2B Dev Bio

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SPERMATOGENESIS
Spermatogenesis is the multi step process of the male gamete formation (sperms) from primordial
germ cells in the male gonads of animals.
Spermatozoon: Functional sperm cell, especially in animals where the sperm cell is flagellated and
hence, motile.
Developmental Process
Once the vertebrate primordial germ cells arrive at the genital ridge of a male embryo, they become
incorporated into the sex cords and remain there till maturity
At maturity, sex cords become internally hollow and form seminiferous tubules. Spermatozoa are
produced in semi-neferous tubules in the testes.
Epithelium of semi nefarious tubules differentiates into Sertoli cells, which nourish and protect
the developing sperm cells.
Spermatogonia line the seminiferous tubule, near the basement membrane
Spermatogonia originate at puberty by proliferation of Gonocytes and are descendants of
primordial germ cells.
Spermatocytogenesis: Creation of spermatocytes by proliferation and commitment.
Mitosis of Spermatogonia, which generate several lines of Spermatogonia and finally result in
primary spermatocytes
Primary Spermatocytes undergo meiotic division to yield secondary spermatocytes
Spermidatogenesis: Secondary spermatocytes complete the second division of meiosis and produce
haploid cells, called Spermatids.
Spermiogenesis: Creation of spermatozoa through further development and metamorphosis of
spermatids. Results in functional sperms.
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Spermatocytogenesis
Starts with division of spermatogonia that line the

seminiferous tubules, near the basement membrane
PGCs divide into A1 Spermatogonia (smaller than PGCs,
ovoid nucleus)
A1 has self renewal capacity like stem cells
A1 — A2 — A3 — A4 (Possible that all of them can self
renew)
A4 has three possible fates
Self renew into A4
Cell Death by Apoptosis
Intermediate Spermatogonium
Intermediate Spermatogonium
First committed stem cell type
Committed to become spermatozoa
Type B Spermatogonium
Precursors of Spermatocytes
Last cells in the line that undergo mitosis
Primary Spermatocytes
Cells that enter meiosis
Secondary Spermatocytes
Primary Spermatocytes undergo the first meiotic
division to yield secondary spermatocytes
Features
During spermatogonial divisions, cytokinesis is not

complete.
The cells form a syncytium where each cell communicates
with others via cytoplasmic bridges.
Because ions and molecules readily pass through these
intercellular bridges, each cohort matures synchronously.
During Spermatocytogenesis, spermatocyte nucleus often
transcribes genes whose products will be used later to form
the axoneme and acrosome.
Spermatidogenesis
Secondary spermatocytes undergo Meiosis II and give rise

to oval, non-flagellated haploid cells, called Spermatids
During the division from A1 spermatogonium to
Spermatid, the cells move farther and farther away from the
basement membrane of the seminiferous tubule, and closer
to its lumen. — Each type of cell can be found in a
particular layer of the tubule.
Spermatids are located at the border of the lumen. Here
they lose their cytoplasmic connection and differentiate
into sperm cells.
Spermiogenesis

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Stage of differentiation of the sperm cell, during which it is

prepared for functions of motility and interaction towards
the egg.
Construction of acrosomal vesicle from the Golgi
apparatus. The acrosome forms a cap that covers the sperm
nucleus.
As the acrosomal cap is formed, the nucleus rotates so that
the cap will be facing the basal membrane of the
seminiferous tubule. The rotation is necessary to allow the
flagellum to develop towards the lumen
Flagellum develops from centriole on the other side of the
nucleus, into the lumen
In the late stages of spermiogenesis, the nucleus flattens
and condenses.
Histones are replaced by protamines.
Transcription of gene for protamine is seen in early
haploid cells (spermatids), but translation is delayed.
During spermiogenesis, the nucleosomes dissociate
and histones are replaced by protamines.
This causes shutdown of transcription in the nucleus,
and facilitates an almost crystalline structure.
After nucleus condensation, most of the sperm cell
cytoplasm is discarded as cytoplasmic droplet.
Mitochondria form a ring around the base of the flagellum.
This is where most of the ATP for Sperm cell motility is
generated.
Resulting sperms enter the lumen of the seminiferous
tubule.
Control of Spermatogenesis
Control of Initiation
Initiation of spermatogenesis during puberty is regulated by synthesis of BMP8B by the
Spermatogonia.
When BMP8B reaches a critical concentration, the germ cells begin to differentiate.
Differentiating cells produce high levels of BMP8B, which can further stimulate their
differentiation.
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Mice lacking BMP8 do not initiate spermatogenesis at puberty

Control of Commitment
Glial Cell Derived Neurotrophic Factor (GDNF) causes Spermatogonia to take the path towards
differentiation rather than self renewal.
GDNF is secreted by Sertoli cells. Levels decide whether a dividing Spermatogonia remain
spermatogonia (low levels) or enter the pathway to become spermatocytes (high levels).
GDNF is regulated by FSH — GDNF provides a link between Sertoli cells and Endocrine
system. It is a mechanism for FSH to instruct testes to produce more sperm.
Overall Hormonal Control
Hormones produced by Testes that directly or indirectly affect spermatogenesis — testosterone,
estradiol and inhibin
Testosterone by Leydig cells located adjacent to seminiferous tubules
Estradiol and Inhibin by Sertoli cells within Seminiferous tubules (which also produce
GDNF that controls commitment of Spermatogonia)
Development of Spermatogenesis at Puberty, depends on the Hypothalamus, Pituitary Gland and
functional Leydig cells (to produce Testosterone)
In the absence of Pituitary gland, spermatogenesis can be initiated by FSH and Testosterone
FSH is necessary to develop androgen binding protein production by Sertoli cells
Once the Sertoli function is developed, Testosterone alone can maintain spermatogenesis.
Yield of Spermatozoa is increased if FSH is present.
This is because FSH prevents atresia of differentiating A type spermatogonia
Loss of spermatogonia can also be reduced by increased sexual activity.
FSH levels are decreased by Inhibin
Composition of Semen
Semen is a translucent white coloured fluid with a density 10x water. In normal male, volume of
ejaculated semen is 2 to 6 ml.
pH 7.2 to 8.0
Sperm concentration of 20 x 10^6 spermatozoa/ml.
Spermatozoa (10%) + Variety of compounds and enzymes in the seminal plasma (90%) which help in
nourishment, survival and normal functioning of the spermatozoa.
Organ % Components
Approximately 200 to 500 million Spermatozoans are released per

Testes 2-5
ejaculation.
Amino acids, Citrates, Enzymes, Flavins, Proteins, Vitamin C

Fructose (main energy source of sperm cell, which rely entirely on
sugars for energy)
Seminal Vesicle 65-75 Prostaglandins (suppress immune response by female against the
semen)
Fluid from seminal vesicles and mucous glands gives the semen a
mucoid consistency
Prostate 25-30
Acid Phosphatase, Citric Acid, Prostate specific antigen, Proteolytic
enzymes
Zinc (stabilises the DNA in sperm cells. Zinc deficiency may result in
lowered fertility because of increased sperm fragility. It can also
adversely affect spermatogenesis.)
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Prostatic fluid gives semen a milky appearance.

Clotting enzyme from Prostatic fluid causes the Fibrinogen of the
seminal vesicle fluid to form a weak fibrin coagulum that holds semen
in deeper regions of the vagina where the uterine cervix lies.
Coagulum dissolves in the next 15 to 30 minutes due to lysis by
fibrinolysis. As the coagulum dissolves, the sperm becomes highly
motile.
Galactose,
Bulbourethral Mucus (creates a less viscous channel for sperms to swim through in
<1
Glands vagina and cervix, and prevent their diffusion out of semen)
Sialic Acid
Sperm Capacitation
Capacitation is a process of functional maturation of the Spermatozoon through cellular and molecular
changes that allow the Spermatozoon to go through Acrosome reaction. It is largely a biochemical
maturation process of the sperm cell membranes and motility properties.
Though Spermatozoa are mature when they leave the epididymis, their activity is held in check by
multiple inhibitory factors secreted by male genital duct epithelia.
When they are first expelled in the semen, they are unable to perform their duties in fertilising the
ovum.
On coming in contact with the fluids in the female genital tract, changes occur that activate the sperm
for the final processes of fertilisation. (Collectively called Capacitation). Requires 1 to 10 h in humans.
Non-mammalian spermatozoa do not require the capacitation step and are ready to fertilise an oocyte
immediately after release
Process of Capacitation
When spermatozoa remain in the fluid of the male genital ducts

Continually exposed to many floating vesicles from the seminiferous tubules containing large
amounts of cholesterol.
This is continually added to the membrane covering the acrosome, toughening the membrane and
preventing release of its enzymes
Sperm head is covered with some glycoproteins which do not allow sperm cell membrane
receptors to be exposed
Inside the female tract, Destabilisation of sperm head membrane, rendering it more fusigenic
removal of sterols and and non-covalently bound seminal glycoproteins, which is made possible
by secretions of sterol binding albumin, lipoproteins, proteolytic and glycosidasic enzymes like
Heparin by the Uterus.
Sterol removal leads to a more fluid membrane
Area of the acrosomal cap is altered so that acrosome reaction becomes possible.
Sperm cell membrane receptors and transporters are exposed due to removal of Glycoprotein
layer.
Due to transporter activation, membrane has an increased permeability to Ca2+. Influx of Ca
increases chances of membrane fusion.
Calcium ions also cause changes in membrane of acrosomal cap, making ti possible for the
acrosome to release its enzymes rapidly as the sperm penetrates the granulosa cell mass and zona
pellucida surrounding Ovum.
Swimming Properties
Essentially outcome of modifications in membrane of the sperm cell.
Hyperactivity: whipping movements of tail with larger sideways swimming movements of the
head. It is related to influx of Calcium ions which produce increased intracellular cAMP
concentrations, and thus increase motility.
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Calcium also directly provides a powerful whiplash motion to the flagellum instead of the weak
undulating motion
Uterine and Fallopian tube fluids wash away inhibitory factors that suppress sperm activity.
Elevated cAMP activates Fertilisation Promoting Factor produced by the male.
Synchrony of Capacitation
Cannot be determined ahead when exactly the Oocyte and Spermatozoon will meet
Maturation mechanisms are so configured that various groups of sperm cells are able to keep their
chances of fertilisation upright over a relatively long time after cohabitation.
Ejaculated sperm cells do not all undergo capacitation at the same time — heterogenous groups.
Ensures that atleast some groups of sperm cells are capable of fertilisation at all times.
In-Vitro Capacitation for IVF
Semen collected is filtered through gauge to remove collecting gel (if used to collect semen) and debris
2ml of Semen is transferred to a pre-warmed 15 ml tube, mixed with 6ml Tyrode medium, centrifuged
at 900xg for 10 minutes to allow removal of seminal plasma.
Supernatant is removed and pellet is resuspended in modified Tyrode medium + bicarbonate medium to
a final volume of 2ml. Incubated at 37ºC for 30 min.
Test of capacitation is done by Chlortetracycline staining and Merocyanine staining. In Merocyanin,
stains are darker if the lipids are in higher state of disorder as is in the case of Capacitated
Spermatozoa.
After capacitation, sperm and egg are incubated together (75000:1 ratio) in culture media for about 18
hours.
OOGENESIS
Oogenesis is the process formation of a functional female gamete from immature ova. In mammals, the
process occurs in the ovarian follicle of the ovary (Graafian follicles)
Graafian follicle contains
External fibre-vascular coat

