CFB L1-4: General Embryology

General Embryology I
Primordial germ cells
Gametogenesis
- Meiosis
- Oogenesis
- Spermatogenesis
- Fertilization
- Cleavage
- Blastocyst formation and implantation
Timeline of human development
Total fetal development takes 38 weeks from the time of fertilization
Embryonic period — up to week 8
Fetal period — after the first 8 weeks
Primordial germ cells
Development begins with fertilization
Fertilization — fusion of haploid gametes, oocyte and sperm
Primary function of gametes is to transmit genetic information
Gametes are derived from primordial germ cells
Primordial germ cells (PGCs) are formed during the 2 nd week
PGCs arise from the epiblast
- Epiblast is the region of the germ layers that gives rise to all the layers of the developing organism
Move to the wall of the yolk sac (~3rd week)
- PGCs arise from the region of the epiblast that will move into the extraembryonic tissue into the wall of the yolk sac
During the 4th week, PGCs migrate from the yolk sac back towards the genital ridge
- Genital ridge is the precursor for gonads
PGCs migrate along the gut tube and the dorsal mesentery
- Migrate along the hindgut
PGCs invade the developing gonad by the 6th week
PGCs proliferate during migration and for several days after they enter the gonad
The sex of the embryo is determined genetically after fertilization, however, the gonads do not acquire male or female
characteristics until the 7th week
Upon arrival of PGCs, the epithelium of the genital ridge proliferates into the underlying mesenchyme to form the primitive sex
cords
At this stage, it’s impossible to differentiate between male and female gonads
- The gonad is known as indifferent gonad (before it has differentiated into male/female gonads)
Genetically male embryos have an XY chromosome complex
- The Y chromosome carries the SRY gene, the testis-determining factor
- Under the influence of SRY, the primitive sex cords differentiate into medullary/testis cords  testis
Genetically female embryos have an XX sex chromosome complex
- In the absence of SRY function, the primitive sex cords degenerate
- The surface epithelium proliferate to give rise to cortical cords  ovary
In the absence of a Y chromosome, we default to female due to the absence of the SRY gene
Meiosis
A unique division process, which occurs exclusively in germ cells
2 major functions;
- Meiotic recombination
- Reduction of the number of chromosomes to a haploid complement
Key events
- 1. One round of (pre-meiotic) DNA replication
- 2. A unique prophase I stage of meiosis for genetic exchange
- 3. Two rounds of chromosome segregation, meiosis I and meiosis II (thus the number of chromosomes in reduced to a
haploid complement
The initiation of meiosis marks the transition from PGC to gamete
Mitosis
- Cell division giving rise to 2 daughter cells that are genetically identical
- Each cell receives the complete complement of 46 chromosomes
Meiosis
- Consists of 2 successive meiotic divisions – meiosis I and meiosis II
 - Reduction of the number of chromosomes to a haploid number of 23
Prophase I of meiosis
Meiotic recombination: Genetic exchange between maternal and paternal alleles
Stages of prophase I:
- 1. Leptotene: condensation of chromosomes
- 2. Zygotene: pairing/synapsis of homologous chromosomes
- 3. Pachytene: crossing over/meiotic recombination
- 4. Diplotene: synapsed chromosomes begin to separate
- “Let Zebras Play Dead”
Production of gametes in female and male
Females:
- Primary oocytes undergoes meiosis I, giving rise to a secondary
oocyte and a polar body
o First maturation division gives rise to asymmetrical
secondary oocyte and polar body
o One cell has most of the cytoplasm and the other has very
little
- Meiosis II produces a mature oocyte and a second polar body
o Again, asymmetrical cell division
- Polar bodies ensure we give rise to only ONE single mature
gamete
Males:
- Primary spermatocyte gives rise to TWO spermatocytes, which give rise to FOUR spermatids
- Thus males give rise to FOUR mature gametes!
- Maturation division in males is symmetrical
Germ cell development in the male genital ridge
Germ cells are committed to spermatogenesis
Sertoli cells have differentiated and testis cords have formed
Germ cells enter mitotic arrest, forming spermatogonia
Mitosis is not resumed until after birth
The first meiotic division is initiated at puberty
The second meiotic division is completed in adulthood and occurs through life
Germ cell development in the female genital ridge
Germ cells are committed to oogenesis
All germ cells synchronously enter meiotic prophase and become oocytes
Undergo the early stages of meiosis (meiotic recombination)
Oocytes arrest at diplotene prophase I at the time of birth
First meiotic division is completed at puberty
The first meiotic prophase occurs as a fetus, then you remain in resting stage (diplotene) as a newborn, and complete meiosis I at
puberty
The second meiotic division only occurs upon fertilization as an adult
Oogenesis
The process by which oogonia differentiate into mature oocytes
Once PGCs arrive in the gonad of a genetic female, they differentiate into oogonia
Oogonia undergo a number of mitotic divisions during fetal life
Some oogonia arrest their division at prophase of meiosis I to form primary oocytes
- Occurs around the 7th month of gestation
- At this stage, the cells are surrounded by follicular epithelial cells
Primary oocytes become individually surrounded by a layer of follicular epithelial cells (primary follicular cells)
- They are known as primordial follicles
At puberty, a selected number of primary oocytes grow and the follicular cells proliferate to produce a stratified epithelium of
granulosa cells — they are the primary follicles
The primary oocytes become surrounded by a layer of glycoproteins — zona pellucida
The growing follicles are passing through 3 stages:
- Primary/pre-antral
- Secondary/antral (central cavity is the antrum)
- Pre-ovulatory
As development proceeds, secondary follicle develops a fluid filled space, the antrum
Over time, the antrum enlarges with granulosa cells surrounding the oocyte
This layer of cells is known as corona radiata (forma crown of cells around the oocyte)

