Chapter CD

Some Miscellaneous
Topics
SOME RECENT ADVANCES IN EMBRYOLOGY
Recent advances in science and technology, especially in genetics and molecular biology, have
helped us to understand embryological development more clearly.
The uses of ultrasonography in the monitoring of normal and abnormal development;
and the use of techniques of in vitrofertilization (used in treatment of infertility) have helped
us tremendously.
Similarly, it is hoped that in near future we shall be able to use embryonic stem cells in the
treatment of various degenerative, genetic and malignant diseases.
The relationship between genetics and embryology is best reflected in gametogenesis.
The reduction in the number of chromosomes (haploid number) during gametogenesis and
its restoration (diploid number) at the time of fertilization has helped us to understand the
mode of inheritance of characters from parents to children. The anomalies in the number and
structure of chromosomes lead to various birth defects and syndromes. The phenomenon of
non-disjunction, which is due to abnormal mitosis, results in conditions like Turner syndrome,
and Downs syndrome.
Recent advances in molecular biology have been considered in detail in Chapters CD3
and CD4. They have helped us to understand the molecular control of development. Now we
know that growth and development is totally under the control of genes. Very recently we have
started understanding the role of specific genes in development (i.e., how, when and where
selected genes are expressed in an embryo to develop a particular structure). For the first time
we have understood the role of growth factors and other signaling molecules in embryonic
development. Similarly, we have started understanding the critical role of transcription
factors including HOX genes in the development of the embryo.
The knowledge of molecular biology is now helping us to understand the cause and
mechanism of congenital malformations. At present we know about many mutant genes that
are responsible for various kinds of birth defects.

Phases of the Cell Cycle

One cell cycle consists of the events taking place between the end of one cell division up to the
end of the next cell division. The cell cycle may be divided into four stages. These are the G1,

Human Embryology

Fig. CD-5.1: The cell cycle consists of four phases i.e., G1 (Gap phase or pre-synthetic phase); S phase
(synthetic phase in which DNA replicates); G2 phase (chromosome begins to get condensed and two sister
chromatids are formed) and the M phase in which the cell divides into two daughter cells. Sister chromatids
split at the centromere and move into the two daughter cells.

S1, G2, and M phases. The G1, S1 and G2 phases are collectively known as interphase. This
is the phase between two successive mitoses during which a cell duplicates its DNA contents
and prepares itself for next cell division. In the M phase (mitosis phase) a cell divides into two
daughter cells. Figure CD-5.1 shows the synthesis of DNA during interphase and the splitting

Fig. CD-5.2: Summary of events occurring during one cell cycle.

Chapter CD 5 Some Miscellaneous Topics

of sister chromatids during the M phase of the cell cycle. Fig. CD-5.2 presents the summary of
events occurring during the interphase and the M phase of a cell cycle.

Inter-dependence of Oocyte and Follicular Cells

Both the oocyte and surrounding follicular cells are dependent on each other for further
development. The follicular cells secrete meiotic inhibitory factors, which prevent primary
oocytes from maturing beyond the prophase of the first meiotic division. This effect can last
for many years. These inhibitory factors are transmitted to the oocyte via gap junctions present
between microvilli of oocytes and of follicular cells. These junctions lie in the zona pellucida
(Fig. CD-5.3). Follicular cells are also responsible for growth, metabolism and maturation of
oocytes. Conversely, oocytes are responsible for proliferation and differentiation of follicular
cells. Factors produced in the oocyte help in formation and maturation of Graafian follicles.

Prechordal Plate
The prechordal plate is a small collection of cells near the cranial end of the notochord. The
cells in it are apposed to endoderm. These cells influence the formation of forebrain and eye.

The prechordal must not be confused with the prochordal plate.

Cranial migration of cells from the primitive knot and into the notochord occurs in two
phases.
1. The first phase of migration forms the prechordal plate.
2. The second phase of migration of cells forms the notochordal process.

Fig. CD-5.3: Diagram to show the intimate relationship of oocyte and follicular cells (Gap junctions are
present microvilli of the two.

Human Embryology

SOME CHROMOSOMAL ABNORMALITIES

Trisomies associated with autosomes:

Trisomy-21,
Trisomy-13,
Trisomy-18,

Downs syndrome
Pataus syndrome
Edwards syndrome

Disorders of Chromosome Structure

The most common abnormalities refer to chromosomes in which a portion of the chromosome
is missing or is represented twice. The number of chromosomes is normal (46) but the structure
of one or more chromosomes is abnormal. The abnormalities in structure of the chromosomes
results because of chromosome breakage. Some important structural abnormalities are as
follows.
Deletion
Inversion
Ring chromosome
Isochromosome
Translocation
Duplication
Ring shaped chromosome.