Internal coat of nucleated cells
Transparent, albuminous fluid in which the ovum is suspended.
Steps in Oogenesis
Sexual differentiation of Primordial Germ Cells: Mostly occurs in the embryonic stage. Resulting cells
are Oogonia
Oocytogenesis: Development of primary oocytes from oogonial cells. Mostly in embryonic stage
Primary Maturation Division: Gametogenic Meiotic division I, results in a secondary oocyte and a
polar body
Secondary Maturation Division: Gametogenic Meiotic division II, results in an Ootid.
Ootid maturation: Process of cellular differentiation, results in an Ovum.
Why is Oogenesis different from Spermatogenesis?
Gamete formed by spermatogenesis is essentially a motile nucleus, while the gamete formed by
oogenesis contains all the material needed to initiate and maintain metabolism and development.
In addition to forming a haploid nucleus, oogenesis also builds up a store of cytoplasmic enzymes,
mRNA, organelles, metabolic substrates

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Mechanisms of oogenesis vary among species more than spermatogenesis, since patterns of
reproduction vary so greatly between species.
Oogenesis in Amphibians
In frogs and other species that produce thousands of eggs, the oogonia are self-renewing stem cells that
endure for the lifetime of the organism
Oogenesis takes 3 years roughly.
2 years: oocyte increases its size very gradually
3rd year: Rapid accumulation of yolk in the oocyte causes the egg to swell to a large size
Eggs mature in yearly batches. The first cohort matures shortly after metamorphosis
Two principal events go on side by side
Meiotic cell division: Formation of haploid cells from an original diploid cell (i.e. primary oocyte
contained in female ovaries)
Materials get stored: Energy sources and energy generating organelles (yolk and mitochondria),
enzymes and precursors for DNA, RNA and protein synthase, stored mRNA, structural proteins,
morphogenetic regulatory factors.
Meiotic Divisions
Meiotic division starts in the first year of the Oogenesis. But once the process reaches the Diplotene of
Meiotic Prophase I, it slows down greatly.
Amphibian oocytes can remain for years in the Diplotene stage of Meiotic prophase. Resumption of
meiosis in the amphibian primary oocyte requires Progesterone (secreted by follicle cells in response to
gonadotrophin hormones secreted by the Pituitary gland)
Progesterone — MPF protein (M Phase promoting factor) which contains cyclin and cyclin dependent
kinases which form regulatory complexes for cell division
Within 6h of Progesterone stimulation, NE disintegrates — Germinal Vesicle Breakdown
Nucleoli disintegrate and chromosomes contract and migrate to the animal pole to begin division.
First meiotic division is completed and mature ovum is released from the ovary — Ovulation
Oogenic Meiosis
Unique in its placement of the metaphase plate.

When the primary oocyte divides, the nucleus breaks down and the metaphase spindle migrates to the
periphery of the cell.
At telophase, one of the two daughter cells contains hardly any cytoplasm, whereas the other has nearly
all the cytoplasmic volume.
Smaller cell is the First Polar Body and the large cell is called the Secondary Oocyte
During 2nd division of Meiosis, a similar unequal cytokinesis takes place — most of the cytoplasm is
retained by the mature egg (ovum) and a second polar body receives very little cytoplasm and a haploid
nucleus.
Thus, oogenetic meiosis conserves almost the entire volume of oocyte cytoplasm in a single cell rather
than splitting it equally among 4 progeny
Yolk Development
Occurs in the Diplotene stage of Meiotic prophase

Yolk is a mixture of materials used for embryonic nutrition.
The major yolk component is a 470 kDa protein called Vitellogenin
Synthesised in the liver and carried by the bloodstream to the ovary.
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Passes between the follicle cells of the ovary, and is incorporated into the oocyte by
micropinocytosis
In mature oocyte, vitellogenin is split into — heavily phosphorylated Phosphovitin and
lipoprotein Lipovitellin. They are packaged together into membrane bound yolk platelets.
Yolk deposition occurs in a differential manner, due to which eggs are highly asymmetrical (animal-
vegetal axis). Concentration of yolk is very high towards the vegetal pole and very low in the animal
pole.
As the yolk is being deposited, organelles also become arranged asymmetrically. Cortical granules
form the GA, which are initially scattered throughout the cytoplasm but later migrate to the periphery
of the cell.
Mitochondria divide rapidly, forming millions of mitochondria that will be distributed among the
different blastomeres during cleavage.
Cortical granules, mitochondria, pigment granules are found at the periphery of the cell (in an actin rich
oocyte cortex).
In the inner cytoplasm, distinct gradients emerge
Yolk granules are heavily concentrated at the vegetal pole
Glycogen granules, ribosomes, lipid vesicles and ER are found towards the animal pole.
Specific mRNAs combined with proteins (informosomes) stored in the cytoplasm become
localised to certain regions of the oocyte. In frogs, mRNA are mostly synthesised by Lampbrush
Chromosome mechanism
Cytoskeleton plays a significant role in localising specific mRNAs and morphogenetic factors.
Oogenesis in Mammals
Two patterns of ovulation
Ovulation is stimulated by act of copulation

Physical stimulation of cervix — gonadotrophin from pituitary — signals egg to resume meiosis
— expel egg from ovary
Ensures that most copulations will result in fertilised ova
e.g. Rabbits
Estrus
Periodic ovulation pattern, in which the female ovulates only at specific times of a year
Menstrual cycle: Characteristic primate periodicity in maturing and releasing ova
It entails the periodic shedding of blood and endothelial tissue from the uterus at monthly
intervals.
Human females have cyclic ovulation, averaging once every 30 days.
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Menstrual cycle in humans is an integration of
Ovarian cycle — mature and release an oocyte

Uterine cycle — provide appropriate environment to the developing blastocyst
Cervical cycle — allow sperm to enter female reproductive tract only at appropriate time.
Oogenesis and Ovulation in humans
Early in embryogenesis, PGCs migrate from yolk sac endoderm to the genital ridge (developing ovary)
where they take up residence and are called oogonia (Diploid cells)
In the human embryo, about a thousand oogonia divide rapidly from 2nd to 7th month of gestation.
After the 7th month of embryonic development, most oogonia die.
Remaining oogonia enter the first meiotic division (primary oocytes) and progress until Diplotene
phase in which they are maintained till puberty. They grow in size during this arrested phase but do not
divide.
When female reaches sexual maturity, small number of primary oocytes are stimulated to continue
through Meiosis I, under influence of FSH
Each oocyte in adult human ovary I enveloped by a primordial follicle consisting of
Single layer of epithelial granulosa cells
Less organised layer of mesenchymal theca cells
When an oocyte is activated, it undergoes a great increase in volume. There is also an increase in
number of follicular granulosa cells, which form concentric circles around the oocyte. Proliferation of
granulosa cells is mediated by a paracrine factor GDF9 (member of TGFß family)
During this growth period, the oocyte is in resting stage. Fully grown follicle contains a large oocyte
surrounded by several layers of granulosa cells. Innermost layer stays with the ovulated egg forming
the cumulus, which surrounds the egg in the oviduct.
During growth of the follicle an Antrum forms, which becomes filled with a complex mixture of
proteins, hormones and other molecules.
Meiotic Division I is completed hours before ovulation (release of ovum from the ovary). Completion
occurs in mature Graafian follicles.
Diploid to Haploid
Asymmetric cytokinesis (smaller polar body contains half the chromosomes but a very small
amount of cytoplasm and eventually degenerates)
When primary oocyte completes the first meiotic division, it is called secondary oocyte (1N).
Ovulation occurs at this stage
If the secondary oocyte is not penetrated by a sperm, it will degenerate. If fertilisation and pregnancy
do not occur, new menstrual cycle will begin in which FSH from Pituitary will stimulate a few more
primary oocytes to continue through Meiosis I.
If a sperm penetrates the zona pellucida, secondary oocyte is stimulated to continue through Meiosis II,
forming secondary polar body and a mature ovum.
Fertilisation is complete when the two fuse and restore the chromosome number.

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Hormonal Control
Early hormonal control helps follicle to develop and forces oogenesis to occur in a cycle in a certain
time period
Control begins in Hypothalamus
Hypothalamus — GnRH — Anterior Pituitary — FSH, LH
Granulosa cells express FSH receptors — growth of follicle
Theca cells express L receptors — growth of corpus luteum
Theca cells also produce androgens, which granulosa cells convert to estrogen.
Estrogen again acts on Anterior Pituitary to further FSH and LH surges, and supports growth of
endometrium
Dominant follicle secretes Inhibin which acts on Pituitary to stop FSH. Only the dominant follicle, now
FSH independent will continue to grow
Granulosa cells increase FSH and LH receptors and Theca cells increase LH receptors — surge in
hormone reception results in ovulation
If fertilisation occurs — Corpus luteum secretes progesterone which supports growth of Endometrium
If fertilisation does not occur — Hormone levels drop, Corpus luteum breaks leading to drop in
Progesterone, Endoemtrium sloughs off — Menstruation

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FERTILISATION
Fertilisation is the process by which compatible gametes — male sperm and female egg — fuse
together, forming a single zygotic cell that develops into the next generation of the organism through
steps of embryogenesis
It begins when the sperm contacts the outer surface of the egg and ends when sperm nucleus fuses with
the egg nucleus
May vary from 30 minutes in sea urchins to several hours in mammals
After fertilisation, the fertilised egg is called zygote. Once zygote divides, it is called embryo
External Fertilisation
Egg and sperm come together outside of the parent’s bodies.

e.g. Sea Urchins, Starfish, Clams, Mussels, Frogs, Corals
Gametes are released (spawned) by the adults into a water body. Fertilisation takes place in this watery
environment, where embryos start to develop.
Disadvantage: Meeting of egg and sperm is left to chance.
Swift water currents, water temperature changes, predators, pollutants can prevent fertilisation
One of the important adaptations to counter these adversities is to produce millions of sperms and eggs,
so that fertilisation of even a small fraction would also result in a significant number of offspring
Males and females use behavioural clues, chemical signals and other stimuli to coordinate spawning so
that sperm and eggs are in spatial and temporal proximity
No parental care. Instead, eggs contain a food supply in the form of yolk that nourishes the embryo
until they hatch and are able to feed on their own
Internal Fertilisation
Takes place inside the female body

Male typically has a penis which delivers sperm into the female’s reproductive tract.
All mammals, reptiles and birds, and some invertebrates like snails, worms and insects use internal
fertilisation.
It does not necessarily require that the growing embryo remains inside the female body. e.g. In honey
bees, the queen bee deposits the fertilised eggs into special compartments in the honeycomb. These
compartment are provided with food resources for the bees to use as they develop
Because the sperm and egg are always protected inside the bodies of the male and female, and are
deliberately placed in close contact during mating, relatively few sperm and eggs are produced.
Extensive parental care to the animals
In mammals, two additional organs develop to support the growing embryo — uterus and placenta
Sequence of interaction of sperm and egg
Chemoattraction of the sperm to the egg by soluble molecules secreted by the egg.
Binding of sperm to the extracellular envelope (vitelline layer/zona pellucida) of the egg
Exocytosis of the acrosomal vesicle to release its enzymes
Passing of sperm through this extracellular envelope
Fusion of egg and sperm plasma membranes

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Fertilisation in Mammals
Sperm are attracted to the fluid surrounding the egg.