Upon lutenizing hormone stimulation, meiosis I is completed

- 1st polar body
The oocyte enters meiosis II and arrests in metaphase
Ovulation takes place
Meiosis II is completed only upon fertilization
Spermatogenesis
The process by which spermatogonia are transformed into spermatozoa
Begins at puberty and continues into old age
Meiosis and sperm maturation begin at puberty in the seminiferous tubules
Self-renewing, mitotic, spermatogonial stem cell population (continuous process)
Primary spermatocytes  meiosis I  secondary spermatocytes  meiosis II  spermatids
Spermiogenesis
Transformation of haploid spermatids into sperm
- Spermatids change shape to mature sperm
- Last phase of male sperm maturation
Acrosome formation
- Acrosome contains enzymatic proteins that allow sperm to
enter the oocyte
- Most of the nucleus is here
Condensation of the nucleus
Loss of cytoplasm
- Via digestion by supporting cells found in the testes
Middle piece and tail formation
Key differences in male and female gametogenesis
Oogenesis
- Finite population of oocytes established at birth
- 1 gamete produced by each meiosis event
- Meiosis begins during fetal development, but is not resumed until puberty
Spermatogenesis
- Continuously dividing stem cell population
- 4 gametes produced per meiotic division
- Meiosis initiated at puberty
Fertilization
Fertilization is the fusion of haploid gametes, egg and sperm
Results in the formation of the diploid embryo
The beginning of embryogenesis
Fertilization occurs in the ampullary region of the
fallopian tube
- The most distal region of the tube
Initial movement of sperm into female genital tract is due to
female genital tract contractions, the motility of the sperm
is only used in the last few steps
Ovulated oocyte
Surrounded by cells of the corona radiata
Surrounded by a glycoprotein coat, zona pellucida
Completed the first meiotic division
Extruded one set of chromosomes into the first polar body (not involved in fertilization)
Arrest at metaphase of 2nd meiosis
If no fertilization occurs, then eventually the ovulated oocyte will degenerate
Key features of fertilization
1. Capacitation of the sperm
- Conditioning of the sperm in the female reproductive tract
- Ejaculated sperm must stay in female reproductive tract for at least 7 hours before it’s ready to fertilize the egg
- Remove surface glycoproteins, especially from acrosome, allowing contact of sperm and oocyte
2. Contact and recognition
- Binding to species-specific receptors on the zona pellucida (ZP3)
3. Acrosome reaction
- Release of proteolytic enzymes to digest the zona
pellucida
4. Sperm fusion with the egg membrane
- Fusion of membrane stimulates cortical granule reaction
- Cortical granule reaction in the egg — blocks polyspermy
(prevents other sperm from entering the egg)
5. Egg activation
- Completion of meiosis (second meiotic division)
6. Fusion of the genetic material
- Formation of the diploid embryo (zygote)
Cortical granule reaction
Fusion of the plasma membrane of sperm and oocyte
Prevents other sperm from entering the egg
Sperm acrosome releases proteolytic enzymes to digest zona pellucida so the plasma membranes can fuse
Plasma membrane of oocyte is lined with cortical granules
Fusion of plasma membranes causes granules to be released into the space between the plasma membrane of the oocyte and zona
pellucida  separation, preventing other sperm from binding
Zygote
Fertilization produces a zygote
Two pronuclei (male and female) fuse to give fertilization
Most cytoplasm of the zygote comes from the female oocyte and not the male sperm
Cleavage
Zygote undergoes a series of mitotic divisions — cells known as
blastomeres
- Referring to mitosis now, not meiosis
Embryonic genome activation
Initiation of gene transcription from the embryonic genome that
will regulate subsequent development
1. Degradation of material mRNA
2. Minor ZGA — zygotic genome activation
- At the one-cell stage
3. Major ZGA
- At the 2- and 4-cell stages
First phase of division = zygotic genome activation
Prior to fertilization, survival of oocyte to one-cell stage is
dependent on accumulation of maternal mRNA during
oogenesis
In the fertilization 2- and 4-cell stages, maternal mRNA
decreases so we get a burst of zygote
transcription/translation, producing mRNA that is essential to
sustain the developing embryo
Compaction
Around the 8-cell stage the embryo goes through the process of compaction (blastomeres maximize their contact with each other)
- Morula will go through compaction and the blastomeres begin to flatten against each other
Cellular events associated with compaction:
- Increase in tight junction formation
- Production of E-cadherin (cell-cell adhesion molecule)
- Formation of gap junctions
 - Shape change in outer blastomeres
- Inner/outer polarity in cells (predicts inner/outer polarity of the embryo)
Compaction increases surface contact of embryo
Compaction takes place 3 days after fertilization in the uterus
- Then the morula continues forward towards the uterus
Blastocyst formation and implantation
Blastomeres remaining in contact with the outside give rise to the
trophectoderm
Blastomeres inside the embryo give rise to the embryo proper/inner cell
mass (ICM)
Blastocele (cavity) forms progressively through build up of fluid inside the morula
Have asymmetrical cell mass that begins to grow on one side of the blastocyst formed by the inner cell mass
Segregation of trophectoderm and ICM lineages depends on CDX2 and OCT4 expression
If you express the CDX2 genes then you will give rise to the trophectoderm
But if you express OCT4 then you will give rise to inner cell mass cells
Unique properties of the inner cell mass
Source of embryonic stem cells (ESCs)
ESCs can self-renew (unlimited division)
ESCs are pluripotent (OCT4)
- Can express many factors, including OCT4
Can give rise to cell types from all three primary germ layers
The ICM is the only cell layer that contributes to the growing embryo; no other cells do
Hatching from the zona pellucida
When the blastocyst reaches the uterus, it must “hatch” from the zona pellucida in order to implant into the uterine wall
- Blastocyst starts to move out of zona pellucida
- If embryo doesn’t hatch, it can’t implant
Trophoblast cells secrete protease to digest the zona pellucida
Hatching occurs around 4.5-5 days after fertilization
Next, the embryo implants in the uterus wall
Implantation occurs at 5.5-6 days
Blastocyst implantation
The hypoblast segregate from the inner cell mass
The rest of the inner cell mass form the embryonic epiblast
The amniotic cavity appears in the epiblast
 Bilaminar embryonic disk
Segregation of epiblast and hypoblast from the ICM depends on GATA6 and NANOG expression
We have another transcription factor that gives rise to the hypoblast and epiblast
Note how different TFs appear at different stages and give rise to different structures of the embryo
Recall, COX2  trophoectoderm while OCT4  ICM
ICM + GATA6  hypoblast
ICM + NANOG  epiblast
Derivatives of the epiblast and hypoblast
Epiblast gives rise to almost all structures of the embryo (endoderm, mesoderm, ectoderm, notochordal process, etc)
Hypoblast underlies the embryo and gives rise to the yolk sac and doesn’t contribute to the embryo proper
- Only forms the extraembryonic mesoderm
7.5-day human blastocyst