These abnormalities are responsible for various disorders. This occurs because of missing
genes (as in deletion), gain of genes (as in duplication), and also because of change in normal
position of genes (as in inversion and translocation). Well-known structural abnormalities are
associated with the deletion and translocation.

The following is a list of common syndromes associated with deletion:

Syndrome
Cri-du-chat
Prader-willi
Angelman
Wilms tumour
Miller Diekar
Di-George

Affected chromosome number

5 (5p-)
15
15
11
17
12

All the above syndromes are associated with many birth defects and mental retardation.
Almost about 4% cases of Downs syndrome are due to Robertsonian translocation.

How chromosomal abnormalities (numerical and structural) are detected in

laboratories
Chromosomal analysis is required in many clinical conditions for proper diagnosis of genetic
diseases. The chromosomal constituent of an individual is called the karyotype. Karyotyping

Chapter CD 5 Some Miscellaneous Topics

is a procedure to obtain the karyotype of an individual. With the help of a karyotype, numerical
and structural abnormalities of chromosomes can be easily identified. The analysis of
chromosomes becomes very precise with the help of various banding techniques (G-banding,
Q-banding, R-banding etc.).
Fluorescent in situ hybridization (FISH) is a new diagnostic technique that uses a
specific DNA probe to identify a specific chromosome. With the help of multicolor spectral
karyotype (SKY), all chromosomes appear in colour and a multi-color karyotype is obtained.
This technique is used to detect chromosomal abnormalities like deletion or translocation.

Gene Abnormalities (Gene Mutations)

Gene mutations are also responsible for various kinds of hemoglobinopathies (thalassaemia
and sickle cell anemia etc) and inborn errors of metabolism (alkeptonuria, phenylketonuria,
albinism, galactosemia etc.).

Fertilization
1. A very large number of capacitated spermatozoa succeed in passing through the cells of
the corona radiata and reaching the ovum. However, only one of them is able to fertilize
the ovum.

The glycoprotein of the zona pellucida is responsible for induction of the acrosomal
reaction. The release of acrosomal enzymes (acrosin) helps the spermatozoon to penetrate
the zona pellucida.

The zona pellucida contains three different glycoproteins ZP1, ZP2 and ZP3. The sperm
head contains a specific binding site for ZP3. The sperm-binding site of the ZP3 molecule
varies from species to species. This explains why the spermatozoa of one species cannot
fertilize the egg of other species.

In Vitro Fertilization
The technique of in vitro fertilization includes the following steps:
Stimulation of production of oocytes by administration of the drug clomiphene or
gonadotropin.
Oocytes are collected by a laproscopic technique.
Spermatozoa are are capacitated by treating them with certain ionic solutions.
Oocytes and spermatozoa are kept in a petri dish containing a special culture medium.
The fertilization and subsequent cleavage is monitored microscopically.
Alternatively, an oocyte may be fertilized by microinjecting it with a spermatozoon.
One or more embryos, (that have reached the eight cell stage, or have become a blastocyst),
are transferred to the uterus using a catheter.
The remaining embryos, which are obtained from in vitro fertilization, but are not used,
can be stored for future use by freezing (cryo-preservation) for long periods.
Only about 20 to 30% of embryo transfer attempts result in successful pregnancy.

Human Embryology

SOME CONGENITAL ABNORMALITIES

Caudal Regression Syndrome (Sirenomelia)
This condition is characterized by the absence of the sacrum and dysgenesis of the lower
lumbar vertebrae. There may be dysgenesis and fusion of the legs. Other anomalies associated
with the condition are renal agenesis, imperforate anus and abnormalities of genital organs.

Sirenomelia results due to disruption of gastrulation. As a result insufficient mesoderm
is formed in the caudal most region of the embryo. As a result caudal parts of the body (lower
limb, lower vertebrae and urogenital system) are malformed. Abnormal gastrulation occurs
either because of genetic abnormalities, or due to the effects of teratogens.

Some Known Disorders of Craniofacial Development

Many syndromes are associated with disorders of craniofacial development. Some of these
are:
Robin sequence,
Mandibulo-facial dysostosis (Treacher Collins syndrome),
Apert syndrome, and
Crouzon syndrome.
These are genetic disorders, which are also termed the branchial arch syndromes. It is
believed that neural crest cells are essential in the formation of the craniofacial region. Failure
of the development of neural crest cells may lead to defective formation of the craniofacial
region. The first branchial (or pharyngeal) arches (which give rise to the maxilla and the
mandible), are particularly affected. This results in mandibulo-facial dysostosis also called
the Treacher Collins-Franceschchetti syndrome.