Chemotaxis
Exact nature of this fluid is not known
Zona pellucida in mammals plays a role analogous to vitelline envelope in

invertebrates
Zona pellucida is a glycoprotein matrix which is synthesises and secreted by
the growing oocyte.
Binds the sperm (Zona Protein 3 is the sperm binding glycoprotein of
Gamete Binding
the mouse zona pellucida. Set of proteins on the sperm can recognise
and Recognition
specific carbohydrate regions of ZP3)
Initiates Acrosomal reaction after the sperm is bound
Binding of sperm to the zona is relatively species specific. Species specific
gamete recognition is not a major problem when fertilisation occurs
internally
Acrosomal reaction in mammals occurs only after the sperm has bound to
zona pellucida
Mouse sperm acrosomal reaction is induced by cross linking of ZP3 with
receptors for it on the sperm membrane.
Cross linking opens calcium channels to increase the concentration of calcium
Exocytosis
in the sperm.
Increased concentration of calcium ions — fusion of acrosomal vesicle with
plasma membrane — exocytosis of acrosomal vesicle — release of
proteolytic enzymes — digests a path through zona pellucida to the egg
surface
During acrosomal reaction, anterior portion of the sperm PM is shed (where

ZP3 proteins are located)
Sperm must still remain bound to zona to lyse a path through it
Passing of sperm Secondary binding to zona is through proteins in inner acrosomal membrane,
that bind to ZP2
Structure of zona has repeating units of ZP3 and ZP2, occasionally cross
linked by ZP1
Fusion of PM
Lysis is followed by fusion of sperm PM with egg PM
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Fertilin proteins in sperm plasma membrane are essential for membrane

fusion.
In mice, fertilin adheres sperm to the egg by binding integrin on egg PM
After this, entire sperm cell content and components enter the egg including
mid piece and tail
Egg and sperm NE fuse, permitting chromosomes from the egg and sperm to
mix
Zygote is formed and development of embryo begins.
Prevention of Polyspermy
Fast Block to Polyspermy Slow Block to Polyspermy
Achieved by changing the electric potential Cortical granule reaction: slower, mechanical
of the egg PM block to polyspermy that becomes active about
Egg PM has resting membrane potential at a minute after the first successful sperm-egg
-70 mV (interior of cell is negatively attachment.
charged with respect to the exterior) Upon sperm entry, cortical granules beneath
In 1-3 s of binding of sperm, membrane the PM fuse with PM and release their
potential shifts to a positive level (20 mV) contents into the space between PM and
Sperm cannot fuse with membranes having fibrous mat of vitelline envelope proteins.
a positive resting potential, blocking fusion Contents of cortical granules
of sperm Proteases dissolve proteins that connect
Transient, because the membrane potential vitelline envelope proteins to the cell
can remain positive for only about a minute. membrane (e.g. ZP), and clip off sperms
Not efficient, as sperm bound to vitelline attached
envelope can still fertilise Mucopolysaccharides produce an osmotic
gradient that causes water to rush into the
space between the PM and the Vitelline
envelope, causing it to expand and become the
Fertilisation Envelope
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Peroxidase enzyme hardens the fertilisation

envelope by cross linking Tyrosine residues.
TOTI POTENCY
Totipotency is the ability of a single cell to divide and produce all the differentiated cells in an
organism
Spores are Zygotes are totipotent. They have the potential to develop into an embryo with all the
specialised cells that make up a living being. In case of zygote, cells are totipotent atleast till the 4-cell
stage embryo.
In some organisms, cells can de-differentiate and regain totipotency
As development proceeds, development potential of individual blastomeres gradually declines resulting
in pluripotent, multipotent, unipotent and terminally differentiated somatic cells.
Totipotency in mammals
Mammalian development commences with totipotent zygote which is capable of developing into all
specialised cells that make up the adult animal.
As development unfolds, cells of early embryo proliferate and differentiate into first 2 lineages
Pluripotent inner mass cell
Trophectoderm
Pluripotent cells can be isolated, adapted and propagated indefinitely in vitro in an undifferentiated
state as embryonic stem cells (ESCs). ESC can differentiate into cells representing the 3 major germ
layers: Endoderm, Mesoderm and Ectoderm.
Applications
Since many human diseases result from defects in a single cell type, pluripotent ESC are an unlimited
source of any cell or tissue type for replacement therapy.
They can be derived experimentally by nuclear reprogramming of somatic cells
Patient’s own somatic cells can be reprogrammed, eliminating any kind of immune reaction against
transplanted cells.
GASTRULATION
Gastrulation, is the process by which these presumptive areas of organ specific rudiments shift from
their original places to positions where these organs occur in adults.
e.g. Presumptive areas of mesodermal and endodermal organ rudiments move from the surface of
the blastula into the interior of the embryo
Gastrulation results in
Setting apart of the three primary germinal layers — ectoderm, endoderm and mesoderm, from
the blastoderm. Ectoderm is the outer tube, while Endoderm forms the alimentary canal and
organs derived from it.
Formation of primordial gut — Archentron
Types of Eggs
Microlecithal: Very small amount of yolk, young hatch quickly. Protochrodates and Eutherian
mammals
Mesolecithal: Intermediate amount of yolk, young hatch at a later stage of development. Lampreys
and Amphibians
Macrolecithal:Large amount of young, young hatch at a much later stage. Fishes, reptiles, birds
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Isolecithal: Distribution of yolk is even throughout the egg

Telolecithal: Yolk concentrated in one part of the egg. Leads to differentiation of vegetal and animal
poles
Types of Cleavage
Holoblastic Cleavage: Cleavage furrows pass through the entire egg. Equal cleavage in
Microlecithal eggs, Unequal cleavage in Mesolecithal eggs
Meroblastic Cleavage: Occurs in macrolecithal eggs. Cleavage takes place only in a disc at the
animal pole (blastodisc) and do not extend into the yolk
Major events during gastrulation
Rearrangement of cells due to morphogenetic movements of the cells of blastoderm

Three primary germ layers get differentiated
Cleavage rate is slowed down but cell divisions still take place
Growth in volume is insignificant
Oxidation becomes intensified — oxygen consumption increases and nature of metabolism changes
Nuclei become physiologically active and control activities of embryonic cells
Synthesis of new proteins
Movement of cells: Epithelial cells of the blastoderm show specific en-bloc movements during gastrulation.
Invagination: Infolding of cell sheet into embryo. e.g. Sea Urchin endoderm.
Involution: Inturning of cell sheet over the basal surface of an outer layer. e.g. Amphibian Mesoderm
Ingression: Migration of individual cells into the embryo. e.g. Sea Urchin mesoderm, Drosophila
neuroblasts
Delamination: Splitting or migration of one sheet into two sheets. e.g. Mammalian and bird hypoblast
formation
Epiboly: Expansion of one cell sheet over other cells. e.g. Ectoderm formation in amphibians, sea
urchins and tunicates
Gastrulation in Amphibians
Mitotic Divisions of zygote upto many celled embryo after 2 hours of fertilisation is Holoblastic.
1st cleavage: Meridional from animal to vegetal pole and divides zygote into 2 blastomeres.
2nd cleavage: Vertical at right angles to first cleavage. 4 blastomeres are formed

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3rd cleavage: Horizontal and a little above the equator — forms 4 micromeres and 4 macromeres
(differential cleavage)
16 to 64 cell stage: Morula
128 cell stage: Blastula (Triploblastic is a condition of Blastula where it has 3 germ layers)
Gastrulation converts the Blastula into spherical, bilaterally symmetrical, triploblastic Gastrula.
Gastrula undergoes Tubulation/neurulation to form Neurula
The objective of gastrulation in amphibians is to bring endoderm cells to the inside of the embryo, allow
ectoderm cells to coat the outside of the embryo and position mesoderm cells in between.
Onset: Marked by appearance of an infolding on the border-line of vegetal region and marginal zone
exactly below the gray crescent area. The infolding forms a narrow groove due to formation of bottle
cells.
Marks the beginning of Archentron and the opening represents the beginning of Blastopore. The fold
above the groove represents the dorsal lip of developing blastopore.
Involution
Cells of prospective chorda-mesoderm (mesenchyme cells present along the posterior margin of
grey-crescent) start rolling inward along the dorsal lip of blastopore into the blastocoels. The
chords-mesoderm cells will form the notochord.
Cells at the rim of blastopore undergo change in shape and this causes them to sink into the slit
Slit deepens and forms an archenteron. As it grows deeper, the Blastocoel shrinks. Archenteron
is lined by Endoderm.
Between the endoderm and ectoderm, the mesoderm spreads by involution.
Interior region of Archenteron gradually transforms into the pharyngeal region. It remains as the
foregut.
Epiboly
Active micromeres of the animal half that represent the future ectoderm, divide rapidly and
gradually spread over the macromeres of vegetal hemisphere and gradually push the blastopore
towards the vegetal pole.
Epiboly causes covering of gastrula by ectoderm
Invagination
Along with episodic growth of micromeres, macromeres of the presumptive endoderm of the
vegetal region stream inwards, increasing the archenteron. The insinking is known as
invagination.
During invagination, surface cells of the dorsal lip of blastopore becomes elongated and flash
shaped as they approach the blastopore. This process continues until bulk of each cell has moved
completely inside
Migration and involution of new micromeres from all other sides of the marginal zone towards and
around the blastopore lip. This results in formation of lateral and ventral lips of blastopore, which
finally assume a circular appearance
Circular blastopore gradually moves downward and gradually diminishes in size. In the end, it can be
seen as a small closed circle with a few macromeres jutting out of it in the form of yolk plug. Gradually
the yolk sac withdraws to the interior and blastopore gets reduced to a small slit.
Yolk Plug is the remaining patch of endodermal cells that is created during the formation of the
dorsal lip of the blastopore which remains exposed on the vegetal surface of the blastula that will
eventually be internalised by epiboly.
Fate of primary germ layers
Endoderm: Digestive and respiratory tracts and associated structures

Mesoderm: Skeleton, Circulatory system, muscles, excretory system, most of reproductive system
Ectoderm: Skin, sense organs and nervous system

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Gastrulation in Chick
Gastrulation in highly telolecithal eggs (large yolk situated near one end) is greatly modified, as
excessive yolk prevents cleavage in the vegetal pole region.
Yolky eggs of birds undergo discoidal mesoblastic cleavage. Cleavage occurs only in Blastodisc — a
small disc of cytoplasm 2-3 mm in diameter at the animal pole of the egg cell. Blastodisc is
differentiated in Area Pellucida and Area Opaca.
Area pellucida: Blastoderm is formed of an outer layer (epiblast) and an inner layer (hypoblast).
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Hypoblast is separated by underlying yolk by a subgerminal cavity.

