villous system is ready to
supply the embryo proper with essential nutrients and
oxygen.
• Meanwhile, cytotrophoblastic cells in the villi penetrate
progressively into the overlying syncytium until they reach
the maternal endometrium.
• Here they establish contact with similar extensions of
neighboring villous stems, forming a thin outer
cytotrophoblast shell
• This shell gradually surrounds the trophoblast entirely and
attaches the chorionic sac firmly to the maternal
endometrial tissue
• Villi that extend from the chorionic plate to the
decidua basalis (decidual plate: the part of the
endometrium where the placenta will form are called
stem or anchoring villi.
• Those that branch from the sides of stem villi are free
(terminal) villi, through which exchange of nutrients
and other factors will occur
• The chorionic cavity, meanwhile, becomes larger, and
by the 19th or 20th day, the embryo is attached to its
trophoblastic shell by a narrow connecting stalk
• The connecting stalk later develops into the umbilical
cord, which forms the connection between placenta
and embryo.
Problem solving
• A 22-year-old woman consumes large quantities of alcohol at
a party and loses consciousness; 3 weeks later she misses her
second consecutive period. A pregnancy test is positive.
Should she be concerned about the effects of her binge-
drinking episode on her baby?
• An ultrasound scan detects a large mass near the sacrum of
a 28-week female fetus. What might the origin of such a
mass be, and what type of tissue might it contain?
• On ultrasound examination, it was determined that a fetus had well-
developed facial and thoracic regions, but caudal structures were abnormal.
Kidneys were absent, lumbar and sacral vertebrae were missing, and the
hindlimbs were fused. What process may have been disturbed to cause such
defects?
• A child has polysplenia and abnormal positioning of the heart. How might
these two abnormalities be linked developmentally, and when would they
have originated?
Should you be concerned that other defects might be present?
What genes might have caused this event, and when during embryogenesis
would it have been initiated?