Process of implantation
- Epiblast is in blue, hypoblast is in yellow
 - Epiblast has another cavity that is forming the amniotic cavity
- Cytorophoblast and syntrophoblasts (the green cells)
o Derived from trophoblasts
o Cells that are digestive and helping with implantation into the uterine wall
H&E staining of epiblast/hypoblast at 7.5 days
- Can see trophoblast starting to invade the epiblast
9-day human blastocyst

12-day human blastocyst

13-day human blastocyst

Blastocyst is completely surrounded by endometrium of the mother here

General embryology II
Gastrulation and germ layers formation
Derivatives of the ectoderm
- Neural induction
- Neurulation
- Neural crest cell derivatives
- Cranial placodes development
Timeline of human development
Recall, total fetal development takes 38 weeks from the time of fertilization
- Embryonic period — up to week 8
- Fetal period — after the first 8 weeks
Second half of the embryonic period is from weeks 3-8
Once we form a blastocyst, it will give rise to both germ cells and somatic cells
Germ cells — form gametes; reproduction
Somatic cells — form all body (soma) cells
Somatic cells are the cells forming the body of an organism (tissue and organs), as opposed to the germline, which form the gametes
(spermatozoa and oocyte)
Somatic cells are entirely derived from the three embryonic germ layers — ectoderm, mesoderm, and endoderm — which are
established during gastrulation
Gastrulation and germ layer formation
If you fail to go through gastrulation, you fail to form a proper embryo
- Gastrulation is the most important event in your life!
A process characterized by profound cell movements that convert a sheet of cells
into a multi-layered organism
One of the consequences of this reorganization is that groups of cells are
brought in close contact to undergo inductive interactions
The major outcome is the formation of the three germ layers of the embryo
15 days
At 15 days (end of the 2nd week of gestation), the implanting blastocyst begins to
undergo gastrulation
- Blastocyst has 2 layers — epiblast and hypoblast
First indication of gastrulation is an accumulation of cells and formation of a
groove, the primitive streak, at the caudal end
- Primitive streak forms down the epiblast center, and creates a primitive node
- Cells of the epiblast move into this node, into the cavity of the embryo
16 days
The primitive streak is initially formed in the caudal part of the
embryo and moves rostrally
The most anterior part of the groove is the primitive node
At the primitive streak, the cells of the epiblast are moving into the
cavity between the epiblast and hypoblast layers
- Moving deep into the groove
Cells that move rostrally/anteriorly from the primitive node give rise to
the notochord
Cells move towards the lateral edge form the primitive streak
- The first wave of cells will first move down from the epiblast deep
into hypoblast, replace the cells of the hypoblast, and push the
cells of the hypoblast laterally
o As those cell are not part of the embryo, these new cells
will become the endoderm
- The second wave of cells come from the epiblast, move
between the hypoblast and epiblast, and give rise to the
mesoderm germ layer
Key events associated with gastrulation
Epiblast cells forming the primitive streak migrate in an anterior position to form the node
First cells passing through the primitive streak displace the cells of the hypoblast to give rise to the embryonic/definitive endoderm
Cells at the node move anteriorly to form the head mesoderm and the notochord as the primitive streak regresses
Cells moving through the primitive streak between the epiblast and the endoderm will form
the embryonic mesoderm
Epiblast cells that remain at the surface will form the embryonic ectoderm
The oropharyngeal membrane at the cranial end will break down to form the stomedeum (mouth opening)
The cloacal membrane at the caudal end will break down to form the opening of the anus
First, primitive streak forms at the posterior of embryo — a groove for cells of epiblast to move deeper into embryo
These cells intercalate with cells of the hypoblast and push these cells laterally so these hypoblast cells aren’t part of the embryo
The cells moved deep then give rise to the endoderm
Second wave of cells that pass through the primitive streak and intercalate with the newly formed endoderm become the mesoderm
Cells that migrate through the node and move anteriorly, give rise to the notochord
What remains on the top is the ecotderm
There are two regions where mesoderm doesn’t come between ectoderm and endoderm — at the posterior and anterior regions of
the embryo
- At the oropharyngeal membrane there’s no mesoderm between the ecto/endoderm
o Oropharyngeal membrane becomes the opening of the mouth
- The same occurs at the cloacal membrane, which forms the opening of the anus
Derivatives of the ectoderm
Ectoderm is the most superficial layer
Epidermis
CNS
Cranial placodes (sense organs)
Neural crest
- PNS
- Pigment cells
- Craniofacial skeleton
- Odontoblasts
- Adrenal medulla
Neural induction
The process by which the neural plate is generated
Inductive interaction between the chordamesoderm/notochord and the ectoderm
Gastrulation  neural induction  neurulation
The notochord is the structure that sends the signal for the development of the neural plate
Neurulation — newly formed neural plate induces formation of the CNS (brain and spinal cord)
Molecular basis of neural induction
The secreted molecule, bone morphogenetic protein 4 (BMP4), expressed throughout the ectoderm, regulates the decision between
epidermal and neural fate
- BMPs are produce by one cell, but signal to neighboring cells that they will become epidermis/skin tissue
Ectoderm produces BMP
- BMP uninterrupted  epidermis
- Blocked BMP  neural tissue
Inhibition of BMP4 signaling in the ectoderm is required to convert the ectoderm into neural tissue (mediated by the dorsal
mesoderm)
Noggin, Chordin, and Follistatin are secreted by the dorsal mesoderm/notochord
- They bind BMP4 molecules, preventing BMP4 interaction with its receptor
- They act as BMP antagonists
- Blocking BMP4 binging prevents formation of the epidermis, thus promoting formation of neural tissues
Regionalization of the neural plate
The nervous tissue induced by BMP4 inhibition is anterior in character (forebrain)
Another set of factors interact with the BMP antagonists  anterior neural tissue formation and further differentiation of the
nervous system from anterior to posterior character (aka activation then transformation)
1. Activation — BMP4 is blocked to give rise to neural tissue instead of epidermal tissue
 - Mediated by Noggin, Chordin, Follistatin
2. Transformation — neural tissue is converted from anterior to posterior character
- Mediated by FGF, retinoic acid, Wnt-3a
Neurulation
The process by which the neural plate folds into the neural tube
Neurulation results in the segregation of the three ectodermal derivatives
- Neural tube, neural crest, and epidermis
Stages of neural tube formation:
- Shaping  folding  elevation  convergence  closure
- Neurulation causes the neural tube to start to thicken
o Thickens through shaping
- The neural tube then folds, and the edges of the neural plates