Epiblast has mixed blastomeres for ectoderm, mesoderm and endoderm. Hypoblast represents
the presumptive endoderm only.
1st cleavage furrow appears centrally in the blastodisc.
Other cleavages follow to create a single layer blastoderm. The cleavages do not extend into the yolky
cytoplasm. So the early cleavage cells are continuous with each other with yolk at their bases.
Equatorial and vertical cleavages divide the blastoderm into a tissue 5 to 6 cell layers thick.
Gastrulation and embryo formation is confined to area pellucida only (embryonal area).
Closely related to formation of Primitive Streak whose long axis establishes the AP axis of the
embryo.
Gastrulation Events: Series of Morphogenetic movements of cells from Epiblast bring the presumptive areas
of blastoderm to their definitive positions.
Growth of blastoderm margins over the yolk to form yolk sac: These growth movements are unrelated
to the formative movements
Forward movements of hypoblast cells: Hypoblast cells from the posterior end of Area Pellucida flow
in a fountain like fashion and spread out towards anterior and lateral edges of the Blastodisc.
Formation of Primitive Streak:
6-7 hours after fertilisation, cells of lateral plate mesoderm begin to converge at the posterior
edge of Area Pellucida in the middle line.
The cell aggregate forms a thickening and marks the future caudal end of the embryo.
By the addition of more and more cells, the thickening becomes more demarcated and lengthens
in the cephalo-caudal axis, forming the Primitive Streak.
By 16h after incubation, the primitive streak is fully formed. Primitive streak defines the axes of
the embryo.
Primitive streak
Defines the axes of the embryo. Extends from P to A.

Migrating cells enter through its dorsal side and move to its ventral side.
It separates the left portion of the embryo from the right
Differentiation of Primitive Streak
Primitive Groove: Narrow median furrow extending along the middle line of the Primitive
streak.
Primitive Ridge: Thickened sides of the streak.
Hensen’s Node: Thickened cephalic end of the streak.
Morphogenetic and Gastrula Movements
Primitive streak formation: Movement of cells from the epiblast towards the posterior end of area
pellucida and their convergence in the middle line.
Primitive streak is a homolog of Blastopore. Movement of cells towards the thickened area is
comparable to episodic movements of cells towards amphibian blastopore.
Primitive streak has bipolarity: Anterior half comprises of elements of presumptive endoderm and
mesoderm both, while posterior half forms the extra embryonic mesoderm
Gastrula movements: Migration of cells of mesoderm and endoderm from the primitive streak. Cells
from the blastoderm detach, become amoeboid and move singly downward towards the hypoblast
(immigration). This is followed by arrival of new cells from epiblast to the primitive streak. Thus the
entire epiblast and primitive streak is a mass of moving cells
As a result of infiltration, the endoderm cells penetrate the hypoblast and push the hypoblast cells
outward and forward, forming the endoderm
Presumptive Notochord cells concentrate in the deeper parts of Hensen’s node and from there start
moving forward in the middle line as a dense rod like process — marks the beginning of Notochord.

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Presumptive mesoderm cells from primitive streak sink in and migrate laterally between epiblast and
hypoblast. These form the mesoblast. The mesoblast cells retain junctional contact with cells of
primitive streak.
At this stage, chick embryo consists of upper layer of ectoderm, middle layer of chorda mesoderm,
inner layer of endoderm. Presumptive notochords tissue underlies median embryonic axis with
mesoderm on either axis.
FATE MAPS
Fate Map is a topographical surface mapping of the blastula based on the ultimate fate of various part
of the blastoderm.
Observe which cell in the blastula develops into which organ in the embryo. During gastrulation, these
components migrate to their prospective positions. Thus, a fate map gives a clear understanding of
mechanism and outcome of gastrulation.
Artificial Markings
Vital Dye Marking

Devised by Vogt in 1925
Use vital dyes like Janus green, Nile Blue, Neutral Red, Bismarck Brown, etc. which colour the
embryo without killing it.
A block of agar is soaked in a vital dye and pressed against a chosen area of the blastoderm for a
short time. The dye diffuses from the agar into cells of the embryo.
Carbon Particle Marking
Stick carbon particle on surface and follow the movement in the embryo to construct the fate
map
Radioactive Marking
Label embryos with C-14, P-32 etc.
Natural Markings

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Certain eggs contain certain differently coloured areas. These areas can be observed and followed during
development.
Frog Egg
Animal hemisphere is black in colour — epidermis and nervous system
Vegetal hemisphere is yellowish in colour — endoderm of the gut
Gram crescent just below the equator — mesoderm and notochord
Egg of Ascidian
Animal pole consists of clear transparent cytoplasm, which gives rise to the ectoderm.
Vegetal pole is slaty grey cytoplasm, which develops into the endoderm.
Below the equator there is a pair of crescent like area on opposite sides — one area contains light
grey cytoplasm (develops into nervous system and notochord) and other area is yellowish
(develops into mesoderm)
Genetic Marking
Creating mosaic embryos having different genetic constituents (chimeric embryos)

Graft of quail cells into a chick embryo — chick and quail develop in a very similar manner, and a
graft of quail will participate in construction of various organs.
The chick that hatches will have quail cells in particular sites, depending on where the graft was
placed.
Fine structure maps of chick brain and skeletal system have been produced in this way.
NEURULATION
Occurs towards the end of Gastrulation. Transforms Gastrula into Neurula by establishing the CNS
Ectoderm gives rise to neural folds, flanking a neural groove along the axis from Blastopore towards
the future head
Folds meet mid-dorsally, forming neural tube. The anterior part becomes the brain, and the rest
becomes the Spinal Cord.
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Chordamesoderm (population of mesodermal cells) aggregates to form the notochord — It induces the
neural tube to form (Embryonic Induction). If the Chordamesoderm is experimentally removed,
notochord does not form
Process
Chordamesoderm that will go on to form the notochord induces neural plate formation — first step in
formation of neural tube
Ectoderm on the dorsal side of the embryo overlying the notochord thickens to form the neural plate.
At the neural fold stage, thickened ectoderm (neural plate) folds, leaving an elevated area along the
neural groove. Neural folds are wider in the anterior region, which goes on to form the brain.
During Neural tube stage, rural folds move closer together and fuse.
Neural groove becomes the cavity within the neural tube, which will later have the ability to transport
cerebrospinal fluid that aids in functioning of the CNS.
CELL LINEAGE
Cell Lineage of an organism is the pattern of cell divisions during its development
In lineage tracing, a single cell is marked in such a way that the mark is transmitted to the cell’s
progeny, resulting in a set of labeled clones.
Provides information about number of progeny of the founder cell, their location and their
differentiation status.
Applied to stem cell research and in modelling cellular heterogeneity in cancer.
Began with Whitman’s description of cleavage patterns in leech embryos. Later lineages were
described in nematodes, sea urchins and ascidians.
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Why Caenorhabditis elegans ?
Small, relatively simple and precisely structured organism. Anatomy has been described in detail and
one can map out the exact lineage of every cell in the body
Development is highly determinate, and cell-fate decisions are autonomous from surroundings
Number of somatic cells is highly limited. Makes the analysis straightforward and accurate
Fate of every cell in embryo has been determined using time lapse microscopy
Methods to Study Cell Lineage
Direct Observation
Direct observation or by reconstruction from fixed specimens.
Required embryos that were small, transparent and rapidly developing.
Nomarski Differential Interference Contrast Microscopy allows imaging of transparent
specimens.
Time Lapse microscopy in multiple focal planes has allowed entire cell lineages of individual
animals to be recorded digitally.
Clonal Analysis
In large, opaque or slowly developing embryos where direct observation of cell divisions is not
feasible
Necessary to mark individual cells by physical or genetic means, and later the progeny are
identified by expression of the marker.
Progeny of a single cell forms a clone
Cells can be labelled by injection with a non-diffusing dye such as fluorescein-conjugated
dextran.
Possible issue is that the dye gets progressively diluted with each division.
Used to study chick and mammalian neural development
Pattern of Cell Division
Proliferative: A — A + A
Cell divides symmetrically, each daughter behaves like its parent
Stem Cell Type: A — A + B
One daughter resembles parent and the other is of a different type
Diversifying: A — B + C
Daughters are different in fate from parent and from act other
Applications
Observe which cells in blastula develop into a cell of particular tissue or organ of the embryo. During
gastrulation, these cells migrate from their existing position to prospective position.
Understand Gastrulation
Classical Developmental Biology: Insights into development process of Caenorhabditis elegans and
Drosophila. These organisms display invariant patterns of cell division in which specification of cell
fates is correlated with cell division patterns. They reflect cell autonomous mechanisms of fate
determination and highly reproducible cell-cell interactions
Mutations in cell lineages help us understand mechanisms of cell fate specification and control of cell
proliferation
Stem cell research — provides information about how the cell behaves in context of intact tissue as
opposed to isolation, transplantation and in vitro culture.
Modelling heterogeneity in cancer
Can be performed without prior knowledge of what genes are expressed by cell of interest
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TYPES OF TISSUE GROWTH
Metaplasia
Changing of one cell type to another, usually as an adaptation to external environment.

External stimuli triggers altered gene expression, leading to differentiation of stem cells to a different
cell type.
Not a conversion of one differentiated cell type to another
Used most often in reference to epithelial cells, when basal cells (the stem cells of the epithelial layer)
switch from making one type of epithelial cell to another.
e.g. chronic irritation of cigarette smoke causing ciliated pseudostratified epithelium to be replaced by
squamous epithelium more able to withstand the insult)
Dysplasia
Increasing degree of disordered growth or maturation of the tissue (often thought to precede neoplasia)
Altered cell maturation, orientation and tissue architecture
Dysregulation of cell maturation and growth as a result of altered gene expression or genetic mutations
May progress to cancer or revert to original cells
e.g. Cervical dysplasia as a result of human papillomavirus infection.
Dysplasia is still a reversible process.
However, once the transformation to neoplasia has been made, the process is not reversible.
Neoplasia
"New Growth" - unregulated cell proliferation as a result of genetic changes.