Sternum
When the fusion of the two sternal bars is faulty, the body of the sternum shows a partial or
even a complete midline cleft. This is due to mutation in the Hoxb-2 and Hoxb-4 genes. Minor
degrees of non-fusion may result in a bifid xiphoid process or in midline foramina. Transverse
clefts may also occur. Malformation of the xiphoid process is due to mutation in Hoxc-4 and
Hoxa-5 genes.

Achondroplasia
Achondroplasia is an autosomal dominant trait. It is a skeletal growth syndrome characterized
by short stature due to slow development of the middle portions of the long bones in the
arms and legs. The most common form of achondroplasia is due to a defect of the Fibroblast
Growth Factor Receptor (FGFR), and is recognised by exaggerated cranial growth and bossing
and depression at the bridge of nose. A second form is pseudoachondroplasia, which is due
to defect of Cartilage oligomeric Matrix Protein (COMP) in the joints and is characterized by
more typical development of head and face.

Chapter CD 5 Some Miscellaneous Topics

Cleidocranial Dysostosis
The formation of the clavicle is of particular importance as the bone ossifies in membrane
and its developmental anomaly results in a syndrome called Cleidocranial dysplasia. The
condition is due to mutation in Cbfa1 gene (core binding factor alpha-1) which causes
hypoplasia and delayed ossification of clavicle and other membrane bones of skull. The person
may also show the presence of supernumerary teeth. The gene CBFA1 is located on the short
arm of chromosome number 6. CBFA1 gene controls differentiation of precursor cells into
osteoblasts and is thus essential for membranous as well as endochondral bone formation.

SOME FACTS ABOUT THE NEURAL CREST

1. It is believed that neural crest cells are formed due to action of bone morphogenetic
proteins (BMP-4/7) and Wnt, which are secreted by neighboring ectoderm.
2. Neural crest cells are divided mainly into three divisions viz. cranial neural crest,
circumpharyngeal neural crest and trunk neural crest. The neural crest cells of the
above three divisions follow different migratory pathways (Fig. CD-5.4).
a.
The cranial neural crest arises in the forbrain, midbrain and anterior rhombencephalic
regions. Cells arising from the cranial part of the neural crest are believed to
migrate into the mesenchyme of the head and neck and influence development of
somitomeres. They play an important role in the development of bones, connective
tissue and musculature of the head, and face.

Fig. CD-5.4: Various neural crest derivatives and migratory paths in the trunk. Neural crest cells migrate
through a dorso-lateral pathway to form pigment cells. They migrate through a ventro-lateral pathway to
form dorsal nerve root ganglia. They also migrate through a ventral pathway to form sympathetic ganglia,
preaortic ganglia and parasympathetic ganglia in the gut wall. They also form the adrenal medulla.

Human Embryology

b.
The circumpharyngeal neural crest arises in the posterior rhombencephalic region
and in the lower part of the pharynx. The neural crest cells from the anterior
rhombencephalon migrate towards the developing heart and aortic arches where
they contribute to the formation of the outflow tract of the heart, and of great vessels.
The neural crest cells from the levels of somites 1 to 7 migrate towards the developing
gut and form parasympathetic neurons and myenteric plexuses, which innervate the
digestive tract.
c.
The neural crest cells of the trunk arise from the level of the sixth somite to the level
of the last somite. These cells migrate dorsolaterally to skin, and form melanocytes.
Some cells also migrate ventro-laterally to form sensory ganglia. Ventrally they
migrate to form sympathetic ganglia, the adrenal medulla, the pre-aortic ganglion and
the parasympathetic ganglia and plexuses in the gut wall.

Neural Crest and Spiral Septum

The neural crest cells contribute to the formation of the spiral septum (in the truncus arteriosus
and the outflow tracts of the right and left ventricles). These neural crest cells migrate from
the region of the hindbrain to the developing heart. They pass through pharyngeal arches 3,
4 and 6. Failure of such migration of neural crest cells leads to congenital malformations of
the outflow tract of the heart i.e., pulmonary stenosis, teratology of Fallot, persistent truncus
arteriosus and transposition of great vessels. As the neural crest cells also contribute to the
formation of various parts of the head and neck congenital anomalies of the heart and of the
head and neck are often associated with each other.