start to rise
- Then the neural plate edges fuse, and the epidermis covers
them
- The cells at the most dorsal aspect of the neural tube are the
neural crest cells
Neural tube closure is initiated at the level of the future midbrain and
zips up in both directions
19 days — folding begins; neural plate starts to close (closes from the center of the embryo)
20 days — folding starts in the middle of the neural plate
22 day — neural tube is closed in the middle, but open on the cranial and caudal sides
- The tube zips up/down from the middle
23 days — neural tube is almost entirely closed, except the cranial (anterior) and caudal (posterior) neuropores
- The last regions to close
Neural tube closure defects
Among the most common congenital anomalies
Anencephaly — failure to close the anterior part of the neural tube
- Lack of skull bone and degeneration of brain structure
Craniorachischisis — open brain and spinal cord
Spina bifida — failure to close the neural tube in the caudal region
- Severity depends on how much of the spinal cord is exposed through the vertebral arches and other superficial structures
(spina bifida cystica)
- Major types:
o Occulta — arch is slightly open but doesn’t have that many consequences
o Meningocele — meninges are exposed
o Myelomeningocele — meninges are spinal cord are exposed
o Cystica — the most severe form; the meninges have a cyst forming in them
Neural crest development
Neural crest cells derive from ectoderm in the region between the
neural plate and future epidermis, at the dorsal neural
Migratory behavior is associated with changes in cell adhesion
properties
Epithelial-to-mesenchymal transition
- More loose organization of the cells
Neural crest cells migrate away from the dorsal part of the neural tube and go throughout the body
Functional domains of the neural crest
There are 5 different regions of neural crest cells, based on migration
Cranial neural crest — hindbrain and midbrain
- Cartilage, bone, cranial ganglia, and connective tissue of the face
- Cranial neural crest cells (derived from the hindbrain) migrate dorsolaterally and contribute to the craniofacial mesenchyme
that differentiates into cartilage, bone, cranial ganglia (neurons and glia) and connective tissues of the face
- Migration of the cranial neural crest from the hindbrain
o Migrate into the head region and form the pharyngeal branchial arches and give rise to specific elements of the
head
o Pharyngeal arches origin of craniofacial skeletal elements derived from the neural crest
 Neural crest cells in branchial arch #1 contribute to the mandible, zygomatic complex, and maxilla, and
also the malleus and incus
  Neural crest cells in branchial arch #2 contribute to the hyoid and stapes
 The branchial arch that is in the most anterior region, that populates the frontal neo-prominence, gives
rise to the frontal bone
Cardiac neural crest — septum of the heart
- Septum of the outflow tract of the heart and wall of the large arteries
- The cardiac neural crest cells can give rise to the entire musculo-connective tissue of the large arteries as they arise from
the heart and to the septum which separates the pulmonary artery from the aorta
- The septum that separates the truncus arteriosus into the pulmonary artery and aorta is derived from the cardiac neural
crest
Vagal neural crest — somite #7
- Parasympathetic enteric ganglia of the gut
Trunkal neural crest
- Melanocytes, DRG, sympathetic ganglia, and adrenal medulla
Sacral neural crest
- Parasympathetic enteric ganglia of the gut
The neural crest provides innervation to the entire digestive system
- Vagal neural crest cells contribute to the anterior part of the digestive tract (foregut and midgut) and the sacral neural crest
cells contribute to the posterior part of the digestive tract (hindgut)
Neurocristopathies
Group of diseases associated with abnormal development of the neural crest
Result of defects in neural crest cells themselves but also of defects in the environment through which they migrate
Defects affect only a single derivative of the neural crest, whereas other defects result in a wide array clinical manifestations
DiGeorge Syndrome
Rare congenital disease (1:4,000 live births)
Caused by a large deletion in chromosome 22 (Tbx1)
Commonly include heart defects and craniofacial malformations
- Shortening of the lower jaw
- Incomplete separation of oxygenated vs deoxygenated blood in the heart (septum does not form properly in the heart)
Hirschsprung’s disease
Rare congenital disease (1:5,000 live births)
Mutations in Sox10 and Endothelin Receptor B
Aganglionic megacolon (absence of enteric neurons in the colon)
- Improper innervation of the distal part of the colon)
Often associated with pigmentation defects and deafness
Treacher Collins Syndrome
Autosomal dominant craniofacial disorder (1:50,000 live births)
Mutations in the TCOF1 gene, which encodes the nuclear protein Treacle1
Numerous development anomalies, including small jaws, cleft palate, and middle/external ear defects
- Defects stem from neural crest cells that contribute to branchial arch 1 and 2, so they affect the maxilla, mandible, and
zygomatic complex (causes slanting of the eyes)
- Due to lack of TCOF1 gene, there is a tendency of less survival of neural crest cells
Cranial placodes development
Neural tube is almost closed at 25 days
At 28 days, the branchial arches are colonized by neural crest cells
- Lens placode  lens (vision)
- Otic placode  inner ear (hearing/balance)
- Olfactory placode  oral epithelium (smell)
- Together known as cranial placodes
Cranial placodes — thickening of the cephalic embryonic ectoderm
Cranial placodes contribute to the sense organs (vision, hearing, and olfaction)
The thickening can either occur through invagination or they can form vesicles and separate and then give rise to the sense organs
Lens placode
At the level of the diencephalon, we get bulges that protrude for the eye
- At the wall of the forebrain, we get formation of optic vesicles where the future eye will form
Bulges dictate the region to thicken and create the lens placode
- The optic vesicles come in contact with an epithelium and their interaction will make the epithelium start to thicken and
give rise to the lens placode
- The lens placode will invaginate and give rise to the lens vesicle for vision
Otic placode
In the ectoderm, we get thickening of the otic placode, which invaginates to give rise to otic vesicles and morphogenic processes
that give rise to inner ear structures (e.g. cochlea, semi-circular canals)
Development of the adenohypophysis (anterior pituitary)
Cranial placodes contribute/give rise to the adenohypophysis (anterior pituitary)
Anterior pituitary gives rise to hormone producing cells of the pituitary
The placode starts in the mouth in the oral ectoderm, and gives rise to Rathke’s pouch
Invagination in combination with the neurohypophysis gives rise to the anterior pituitary
Cranial placodes also contribute to cranial ganglia
Instead of invagination, some placodes will undergo
delamination, which produces trigeminal and epibranchial
placodes
Trigeminal placodes  CN V  ophthalmic and
maxillomandibular lobes
Epibranchial placodes  geniculate, petrosal, and nodose
ganglia