Grow autonomously without any need for growth signals
Insensitive to normal growth-inhibitory signals
Capable of limitless replication, and – if they are malignant neoplastic cells – they invade vessels and
travel to different parts of the body causing cancer
Can have mutations in many different genes – e.g, the genes encoding growth factor receptors, signal-
transducing proteins, nuclear transcription factors, cyclins
DE-DIFFERENTIATION
Differentiation
During the early development of an organism, a certain cell type divides, and then its descendants
gradually form stable differences in morphology, structure, and function, finally producing numerous
different cell phenotypes.
This process of differentiation, is the fundamental basis for development.
During animal development, cells become progressively constrained to a certain cell type, and the
capacity to respond to a variety of differentiation signals declines after each generation. i.e. Process of
differentiation gradually reduces the potency of a cell.
In the final stage of differentiation, the cell is thought to stop dividing permanently.
De-Differentiation
Dedifferentiation is a process by which cells develop in reverse, from a more differentiated to a less
differentiated state
Differentiated cells revert to cells with an apparent stem cell or progenitor cell phenotype.
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These dedifferentiated cells redifferentiate into the new, differentiated cell phenotypes.
Can be observed at the levels of gene, protein, morphology, and function
Genetic Level: Cell undergoes reversion from a differentiated cell gene expression profile to a
progenitor cell gene expression profile. During the dedifferentiation process, development-
related gene activity is repressed, and genes that keep the cell in the undifferentiated state are
activated
Protein Level: Up-regulation of progenitor cell–related proteins and the down-regulation of
differentiated cell–related proteins
Morphological Level: Compared with mature cells, dedifferentiated cells are smaller, have fewer
organelles, and have a higher karyoplasmic ratio.
Functional level: Cell regains the capacity to proliferate, which means that a postmitotic cell can
reenter the cell cycle. Mature cells or lineage-committed cells might become multipotent or
pluripotent progenitor cells with the potential of differentiating into a variety of cell types
Different cell types follow different de-differentiation processes. Signalling pathways involved in de-
differentiation process include MAPK cascades, Wntß/catenin, Jak-STAT etc.
Examples of de-differentiation
Amphibians
Some amphibians have the ability to regrow damaged organs and lost body parts, such as a limb
or tail
This process involves the phenotypic reversion of fully differentiated cells, proceeding through
dedifferentiation, proliferation, and redifferentiation to form the required tissue or organ
Mammals (lesser degree of de-differentiation)
Germ Cells: High temperatures may induce de-differentiation. When monkey testes culture is
exposed to 43ºC, CK18 associated with immature Sertoli cells is induced
Nerve Cells: Following nerve injury, a differentiated, myelinating Schwann cell can
dedifferentiate, regain the potential to proliferate, and then re-differentiate during the repair
process
Renal Cells: Following injury, cells in the mature kidney begin to dedifferentiate, as evidenced
by the production of vimentin, a protein that is typically expressed in the mesenchymal cells that
give rise to the kidney cells
REGENERATION
Regeneration can be defined as the natural ability of living organisms to replace worn out parts, repair
or renew damaged or lost parts of the body, or to reconstitute the whole body from a small fragment
during the post embryonic life of an organism.
Regeneration is also a developmental process that involves growth, morphogenesis and differentiation.
Types of Regeneration
Physiological Regeneration: Replacement of cells caused due to wear and tear caused by day-to-day
activities.
Replacement of RBCs: Worn out RBCs are deposited in the spleen and new RBCs regularly
produced from the bone marrow cells. Life span of R.B.C's is only 120days.
Replacement of Epidermal Cells of the Skin
Reparative Regeneration: Replacement of lost parts or repair of damaged body organs. Wound is
repaired or closed by the expansion of the adjoining epidermis over the wound.
Regeneration of limbs in salamanders, lost tail in lizard
Healing of wound
Replacement of damaged cells.
Autotomy: Self-mutilation, where some part of the body is broken off on being threatened by a
predator

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Crabs break off their leg on approaching of the enemy

Holothurians throw off their internal viscera
Starfish breaks off an arm
Regenerative capacity in Animal Groups
The capacity of regeneration varies in its extent in various animal groups. Regenerative capacity is very
high among the protozoan, sponges and coelenterates.
Regeneration is faster in the young than in the adults.
Heteromorphosis: Regenerated part is not completely similar to the part lost
Invertebrates
Sponges: Entire body can be reconstructed from isolated body cells. The cells rearrange and reorganize
to form bilayered sponge body wall. Even 1/1000th part of the body regenerates into new organisms.
Hydra and Planaria: Small fragments of the body can give rise to a whole animal. When a hydra or a
planaria is cut into many pieces, each individual part regenerates into a whole individual
Annelids: Earthworms can regenerate some segments removed from the anterior and posterior ends of
the body.
Molluscs can regenerate only the eyes and heads while squids can also regenerate their arms.
Many arthropods (e.g., spiders, crustaceans, insect larvae, etc) can regenerate limbs only.
Echinoderms (like starfish, brittle star, sea lilly) exhibit autotomy. They can regenerate arms and parts
of the body.
Regeneration is an usual form of asexual reproduction in several lower groups of animals.
Vertebrates
Fishes: Lamprey can regenerate its lost tail. Some fishes have the ability to regenerate parts of its fins.
Amphibians: Can regenerate limbs, tail, external gills, jaws, parts of eye like lens and retina. Tail and
limb regeneration is found in the larval stages of frogs and toads.
Reptiles: Lizards exhibit autotomy. When threatened, the lizard detaches its tail near the base to
confuse its predator and later regenerates a new tail. The new tail differs from the old one in its shape,
absence of vertebrae and the kind of scales
Birds: Regeneration is restricted to parts of the beak.
Mammals: Regeneration is restricted to tissues only. External parts are not regenerated. Skin and
skeletal tissues possess great power of regeneration. The liver has the maximum capacity of

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regeneration. If one kidney is damaged or removed, the other enlarges to compensate the lost kidney
(compensatory hypertrophy)
Types of Regeneration based on Cellular Mechanism
Morphallaxis
Regeneration occurs mainly by the remodelling of existing tissues and the reestablishment of
boundaries, thus involving very little new growth.
Regenerated individual is much smaller initially. It subsequently increases its size and becomes
normal after feeding
e.g Regeneration of hydra from a small fragment of its body.
Epimorphosis
Regeneration involves dedifferentiation of adult structures in order to form an undifferentiated
mass of cells.
They are highly proliferating and accumulate under the epidermis, which has already expanded.
Within two days, bulge transforms into a conical hump.
Lump of dedifferentiated cells along with the epidermal covering is called regeneration bud or
regeneration blastema.
Dedifferential cells continue to proliferate and finally redifferentiate to form a rudiment of the
limb. The rudiment eventually transforms into a limb.
e.g. Limb regeneration in amphibians
Heteromorphosis
When a different organ develops from the one that has been removed
In shrimp Palinurus, eye is regenerated, If it is removed from the eye stalk. But if the eye is
removed along with optic ganglion, instead of eye an antenna like organ is regenerated.
This type of regeneration is exhibited by lower animals.
Super regeneration
The development of superfluous number of organs or parts of the body ( eg. Heads, tail, limbs) as
a result of regeneration is known as super regeneration.
When a deep incision is made on the head end of a planaria or earthworm, additional heads will
develop. Incisions in the middle part cause the development of both heads and tails.
Wolffian regeneration
Regeneration of a part of an organ from a tissue other than its original embryonic tissue is called
Wolffians regeneration.
In Newt, Triturus, if the lens of the eye is removed, a new lens is formed from uninjured iris.
The original lens is developed from epidermal ectoderm but the regenerating lens, formed from
iris is neurectodermal in origin
Mechanism of Regeneration
Regeneration is a complex process which involves histological and physiological events.
Salamander
Wound healing: The epidermal cells from the edges of the wound migrate and spread over the exposed
surface.
Blastema formation: A few days later, undifferentiated cells accumulate inside the epidermis, resulting
in a bulge (regeneration bud or blastema)
Redifferentiation and morphogenesis: Blastema develops rudiments of the lost organ, like the digits
which grow into new digits.
Growth: The regenerated limb increases and attains the size of a normal limb
Planarians and Hydra

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Undifferentiated cells called neoblasts which multiply and then migrate from the deeper parts of the
body to the cut surface.
Growth Factors: Wound healing is due to accelerated mitosis, mediated by proteins called growth
factors which act locally.
Epidermal growth factor stimulates the epithelium to undergo mitosis. EGF is also produced in the
salivary glands, which is why an animal's licking heals a wound.
Fibroblast growth factor stimulates the endothelial cells of the blood vessels to divide and heal the
injured blood vessels.
Platelet Derived growth factor which stimulates the mitosis of fibroblasts at the site of injury to fill in
the damaged areas under the blood clot.
Polarity in Regeneration: The body segments of Hydra or planarians exhibit distinct polarity during
regeneration. Their anterior end always regenerates into head and posterior into the tail
CELL TO CELL INTERACTIONS
During the process of animal development, cells of embryo interact with each other in a highly precise
manner, in the absence of which normal animal development is not possible. They affect growth,
differentiation and morphogenesis
Main types of cell to cell interaction are
1. Attachment based interaction — mainly control morphogenesis

2. Induction based interaction — mainly control growth and differentiation
Attachment Based Interaction
Differential Adhesion Hypothesis (Malcolm Steinberg, 1964): model to explain patterns of cell
sorting.
Each type of cell has a different set of proteins, and these differences are responsible for forming the
structure of the tissues and organs during development.
Cell actively move and organise to create tissue organisation
Cadherins and Cell adhesion
Cadherins are calcium dependent adhesion molecules

Integral membrane proteins of 723-748 amino acids, sharing common sequences.
Boundaries between tissues are created by
Different cell types having different types of cell adhesion molecules
Different cell types having different amounts of cell adhesion molecules
Cadherins are critical for
Establishing and maintaining inter cellular connections
Spatial segregation of cell types
Organisation of animal form
Mechanism of Cadherin action
Associated with actin bundles at the cytoplasmic domain through catenin proteins. Catenin is a hetero-
trimer consisting of α, ß and γ subunits
ß subunit is attached to cadherin, γ subunit attached to action microfilament, α subunit connect ß
subunit to the γ subunit.
Cadherins join cells together by binding to the same type of cadherin on another cell (homophobic
interactions, occur in the extracellular domain)
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Types of Cadherin
E-cadherin (Epithelial): Expressed on all early mammalian embryonic cells, even at the 1-cell stage.
Later, restricted to epithelial tissues of embryos and adults.
P-cadherin (Placental): Expressed mostly on trophoblast cells (placental cells of the mammalian
embryo that contact the uterine wall) and uterine wall epithelium. Possibly facilitates connection of
embryo to the uterus (P cadherin on uterine cells contact P cadherin on on trophoblast cells in mouse
embryos)
N-cadherin (Neural): First seen on mesodermal cells in the gastrulating embryo as they lose their E-
cadherin expression. Highly expressed on cells of the developing nervous system
EP-cadherin: Maintain adhesion between blastomeres of Xenopus blastula and is required for normal
movements of gastrulation.
Protocadherins: Calcium dependent adhesion proteins, but lack connections to cytoskeleton through
catenins. Important in separating Notochord from other mesodermal tissues during Xenopus
gastrulation.
Developmental contributions of attachment systems
Cadherin-Catenin complex forms the classic adherent junctions that connect epithelial cells together.
Since catenins bind the actin cytoskeleton of the cell, they integrate the epithelial cells together into a
mechanical unit.
In gastrula of frog, Neural tube expresses N-cadherin, while epidermis expresses E-cadherin. They
separate from each other such that neural tube is inside the body and epidermis covers the body.
Cadherins work in association with other adhesion systems
During implantation, trophoblasts contain both E and P cadherins and these recognise similar
cadherins on uterine cells.
Trophoblasts also have integrin which act as receptors for collagen and heparan sulphate
glycoproteins of the uterine wall.
Trophoblast cells have a modified Glycosyltransferase enzyme that extends out from the
membrane and that can bind to specific carbohydrate residues on uterine glycoproteins
Induction Based Interaction
Induction is a process in which one cell or a group of cells goes through specific developmental process
under the influence of another cell or group of cells. (Inducer and Responding Cell)
Induction is based on Paracrine, Autocrine and Juxtacrine signalling.
Autocrine interaction: Secretions by the cell affect the same cell
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Paracrine Interaction: Cell or tissue secretes proteins that induce changes in neighbouring cells
Juxtacrine Interaction: Inductions between cell membranes of adjacent cells or between a cell
membrane and an ECM secreted by another cell.
Responding cells must have the receptors signalling substances produced by the inducer cells.
Competent cells respond to inducing factors through signal transduction pathways
Reciprocal induction: Two interacting cells are both inducers and are competent to respond to each
other’s signals.
Cascades of inductive events are responsible for organ formation
Regionally specific induction can generate different structures from the same cells
Cross talk between signal transduction pathways allows the cell to respond to multiple inputs
simultaneously.
What processes are influenced? Division, Differentiation, Attachment, Migration, Cell death, de-
differentiation
Types of Induction
Instructive induction: Signal from inducing cell is necessary for initiating new gene expression in the
responding cell. e.g. When optic vesicle is experimentally placed under a new region of the head
ectoderm and causes that region of ectoderm to form a lens
Permissive induction: Responding tissue has all the potential, but only needs an environment that
allows the expression. e.g. many tissues need a solid substrate of fibronectin or laminin to develop. It
does not alter the type of cell that is to be produced, but only enables what has been determined to be
expressed.
Inducing Agents
Paracrine factors
Fibroblast Growth Factor Family
Hedgehog Family
Wingless Family
TGF-ß Family
Delta, Jagged, Serrate proteins in cell membranes activate neighbouring cells that contain Notch
protein in their membranes. Notch proteins are important in both vertebrate and insect nervous system
TGF-ß signalling
Required for processes like embryonic body plan, specifying and maintaining cell identities, cell
proliferation control, ECM production, cell death
Alteration of the pathway causes birth defects, disorders and diseases in humans
TGF pathways are highly conserved at molecular and functional levels. They act by activation of
specific target genes.

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Notch Delta Signalling
Lateral interactions between embryonic cells cause developmentally equivalent cells to assume
different fates. Notch pathway controls such lateral interactions
Notch is the receptor, Delta is the ligand. Both are large, transmembrane proteins.
Intra cellular domain of Notch is essential for transmitting a signal to the nucleus initiated by Notch-
Delta binding
Ligand induced activation — Notch undergoes a proteolytic cleavage — releases its intracellular
domain — Intra cellular domain translocates to the nucleus.
In the nucleus, it forms a complex with DNA binding protein Suppressor of Hairless SuH — stimulates
transcription — upregulation of expression of certain genes
Examples of Embryonic Induction
Neural Tube: If the ectoderm is removed, the neural tube does not form (but the embryo develops with
faulty CNS). When this removed ectoderm is cultured separately, it forms gastrula, but does not
undergo Neurulation. Thus, interaction of Ectoderm with layers underneath is important for formation
of Neural plate, neural folds and finally, the neural tube.
Lip of Blastopore: Cut a flap in Ectoderm at the lip of blastopore, remove the mesoderm and close the
flap. In absence of mesoderm, the ectoderm cannot undergo neural tube formation. Thus, mesoderm
has influence on ectoderm in its differentiation into nervous system
Dorsal Lip as Organsier: Cells from Dorsal Blastopore Lip is implanted on a second blastula, in a
position opposite to the original dorsal lip. Invagination occurs from both the positions in which there
are blastopore lips. Leads to formation of an extra set of body structures, showing the dorsal lip can
induce differentiation of an entire organism at the blastula stage.
Notochord induces Central Nervous System
Role of Cytoplasm in Development
Contains maternal mRNA and proteins — control cell functions during cleavage.

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Controls development till blastula or early gastrula stage in most animals except mammals.
During this cleavage period of development, cell division is so rapid, there is no time for
transcription to occur and embryonic genome is not expressed. (Some animals can develop to
blastula level even if nucleus is removed. But mammalian cleavage divisions are slow, so even in
early stages zygotic genome is transcribed)
If transcription is blocked, the cleavage divisions can still continue. But if translation is blocked,
cleavage is stopped — shows that mRNAs vital for cleavage divisions are already present in the
egg
Grey Crescent: When a sperm penetrates the egg, the cortical cytoplasm shifts revealing the grey
crescent. Cytoplasmic factors within the grey crescent are vital for blastopore formation. It also forms
the notochord after moving inside the embryo during gastrulation and is responsible for induction of
CNS.
Spemann’s Experiment
Gradients in the egg cytoplasm (Bicoid, Hunchback, Caudal, Nanos) determine AP polarity later on in
the embryo
Cytoplasm contains cytosolic receptors for signal molecules, that direct pattern of differentiation
through transduction pathways
BODY AXES
Overall body plan of organisms id defined by three major axes
Anterior Posterior (AP)

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Dorso Ventral (DV)

Left-Right (LR)
Axes are established in development via the expression of specific sets of genes that regulate which cells will
develop into specific structures.
DROSOPHILA A-P AXIS
AP polarity of embryo, larva and adult has its origin in AP polarity of the egg.
Maternal effect genes expressed in mother’s ovaries produce mRNAs that are placed in different
regions of the egg.
These mRNAs encode transcriptional and translational regulatory proteins that diffuse through
syncytial blastoderm and activate or repress the expression of certain zygotic genes.
Bicoid and Hunchback mRNAs: protein products are critical for head and thorax formation
Nanos and Caudal mRNAs: protein products are critical for the formation of the abdominal segments
Mechanism
Hunchback and Caudal mRNA are distributed throughout the oocyte.
Upon fertilisation, Bicoid protein forms the highest gradient at the Anterior, Nanos at the
posterior
Bicoid is present in anterior portion of unfertilised egg, tethered to anterior microtubules
Nanos is bound to cytoskeleton in the posterior position.
Bicoid inhibits translation of Caudal, which is expressed only in posterior.
Nanos in conjunction with Pumilio protein binds to hunchback, preventing its transcription.
Hunchback is thus expressed only in anterior
Bicoid elevates level of Hunchback by binding to enhancers of hunchback.
Bicoid and Hunchback — Anterior
Nanos and Caudal — Posterior
FROG D-V AXIS
Vertebrate axes do not form from localised determinants in the various blastomeres
Arise progressively through a sequence of interactions between neighbouring cells
Regulative Development: Cell’s fate is determined by interactions between neighbouring cells
(induction)
Process of DV axis establishment

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Dramatic movements in the cytoplasm following the fertilisation of amphibian eggs, and in some
amphibians these movements expose a gray, crescent shaped area of cytoplasm (grey crescent) in the
region directly opposite to the point of sperm entry.
First cleavage plane splits the grey crescent equally into two blastomeres.
Blastomeres in the grey crescent have rich presence of Dishevelled (Dsh) protein due to the
cytoplasmic movements of fertilisation
Dsh stabilises ß catenin protein. The ß catenin acts as anchor for cell membrane cadherins or as a
nuclear transcription factor. It begins to accumulate in the dorsal region of the egg, due to presence of
Dsh protein.
It continues to accumulate preferentially in the dorsal side throughout early cleavage, and this
accumulation is seen in nuclei of dorsal cells
Binds to a ubiquitous transcription factor TCF3. TCF3/ß catenin complex binds to promoters of several
genes whose activity is critical for dorsal axis formation.
e.g. Siamois gene. In absence of ß catenin, TCF3 inhibits this gene. But TCF3/ß catenin complex
activates this gene.
Siamois protein is critical for expression of organiser specific genes.
It binds to promoter of goosecoid gene and activates its expression.
Goosecoid protein activates many genes related to dorsal specification
HOMEOTIC GENES
Development related genes in almost all animal groups.

During early embryonic development, these genes control segment identity. After this, proper number
and placement of embryonic segment structures occurs (legs, antennae, eyes)
Discovered in 1940s by Edward Lewis in Drosophila
Mutations in these genes cause body segments to lose their individual identity and become like some
other segment. e.g. Mutations in the Antennapedia gene cause legs to develop on the head of a
Drosophila in place of the antenna.
Homeosis: Abnormal developmental phenomenon.
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They are expressed early in the embryo and are activated by specific concentrations of gap, pair rule
and segment polarity genes. Products of Homeotic genes activate other genes that encode these
segment-specific characteristics.
Homeotic Genes in Drosophila
First identified in Drosophila, which has 8 Homeotic genes.

These genes are arranged in two clusters on chromosome 3. Each cluster is called a Homeotic Complex
(Hom-C)
Antennapedia Complex
Labial — head segments
Antennapedia — thoracic segments
Sex Combs Reduced — thoracic segments
Deformed — head segments
Proboscipedia — labial paps in adults
Bithorax Complex
Ultra bithorax — identity of third thoracic segment
Abdominal A and B — identity of abdominal segment
Order of genes in the HOM-C is the same order in which genes are expressed along the AP axis of the
body
Other species
Homologs of Drosophila homeotic genes have been found in almost every animal species
They are arranged similar to the insect HOX complex — suggests that all Homeotic gene complexes
originated by duplication of a single primordial complex and they have preserved the basic
organisation
Mouse/humans: 4 such complexes — HoxA, B, C and D. Each complex is on a different chromosome.
Action of Homeotic Genes
In Drosophila, homeotic genes act after body segments have been laid down and each segment has also
established its polarity.
They provide developmental identity to individual segments by activating specific realisator genes,
which control organ development.
Homeotic genes encode TFs. Each gene has a subset of 180 nucleotides that encodes a 60 amino acid
long helix-turn-helix type of DNA binding domain.
GENES IN CHICK DEVELOPMENT
Regulatory Genes: Transcription Factors, Components of Signal Transduction (including signal

substances and signal receptors).
Effector Genes: encode enzymes and structural proteins of specific tissues or cells which can directly
lead to differentiation
There are significant and fundamental similarities between human and chicken. Differences are present
mainly in the regulatory elements.
Important Genes

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Vg1 and Wnt8c: Forms signalling factors during early development which leads to initiation of
gastrulation
Chick en-2: Needed for proper Neurulation in chick. Encodes a TF
BMP gene product is a signalling factor and it induces ventral development in chick.
Cells of Hensen node and its derivatives act like organiser — secrete Chordin, Noggin and Nodal —
antagonise BMP and dorsalise the ectoderm and mesoderm
FGFs generate neuronal phenotypes in the epiblast cells. Produced in Hensen’s node and the Primitive
streak.
LAMA3: encodes Alpha 3 subunit of Laminin S, a component of the basal membrane of the skin which
plays a role in dermal differentiation
Tinman gene: Encodes a TF necessary in development of heart
Nodal (paracrine signalling factor) and PITX2 (TF): Distinction between right and left sides
Sonic Hedgehog: Separation of single eye field into two bilateral fields. If this gene is mutated, it
results in Cyclopia — a single eye in centre of head
Transforming Growth Factor (TGF): Development of Epidermis. Made by the basal cells and
stimulates their own division (Autocrine)
GENES IN HUMAN DEVELOPMENT
Developmental processes in animals are definitive, heritable character of each species in which body
plans are morphological outcomes.
Diverse developmental processes in bilateria are generated with similar sets of regulatory and
signalling genes.
Genomic instructions are either in form of regulatory or effector genes
HGP: 4000 genes in humans are related to the developmental process.
Secreted Signalling Proteins
SHH: Sonic Hedgehog: Patterning neural tube, gut and limb