General embryology III

Derivatives of the mesoderm
- Regionalization of the mesoderm
- Development of the mesoderm layers
- Somites
- Axial skeleton
- Limb development
Derivatives of the endoderm
- Regionalization of the gut tube
- Liver and pancreas development
- Respiratory system
Derivatives of the mesoderm
Mesoderm is sandwiched between the ectoderm and endoderm
Notochord, head muscle, circulatory system, heart, kidneys, ovaries and testes (urogenital system), axial and limb skeleton, axial and
limb muscles, body wall, connective tissue of the skin, spleen
Regionalization of the mesoderm
During gastrulation specific regions of the epiblast migrate through different regions of the primitive streak to form the mesoderm
Fate mapping analyses indicate that mesodermal tissue regionalization depends on the position and the timing of cell passage
through the node and the primitive streak in an anterior to posterior direction
Depending on the position and timing, you get different types of mesoderm from the primitive streak
From anterior to posterior of the primitive streak:
- Notochord (cells migrate through the node)
- Paraxial mesoderm
- Intermediate mesoderm
- Lateral plate mesoderm
- Extraembryonic mesoderm
Subdivisions of the mesoderm at the end of neurulation:

-
Each part of the mesoderm gives rise to…
- Chordamesoderm (axial mesoderm)
o Notochord
- Paraxial mesoderm
o Head mesenchyme
 o Somites
 Sclerotome (cartilage, bone)
 Myotome (skeletal muscle)
 Dermatome (dermis)
 Syndetome (tendons)
- Intermediate mesoderm
o Kidney
o Gonads
- Lateral plate mesoderm
o Splanchnic (visceral) mesoderm (circulatory system)
o Somatic (parietal) mesoderm (body wall)
Development of the mesoderm layer
During neurulation, the neural plate starts to fold and close, and on each side there’s block of cells that forms a large mass which
forms the paraxial mesoderm
The lateral mesoderm (lateral to the paraxial mesoderm) remains quite thin, but you can start to see a cavity that forms, which will
be essential in defining the two layers of the lateral plate mesoderm: parietal and visceral
- As you continue with gastrulation, the parietal and visceral layers bend around until they fuse together
In between these two mesoderm layers you have the proliferation of the intra-embryonic mesoderm
Derivatives of the lateral plate mesoderm
Dorsal components for the somatic (parietal) mesoderm:
- Body wall, lining the outside of the peritoneal, pleural, and pericardial cavities
- Mesodermal components of the limbs (except muscles)
- Comes in close contact with the ectoderm
Ventral component forms the splanchnic (visceral) mesoderm:
- Covering the abdominal organs, lung, and heart
- Circulatory system (heart, blood, and blood vessels)
- Muscle and connective tissue of digestive and respiratory tracts
- Comes in close contact with the endoderm
Coelom — space between the two layers will form the body cavity
- Abdominal (peritoneal)
- Thoracic (pericardial and pleural)
Somatopleure = somatic (parietal) mesoderm + ectoderm
- Forms the ventral and lateral body wall
- Forms the membranes that line the peritoneal, pleural, and pericardial cavities
Splanchnopleure = splanchnic (visceral) mesoderm + endoderm
- Forms the wall of the gut and the thin serous membrane around each organ
Derivatives of the intermediate mesoderm
Intermediate mesoderm generates the urogenital system
Their development is interconnected anatomically
Urinary system
- Mesonephros and mesonephric duct (embryonic kidney)
- Metanephros and metanephric duct (adult kidney)
Genital system
- Gonads (testis/ovary)
- Genital tract
Somites development
Paraxial mesoderm cells become loosely organized into aggregation of cells called somitomeres
The somitomeres eventually separate from the presomitic paraxial mesoderm to form individual somites
Somites appear first in the anterior portion of the trunk, and new somites "bud off” form the unsegmented presomitic (PSM) at
regular intervals
Somite periodicity
Somites appear with a specific periodicity
The periodicity is a highly regulated process (species specific) which depends on a segmentation clock established by the cyclic
expression of genes of the Notch and Wnt signaling pathways
The number of somites is usually a good indicator of the age of an embryo
Somite differentiation
Four major components to the somite:
- 1. Sclerotome (cartilage, bones)
 - 2. Myotome (skeletal muscle)
- 3. Dermatome (dermis)
- 4. Syndetome (tendons)
When the somite is first separated from the presomitic mesoderm, cells are not committed to a specific lineage
As the somite matures, its various regions become progressively restricted to form only certain cell types
Somites are initially naive, without specificity, but begin to receive input about their fate and segregate
- Cells of the sclerotome begin to segregate into other cells such as the
dermamyotome (dermatome + myotome) and another
component that segregates ventrally to form the future
sclerotome
- Further delamination and migration around the notochord gives rise to
the sclerotome (vertebrae)
- The primaxial dermamyotome (muscles of the back) is the closest
to the neural tube
- The abaxial dermamyotome (muscles of the body wall and limb) is the
furthest from the neural tube
- Sclerotome is the most ventral
- Dermatome is most dorsal
The syndetome is the last component to form
- It forms at the boundary between the sclerotome and the myotome
- The syndetome gives rise to tendon progenitors
What determines cell fate within the somite?
- Location, location, location!
- The fate of somites is determined by surrounding cells
- Notochord secretes SHH (sonic hedgehog), which defines the most ventral portion of
the somite, and tells the cells to become sclerotome
- Lateral plate mesoderm secretes BMP4 and FGF5, which define the tissue further from the neural tube, and give rise to the
abaxial muscle
- Epidermis secretes Wnt, which contributes to formation of the dorsal part of the somite
- Nt3,Wnt1, and Wnt3a come from the dorsal part of the neural tube and contribute to the primaxial muscles

-
Axial skeleton
Development of the skull
- The skull can be divided into 2 parts — neurocranium and viscerocranium
- Neurocranium forms the protective case around the brain (paraxial mesoderm)
- Viscerocranium forms the skeleton of the face (neural crest)
- Intramembranous ossification — flat bones of the skull vault
- Endochondral ossification — bones of the base of the skull
- At birth, the flat bones of the skull are separated from each other by sutures (narrow seams of connective tissue), which are
derived from two sources:
o Neural crest cells (sagittal suture)
o Paraxial mesoderm (coronal suture)
- Fontanelles — where more than two bones meet, and sutures are wider
o Remain soft tissue — especially important for birthing
- Sutures and fontanelles remain membranous for a considerable time after birth
o Bones of the vault continue to grow after birth
o Allows for brain growth
 o Anterior fontanelle closes by 18 months of age
o Posterior fontanelle closes by 1 to 2 months of age
- Important to maintain flexibility to allow for brain growth
- Closure can have significant results on the development of the head
- Craniosynostosis — premature closure of one or more sutures of the skull
Vertebral column
- Vertebrae are derived from the sclerotome portions of the
somites
- The sclerotome portion of each somite undergoes a
process known as resegmentation
- Each vertebra is formed from the fusion of the caudal half of
one somite and the cranial half of its neighbor
o Each sclerotome contributes to two different
vertebrae
Derivative of the notochord
- The notochord will become the nucleus pulposus
(chondrocyte-like cells)
- It’s surrounded by the circular fibers of the annulus
fibrosus
- These two structure for the intervertebral disc
o Nucleus pulposus is the central region of the intervertebral disc
Limb development
Outgrowth of the embryonic body wall, consisting of mesenchyme derived from the somites and the lateral plate mesoderm
These precursors accumulate and proliferate under the epidermis to creates the limb bud
The mesenchyme of the limb bud is derived from the mesoderm
- Somites (muscle precursors)
- Somatic lateral plate (skeletal precursors)
Start to develop limbs at weeks 3-5
Get visibility of some individualized digits by week 8
The apical ectodermal ridge (AER) is an important organizing center in the limb bud
The zone immediately adjacent to the AER is the progress zone (PZ)
Interaction between the AER and the PZ is critical to limb development
- Signal comes from the AER, and stimulates the PZ and differentiation of cells
Proximal-distal axis of the limbs