EDN3: Endothelin: Regulates Vasoconstriction, required during development for differentiation of
neural crest derivatives
GH1: Growth Hormone: Polypeptide Hormone expressed in Pituitary gland, responsible for regulating
growth
LEFTB: Lefty2: Nodal related signalling protein expressed on left hand side, helps to establish LR
symmetry
Receptors
FGFR3: Expressed strongly in cartilage and nervous systems. Plays a role in bone development.
EDNRB: Endothelin B receptor: found on neural crest cells and required for their differentiation into
melanocytes and enteric ganglia in the gut
KIT: Receptor Tyrosine Kinase: Development of blood cell lineage, melanocytes and germ cells.
GHR: Growth Hormone Receptor: Transduces signals from Growth Hormone
Transcription Factors
HOXD13: Involved in pattern formation, confers positional information along craniotomy-caudal axis
and in limb development.
PAX6: TF with multiple roles in eye development
TBX5: Expressed specifically in forelimb region of developing embryo
SRY: Expressed in gonadal ridge of male embryos, essential for male sexual differentiation, found on
Y chromosome

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Structural Proteins
COL6A1: Component of microfibrils in the ECM, provides structural rigidity

ELN: Elastin: Protein of ECM that allows tissues to return to their original shape after deformation
LAMA3: Component of basal membrane of skin, which plays an important role in Keratinocyte
differentiation
USH2A: ECM protein required for eye and inner ear development
Enzymes
WRN: Helicase involved in repair of dsDNA breaks

CYP11B1: Synthesis of aldosterone, which acts on kidneys and regulates mineral and water balance
EXT1: Glycosyltransferase required for synthesis of Heparan Sulfate (constituent of ECM)
METAMORPHOSIS
Metamorphosis is the development of an adult from the larval form

In many species, early embryonic development leads to a larval form which is very different from the
adult form.
Larval forms are specialised for some function like growth or dispersal.
During metamorphosis, developmental process are reactivated by specific hormones. The entire
organism changes extensively through dramatic developmental change.
Metamorphosis in Amphibians
Metamorphosis changes an aquatic organism for a primary terrestrial existence.

Urodeles: Resorption of tail fin, destruction of external gills, change in skin structure
Anurans: Changes are much more dramatic, with almost every organ undergoing some transformation.
Aquatic locomotion with tail fins to Terrestrial tetrapod
Respiration: Gills, skin and Lungs to Skin and lungs. Haemoglobin structure also undergoes a
change (Adult haemoglobin binds oxygen more slowly and releases it more rapidly)
Diet: herbivore to carnivore, small mouth with horny jaws to large mouth with long tongue
Excretion: Largely ammonia to largely Urea (Urea excretion requires less water). Liver begins to
synthesise urea cycle enzymes
Integument: Thin, bilayered epidermis to Stratified squamous epidermis, thin dermis to
developed dermis, mucus glands develop in dermis
Lateral line system is lost, Tympanic membrane develops
Porphyropsin as major retinal pigment (like in freshwater fishes) to Rhodopsin (like in terrestrial
and marine vertebrates)
Role of Thyroid Hormones
Metamorphic changes in frog development are brought about by secretion of Thyroxine (T4) and
Triiodothyronine (T3) from the Thyroid during metamorphosis.
T3 causes metamorphosis at much lower concentrations than T4.
Thyroid destroyed: No metamorphosis, become giant tadpoles
Add thyroid extract: Early/premature metamorphosis
Tail degeneration and Limb formation are both initiated by Thyroid hormones (same stimulus can
cause some tissues to degenerate and others to develop and differentiate)
Organ Morphological Changes Biochemical Changes

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Brain Restructuring, Axon guidance and growth Cell division, Apoptosis, Protein
synthesis
Induction of albumin and urea cycle

Liver Functional differentiation, restructuring
enzymes, larval adult haemoglobin switch
Re-position, New retinal neutrons, altered

Eye Porphyropsin to Rhodopsin switch
lens
Skin Keratinisation, granular gland formation Induction of collagen, keratin
Morphogenesis of bone, skin, muscle,

Limb bud, lung Cell proliferation, gene expression
nerve etc.
Programmed cell death, induction of Lytic

Tail, Gills Total tissue regression and removal
enzymes
New structural and functional

Intestine Major remodelling of tissue
constituents
Immune
Redistribution of immune cell populations New immune competence
System
Muscle Growth, differentiation, Apoptosis Induction of Myosin heavy chain
Mechanism of Thyroxine Action
Largely at the level of transcription, activating transcription of some genes and repressing transcription
of others.
Transription of genes for Albumin, Adult Globin, Adult skin keratin etc are activated by Thyroid
hormones
About 20 Thyroid hormone regulated genes participate in intestinal remodelling.
T3 —> Transcriptional activation of Thyroid receptor genes (part of steroid hormone receptor
superfamily)
2 major types of T3 receptors — TR α and TR ß
mRNA and proteins of both TR are present in relatively low levels in pre-metamorphosis
tadpoles, Increase before Thyroid hormone is released
TR bind to specific sites on Chromatin with help of Retinoic Acid Receptors, even before
Thyroid hormones are present. When T3/T4 is not present, it represses gene transcription.
When T3/T4 enters the cell and binds to chromatin-bound receptors, the hormone receptor
complex acts as a transcriptional activator.
Synthesis of TRs accelerates, coinciding with Metamorphosis
More T3 receptors, means the tissue is more competent to respond to small amounts of T3.
Metamorphic climax is brought about by enhanced production and induction of more T3
receptors.

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Regulation of Thyroxine Action
Prolactin inhibits Thyroid hormone action: If up regulation of TR is experimentally blocked by

Prolactin, the tail is not resorbed, adult specific keratin gene is not activated.
Injection of Prolactin stimulates larval growth and inhibits metamorphosis
Not confirmed if that is the natural role of Prolactin
Metamorphosis in Insects
Transformation of an immature insect from larvae to sexually mature adult, involving morphology,
function and habitat changes
Hemimetabolous Metamorphosis: Immature stages called Nymph that resemble adults. Only changes
are increase in size and development of sexual organs and wings. e.g. Dragonflies, Damselflies etc.
Holometabolous Metamorphosis: egg — larvae — pupa — adult
Ametabolous Metamorphosis: Immatures are almost the same shape as adults. No distinct
rearrangements of body structures between immature and adult.
Instar: Developmental stage of arthropods, between each bolt until sexual maturity is reached.

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Events of Metamorphosis
At the time of early development, in the developing egg, the cells are segregated into two groups —
one group for working at the larval life and the second group to take charge during pupal and adult life
In a growing larva, the larval cells increase only in size but never undergo division.
The second group of cells remain inactive in the body of larva. When the larva is full-grown, second
group of cells take over the charge.
During metamorphosis, most of the larval organs in the pupa except the central nervous system and
developing reproductive organs are broken down by enzymes (Histolysis)
Moulting and metamorphosis in insects are controlled by hormones
Hormonal Control
Brain
In the brain there are four groups of neurosecretory cells
Secrete Prothoracotropic (PTTH) or brain hormone that activates the prothoracic glands which in
turn produce moulting hormone
Protocerebrum sends the neurosecretory axons to the corpora cardiaca, a pair of small glands
which lie posterior to the brain.
Prothoracic Glands (Moult gland or Ecdysial Gland)
Mass of glandular tissue of non- nervous origin
Produces a steroid hormone called Ecdysone.
Stimulates growth and initiates the process of moulting and shedding the old cuticle of the larva
and the new cuticle is formed beneath the old cuticle
Corpora Allata
Paired non-nervous secretory cells and situated behind the brain posterior to corpora cardiaca
Corpora allata secrete fat soluble hormone, called juvenile hormone (JH) - chemcially related to
ecdysone, also a steroid
Keeps the larval cells active and also controls the qualitative changes in the body during
metamorphosis
As long as the juvenile hormone is secreted from the corpora allata, the pupa and imago (adult)
stages are not developed.
After a certain period the production of ecdysone instructs to stop the flow of juvenile hormone
and triggers the imaginal buds to be active
Absence of juvenile hormone causes the death of larval cells and they are used as nutrients for
the growing imaginal buds.
If the amount of juvenile hormone (JH) becomes lower in the blood, the moulting from larva to
pupa takes place and absence of juvenile hormone in the blood, there occurs from pupa to adult
moult.
BLASTOGENESIS
In asexual reproduction, new individual develops from a group of cells called Blastema.
Development that starts from Blastema is called Blastogenesis.
Individuals resulting from asexual reproduction are referred to as Blastozooids.
Different modes of Blastogenesis
Fragmentation
Spontaneous fragmentation of the body and regeneration of the missing parts — sponges, sea
anemones, planarians
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Tips of branches of some sponges break regularly and produce masses of cells that become
young sponges.
Fission
Protozoans, Cnidarians, Annelids
New individual is formed from a relatively large portion of the body of the maternal organism,
and differentiated organs and tissues or their parts are passed on to the offspring.
Multiple Fission in some Protozoa — repeated nuclear division without Cytokinesis to form a
multi nucleate organism. Cytoplasm later cleaves to form many uninucleate individuals
In some corals (Anthozoa), fission occurs in form of longitudinal fission, with the division plane
in the long axis of the body.
Budding
Sponges, Cnidarians, Cestodes, Annelids and Tunicates
New individual develops from a small outgrowth on the surface of the parent.
Organs are not passed on to the bud, but it is supported by parental organism in initial stages of
development
Gemmule and Statoblast Formation
Freshwater Sponges and Bryozoans
Highly specialised asexual reproductive bodies called Gemmules in sponges and statoblasts in
Bryozoans.
New individuals develop from groups of cells which become completely cut off from the
maternal individual, so that development is independent of the maternal body
Formed in interior of the parental body from a number of undifferentiated cells which become
enclosed in a shell with spicules (sponges) or chitinous envelope (Bryozoans)
On destruction of parental body, they are set free, and when environmental conditions are
favourable, the cells burst out and give rise to new individuals
Demospongia produce gemmules with flagellated epithelium which settle after a free swimming
period and give rise to new individuals
Evolutionary Significance
Only one parent with no special reproductive organs or cells. Either male or female can produce
identical copies as soon as it becomes an adult. Simple, direct and rapid mode of reproduction.
However, the offspring are clones of parents unless mutation is introduced
Mutation is expressed immediately and thus, evolution proceeds very quickly. In sexual reproduction,
mutations are often recessive and hence masked by the other parent at a homologous locus.
Better resistance to adverse environmental conditions. Can adopt different modes of asexual
reproduction, depending on factors
TERATOGENESIS
Teratogenesis is a prenatal toxicity characterised by structural or functional defects in the developing

embryo or foetus.
Includes intra uterine growth retardation, death of embryo, transplacental carcinogenesis
Many cases, due to teratogenesis, babies born have a recognisable malformation
Malformations: Congenital abnormalities caused by genetic events (mutations, aneuploidies,
translocations). e.g. Down’s Syndrome
Teratogenesis: Abnormalities caused by exogenous agents — chemicals, viruses, radiation,
hyperthermia
Induced Teratogenesis
Teratogens
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Drugs and Chemicals: Alcohol, Cigarette Smoke, Cocaine, Cortisone, Lead, Tetracycline,
Warfarin, Bromine
Ionising Radiation
Hyperthermia
Micro organisms: Parvovirus, Rubella, Toxoplasma
Metabolic Conditions in Mother: Rh incompatibility, Diabetes, Phenylketonuria
Different Teratogens have different critical time windows of highest susceptibility. The time from
period of Day 15 to Day 60 of gestation is critical for development of many organs, and during this
time many teratogens have worst effect.
Important in forming AP axis of mammalian embryo and in forming the limbs

But in large amounts it acts as a teratogen
Retinoic Acid RA binds to RA receptor and can control expression of HOX genes.
Can affect the AP axis, and inhibits neural crest migration from cranial region
of the neural tube
Consumption of alcohol by pregnant women leads to

Impairs Neural Crest migration
Apoptosis of neurons by superoxide radicals that can oxidise cell
membranes and lead to Cytolysis. Activation of GABA receptors and
Alcohol
simultaneous inhibition of Glutamate receptors also leads to apoptosis.
Interference in cell adhesion
Foetal Alcohol Syndrome: Small head size, narrow upper lip, smaller brain,
defects in neuronal and glial migration
Interfere with normal functions of hormones
Mimic the effects of natural hormones by binding to their receptors.