The PZ participates in the establishment from the proximal-distal axis of the limbs
- Schematic of the apical ectodermal reach
- Signals come from the AER to the PZ and maintain the cells in the proliferative state
- First cells that differentiate and move out of the progress zone (blue cells) start to form the most proximal region of the
limb (stylopod), and so on to form the rest of the limb (zeugopod and autopod)
During outgrowth of the limb, cells of the PZ are assigned progressively more distal positional identities
Cells leaving the PZ differentiate and their positional value will depend on the time at which they leave the progress zone
The first cells leaving the PZ form proximal structures, the cells that have undergone numerous divisions in the PZ become the more
distal structures
As the bud grows, cells leave the PZ and start to differentiate, and cartilaginous structures begin to appear in the mesenchyme
The part of the limb nearest to the body wall is the first to differentiate, and differentiation proceeds distally as the limb extends
Phocomelia
Thalidomide (sedative) prescribed in the 60s causes severe limb deformities if taken at critical times during pregnancy
The long bones are shorter than normal and the more proximal elements are lost
Anterior-posterior axis of the limb
The anterior-posterior axis is specified by the posterior mesoderm of the limb bud known as the zone of polarizing activity (ZPA)
The pattern of digit formation is dependent upon the activity of the ZPA
- The ZPA secretes retinoic acid and SHH (sonic hedgehog)
Anterior transplantation of ZPA induces duplication of the digits
 - If you take ZPA cells from one limb bud and transplant them into the anterior region of another limb bud, you’ll get two
ZPAs and double the limbs number
- Polydactyly — formation of supernumerary digits
Cell death plays a major role in sculpting the limb
- It is essential for joint formation and for separation of the digits
- This process of programmed cell death (apoptosis) is genetically programmed
- Cell death is required to remove the webbing of the hands/feet so that we can get separation of the digits
Syndactyly — abnormal persistence of soft tissue between the digits (fusion of the digits)
Derivatives of the endoderm
Endoderm is the deepest of the three germ layers
Digestive tract (part of), respiratory tract (part of), tonsils, thyroid, parathyroid glands, thymus, liver, pancreas, gallbladder
General concepts regarding development of the gut and associated organs
Endoderm forms lining/epithelia and secretory elements of the digestive tube and glands
Mesoderm (mesenchyme) surrounds the tube and forms connective tissue and smooth muscles for peristalsis
Gut associated organs are generated through reciprocal inductive interaction
Endoderm  mesoderm
The same concept applies to the respiratory tract (epithelial lining from endoderm, connective tissue from mesoderm)
Formation of the gut tube
Lateral plates of the mesoderm come together ventrally to cause lateral flexion and fusion ventrally to give rise to the gut
- Lateral flexion gives rise to the gut itself!!!
At the same time, there is a longitudinal flexion where the embryo which is initially relatively flat starts to bend cranially and
caudally in the ventral region
Regionalization of the gut tube
Pharyngeal gut — extends from the buccopharyngeal membrane to the respiratory
diverticulum (pharyngeal pouches)
Foregut — extends as far as the liver bud (esophagus, stomach, proximal half of the
duodenum, liver and pancreas)
Midgut — extends to the proximal part of the transverse colon (distal half of
duodenum, jejunum, ileum, cecum, and part of colon)
- Primary intestinal loop
Hindgut — extends to the cloacal membrane (bladder, urethra, and prostate gland, the
rest of the colon and rectum)
Regional specification of the gut tube is initiated by a concentration gradient of
retinoic acid
- This gradient causes the expression of transcription factors in the gut endoderm that will determine the identity of the
different gut regions
- Different regions of the digestive tract are specified by a group of HOX genes, which provide positional info to different cell
tissues
- Genes are differentially activated in different regions of the gut, giving rise to different structures
Development of the liver and pancreas
Development of the liver
- Arises from the hepatic diverticulum during the 3 rd week
- As the hepatic diverticulum extends out from the foregut into the surrounding mesenchyme, the mesenchyme induces the
endoderm to proliferate, branch, and form the glandular epithelium of the liver
Molecular basis of liver induction
- The entire foregut endoderm has the potential to form liver tissue
- However, this potential is blocked by inhibitory factors produced by surrounding tissues — ectoderm, non-cardiac
mesoderm, and notochord
- In the prospective hepatic region, FGF2 and BMP secreted by the cardiac mesoderm relieve this inhibition allowing liver
tissue to form
Development of the pancreas
- Ventral and dorsal pancreatic outgrowths arise from the endoderm just caudal to the stomach during the 4 th week
o They eventually fuse with each other
- Insulin secretion begins around the 5th month
- In contrast to the liver, the notochord promotes pancreas development by repressing SHH expression in the endoderm
Physiologic herniation
Intestinal loops (midgut) enter the extraembryonic cavity in the umbilical cord during the 6 th week
- Looping into the umbilical cord gives the body cavity more room to develop certain organs
Retraction of the intestinal loops is initiated during the 10 th week and completed by 12th week
Development of the respiratory system
The respiratory diverticulum (lung bud) appears around the 4 th week
Outgrowth from the ventral wall of the foregut
The appearance and location of the lung bud are dependent upon an increase in retinoic acid produced by adjacent mesoderm
Epithelium of the internal lining of the larynx, trachea, and bronchi, as well as that of the lungs, is entirely of endodermal origin
The cartilaginous, muscular, and connective tissue components of the trachea and lungs are derived from splanchnic mesoderm
surrounding the foregut
Stages of pulmonary development
- Embryonic  pseudoglandular  canalicular  saccular  alveolar
One of the last systems to develop
Embryo stages

4th week
- Kind of see the heart
- See branchial arches
5th week
- See limb buds on forelimb and hind-limbs
6th week
- See umbilical cord  very large because some of the gut is inside the cord
th
8 week

Genera embryology IV
Development of the fetus
Fetal membranes and placenta
Birth defects
Prenatal diagnosis
Timeline of human development
Total fetal development takes 38 weeks from the time of fertilization
Embryonic period — up to week 8
Fetal period — after the first 8 weeks
Now talking about the fetal period
Development of the fetus
Fetal period is characterized by:
- Maturation of tissues and organs
- Rapid growth of the body (length and weight)
- Growth of the head slows
o Ratio of head to body decreases from 1:1 to 1:3 to 1:4
11-week human fetus
- 5-7 cm (crown-rump length)
- Swelling of the umbilical cord
- Digits are well developed
- Irregular skull contours
- Eyes ventrally located
12-week human fetus
- ~ 8 cm (crown-rump length)
- Primary ossification centers formed
- External genitalia recognizable
 - Vascularization visible
- Retraction of herniated loops
- Muscular activity (first signs)
18-week human fetus
- ~15 cm (crown-rump length)
- Eyebrows and head hair visible
- Outer ears in their final position
28-week human fetus
- ~27 cm (crown-rump length)
- Well-rounded contours
- Most organ systems are functional
- Except respiratory and nervous system
- Birth at 7 months = 90% survival chance
Feta membranes and placenta
The placenta is the primary site of nutrient and gas exchange
Functions — protection, nutrition, respiration, excretion, hormone production
Placenta is a fetomaternal organ with 2 components:
- Fetal component (chorion) derived from the trophoblast and extraembryonic mesoderm
- Materal component derived from the uterine endometrium
As the fetus grows, there’s an increase in nutrients demand
Facilitated by an increase in surface area between maternal and fetal components
Development of chorionic villi
Chorion is the embryonic derived portion of the placenta
- As the trophoblast grows it surrounds maternal blood vessels that are already present
- These blood vessels then fuse between spaces in the trophoblast to form a network of sinuses filled with maternal blood
- Even through the blood in the sinuses flows from the maternal blood, through the sinuses, and back into the mothers blood
supply it is important to note that the sinuses are not blood vessels and do not have a lining of endothelial cells to contain
the blood
- The blood comes in direct contact with the trophoblast cells which surround the growing embryo
- Eventually the trophoblast cells form fingerlike structure of the placenta composed of embryo-derived trophoblast cells
known as chorionic villi
Human blastocysts

Can see blastocysts at 7.5 days, 9 days, 12 days, and 13 days

By day 12, the trophoblast is expanding into the endometrium of the uterine wall
By day 13, the microvilli increase surface area with the mom; have primary villi
Primary villi = syncytiotrophoblast and cytotrophoblast
Seconday villi will have a mesoderm core in addition to the components of the primary villi
Tertiary villi has some developed vasculature in the mesoderm; occurs at day 21
Beginning of the 2nd month
- At first the chonionic villi are present around the entire blastocyst, but eventually as the embryo grows, they will only be
maintained around the region where the embryo is anchored and the others around will degenerate
Relation of fetal membranes to the wall of the uterus

Decidua basalis — component of the endometrium that will contribute directly to the placenta
Decidua parietalis — abembryonic region of the endometrium opposite the decidua basalis
Decidua capsularis — covers the embryo but will eventually degenerate
- Related to the chorion
Chorion frondosum — involved in formation of the placenta; opposite the abembryonic pole/chorion leave
Chorion laeve — fuses with decidua parietalis and the amnion (covers the embryonic cavity)
- Get complete loss of decidua capsularis here
Placenta forms from the fusion of villi that were maintained (e.g. chorion frondosum) and decidua basalis (endometrium)
Birth defects
Birth defect = congenital malformation = congenital anomaly
- Structural, behavioral, functional, and metabolic disorders present at birth
Birth defects are the leading cause of infant mortality
Major anomalies occur in ~3% of live-born infants
- Major survival implication
Minor anomalies occur in ~15% of newborns
Genetic factors account for approximately 28% of birth defects
Environmental factors produce approximately 3% to 4% of birth defects
Multifactorial inheritance produces 20% to 25% of birth defects
In 40% to 45% of birth defects the cause is unknown
Risk of birth defects being induced is greatest at 5th week of gestation, during the embryonic period (i.e. within the first 8 weeks,
before the first prenatal visit)
Types of anomalies
- Disruptions — morphological alterations of already formed structures
- Deformations — results from mechanical forces that mold a part of the fetus
- Syndromes — a group of anomalies occurring together that have a common cause
Environmental factors
- Infectious agents, radiation, chemical agents, nutritional deficiencies, maternal disease
- 1941 — discovery that German measles affecting a mother during early pregnancy caused abnormalities (cataracts and
heart defects) in the embryo
 - 1961 — first report linking limb defects to a sedative thalidomide, evidence that drugs could also cross the placenta and
produce birth defects
In 2010, the (CDC and the National Birth Defects Prevention Network published a study updating the national prevalence estimates
for selected birth defects in the United States from 2004-2006
- Orofacial defects (cleft palate/lip) are a major craniofacial defect
- Other major defects are cardiovascular defects (truncus), gastrointestinal defects, CNS defects (spinal cord closure), and
chromosomal abnormalities (trisomy 21)
Turners syndrome
- Mostly females
- Missing X chromosome
- Infertile
- Webbed neck
Prenatal diagnosis
Ultrasonography
- Noninvasive technique that uses high-frequency sound waves reflected from tissues to create images
- Transabdominal or transvaginal (higher image resolution)
- Performed between 18 and 20 weeks
- Assess overall growth and development, and birth defects
Maternal serum screening
- Concentrations of serum α-fetoprotein (AFP)
- AFP is produced normally by the fetal liver, peaks at approximately 14 weeks, and pass into the maternal circulation via the
placenta
- AFP concentrations increase in maternal serum during the second trimester and then begin a steady decline after 30 weeks
of gestation
- Test is performed between 15 and 20 weeks of gestation
- AFP can be used in combination with other markers such as unconjugated estriol (uE3), human chorionic gonadotropin
(hCG) and inhibin A
AFP uE3 hCG Inhibin A
Neural tube defects  - - -
Down syndrome    
Trisomy 18    -
Amniocentesis
- Approximately 20-30 ml of fluid is withdrawn from the amniotic cavity
- Procedure is performed transabdominally, in the second trimester (15-20 weeks)
- The fluid is analyzed for biochemical factors, such as AFP
- Fetal cells are recovered in the amniotic fluid  karyotyping (several weeks)
- Polymerase chain reaction can be performed to test for specific genetic diseases
Chorionic villus sampling
- Collection of approximately 5-30 mg of villus tissue
- Performed in a transabdominal or transcervical manner (between 10 and 12 weeks)
- Detection of genetic disorders and chromosomal anomalies (results in few days)
- This method does not assess neural tube defects or other malformations