Block binding of hormone to its receptor
Endocrine
Block synthesis of hormone
Disruptors
Interferes with transport of a hormone or its elimination from the body. Rats
exposed to Polychlorinatedbiphenyl pollutants have low levels of Thyroid
hormones.
Decreases levels of PAX1 gene transcription

Valproic Acid
Major and minor bone defects
Jervine and Inhibit cholesterol synthesis and prevent Sonic Hedgehog signalling.
Cyclopamine Causes Neurological damage
Nicotine Infants are smaller
1/6 chance of giving birth to infant with eye cataracts, heart malformations and
Rubella
deafness

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Cytomegalovirus Infection of early embryos is almost always fatal

Infection of later embryos leads to blindness, deafness, cerebral palsy and
mental retardation
Ionising Radiation Break chromosomes, alter DNA structure
Fevers Heat increases illegitimate DNA recombination in cells
Genetic Teratogenesis
Phenomenon of severe deviation from normal developmental pattern due to genetic factors or pre natal
events
Major contributor to congenital malformations (physical defect present in a baby at birth)
Development of a structure is arrested, delayed or misdirected early in embryonic life and the effect is
permanent.
20-25% of perinatal deaths
Nowadays, they can be diagnosed early in pregnancy
Causes
Single Gene Malformation:

Result of a single mutated gene
Aniridia — absence of Iris, occurs due to mutation in gene PAX6
Autosomal Dominant Malformation
50% chance that the child will inherit a disease, if either parent has the disease
100% if both parents have it
Achondroplasia — leading to severe skeletal disorder that causes dwarfism
Marfan syndrome — Mutation in FBN1 which encodes connective protein Fibrillin. Patients are
tall, with long limbs and long thin fingers
Autosomal Recessive
Two copies of the gene must be mutated. Affected person typically has both parents as
unaffected carriers.
Spinal Muscular Atrophy — degeneration of motor neurons
X Linked
Mutations in genes on X chromosome
Manifests in all males with the mutation, but in females only if the other X chromosome has the
defect
Muscular Dystrophy
Aneuploidy
Numerical aberration affecting one or few chromosomes.
Down syndrome/Trisomy X — mental retardation and multiple malformations
MORPHOGENESIS

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Morphogenesis is the shaping of the multicellular body and its organs. It results from pattern
formation, which is the organisation of differentiated tissues into specific structures.
It is concerned with shapes of tissues, organs and entire organisms based on positions of various
specialised cell types.
Three fundamental aspects of animal development: Growth, Differentiation, Morphogenesis
Process
Gastrulation: Beginning stage. Patterning in Embryo

Organogenesis: All the organs are formed at their right places, in right form and polarity and in right
numbers.
Four main aspects
Tissue formation from a group of cells
Organ formation by tissues
Proper cell migration and formation of structures at right places
Correct location, pattern, polarity and number of organs
Cell arrangement types in the embryo
Epithelial Cell: Tightly connected to one another in sheets or tubes
Mesenchymal Cell: Unconnected to one another, and operate as independent units.
Processes involved in cell arrangements
Differential cell-cell attachment through controls on cell adhesion molecules
Direction and number of cell divisions, through morphogenetic inductions controlling cell
division cycle
Cell death through morphogenetic inductions controlling Apoptosis
Cell movement through control on cell adhesion molecules
Cell shape changes through morphogenetic inductions controlling cytoskeletal element
organisation.
Cell growth through morphogenetic inductions controlling cell growth
Changes in composition of cell membrane or surrounding matrix.
Morphogenetic processes displayed by Mesenchymal cells
Condensation — Mesenchyme to Epithelium — Cartilage Mesenchyme
Cell Division — Mitosis to produce more cells — Limb Mesenchyme
Cell Death — Cells die — Interdigital Mesenchyme
Migration — Cells move at particular times and places — Heart Mesenchyme
Matrix Secretion and Degradation — Synthesis or removal of extra cellular layer — Cartilage
Mesenchyme
Growth — Cells get larger — Fat Cells
Morphogenetic processes displayed by Epithelial cells
Dispersal — Epithelium to Mesenchyme — Mullerian duct degeneration
Delamination — Epithelium to Mesenchyme (part of structure) — Chick hypoblast
Shape change and growth — Cells remain attached as morphology is altered — Neurulation
Cell Migration — Rows of epithelia merge to form fewer rows — Vertebrate gastrulation
Cell Division — Mitosis within row or other direction Vertebrate gastrulation
Matrix secretion and degradation — Synthesis or removal of extra cellular layer — Vertebrate
organ formation
Migration — Formation of free edges — Chick ectoderm
Specific movements during gastrulation
Delamination: Splitting or migration of one sheet into two sheets — mammalian and bird
hypoblast formation
Epiboly: Expansion of one cell sheet over other cells — Ectoderm formation in amphibians, sea
urchins and tunicates
Invagination: Infolding of cell sheet into Embryo — Sea urchin endoderm
Involution: Inturning of cell sheet over the basal surface of an outer layer — Amphibian
mesoderm
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Ingression: Migration of individual cells into the embryo — Sea urchin mesoderm, Drosophila
neuroblasts
Morphogens
Morphogens are molecules which control development of specific structures during embryonic
development
They are mostly small globular proteins — signalling factors or transcription
Essential for normal structure formation. Absence or malfunction leads to malformations, cancers etc.
Discovered and aimed by Lewis Wolpert in Drosophila
Genetic Control
Regulated activities of genes and their products, which depend on morphogens

Transcription Factors: Hunchback, Caudal, Bicoid, Nanos etc. Necessary for inducing or maintaining
the expression of different target genes at distinct concentration thresholds.
Signalling Molecules: Most morphogens are in this category. Induce specific pattern of gene expression
through signal transduction. e.g. TGF, Hedgehog
Environmental Effects
Teratogens during morphogenesis
Mechanism of action
Two classical theories

Extracellular diffusion theory: Suggested that morphogens act on cells from the surface. e.g. Dpp
morphogen
Transcytosis theory: Cells transfer Morphogen molecules through endocytosis.
Modern Concept — 2 classes of morphogens
Transcription factors: Hunchback, Caudal, Bicoid, Nanos etc. Necessary for inducing or
maintaining the expression of different target genes at distinct concentration thresholds.
Signalling Molecules
Paracrine Factors: 4 major families — fibroblast growth factor (FGF), Hedgehog Family,
Wingless Family, TGF ß
Juxtacrine Factors: Delta, Jagged, Serrate proteins. Important receptors in nervous system
NEOTENY
Neoteny or Juvenilisation, is the retention of juvenile traits by adults in a species. It has two forms depending
upon the capability of individuals to breed
Paedomorphosis Paedogenesis
Adults retain traits that are seen only in Reproduction by an organism that has not
juveniles. achieved physical maturity
Proposed by Walter Garstang in 1922.

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In many domesticated animals, human directed Production of offspring by the larval or

breeding has possible led to Paedomorphosis juvenile form and elimination of the adult
with selection for juvenile characters like phase of the lifecycle.
docility Found in insects in which the larval stage
reproduces without reaching maturity.
Flightless Birds
Ostriches, Emu, Cassowaries, Kiwis are believed to have evolved by retaining characters of chicks and
losing the ability to fly
Physical proportions of these flightless birds resemble those of chicks of flying birds
Humans
Sparse body hairs and enlarged heads are similar to baby primates
Lactose tolerance in adults
Females do not acquire toughened skin, coarse body hair, thyroid cartilage. Paedomorphic characters in
women are considered desirable
Juvenile chimpanzees have a near identical bone structure to humans.
Loss of pigmentation in skin, eyes and hair
Form of the external ear
Central position of Foramen magnum. It migrates backwards during ontogeny of primates
High relative brain weight
Labia major of women
Form of the pelvis
Structure of hand and foot
Absence of eye brow ridges
Small teeth and variation of tooth row and cranial structures.
Thin skull bones
No rotation of big toe
Prolonged period of growth and infantile dependency
Crying to attract sympathy
Absence of cranial crests
Origin of Chordates
Nearest relatives of Chordates are Tunicates (marine filter feeders)

Tunicates are sessile in their adult form, but have a motile larval form
The larva has a notochord similar to that found in chordates
At some point the motile larvae of the tunicates became sexually mature before metamorphosis and
gave rise to free swimming chordates.
Neoteny in Amphibia
Natural Paedomorphosis occurs in many species of amphibians, especially Ambystomatid and Protean
salamanders.
Neotenes retain ability to regenerate limbs, tails and nearly every organ in the body.
Types of Neoteny

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Obligate Neoteny
Never fully metamorphose and retain larval characteristics to varying degrees into adulthood.
Insensitive to Thyroid hormone doses.
E.g. Amphiumidae, Sirenidae, Cryptobranchidae
Inducible Obligate Neoteny
Metamorphosis can be induced by manipulating the Thyroid function in the laboratory.
e.g. Ambystoma mexicanum
Facultative Neoteny
Occurs in individuals or entire populations as a result of environmental factors.
In extremely cold temperatures where terrestrial existence is inhospitable, individuals or
populations may remain aquatic, retaining larval characters into adulthood.
e.g. Salamandridae, Ambystomatidae
Axolotl
Sexually mature at 18-24 months

Features typical of salamander larvae — external gills, caudal fin (from behind the head to the vent)
Heads are wide and the eyes are lidless
Limbs are underdeveloped with long, thin digits.
Barely visible vestigial teeth, which would have developed during metamorphosis
Three pairs of external gills are used for respiration.
Buccal pumping (gulping air from the surface) can also be used to provide oxygen to lungs.
Physiological Basis
Thyroid Stimulating Hormone (TSH) — Thyroid — Thyroxine — metamorphosis

Hypothalamus fails to produce hormone that cause Pituitary to produce TSH
Axolotl can be induced to metamorphose by injecting Iodine or Thyroxine hormone
Artificial metamorphosis is largely unsuccessful as there is a strong genetic basis for Neoteny
CELL DEATH
Cell Death is a condition when the cell irreversibly loses

Chemical organisation
Physiological functions
Ability to reproduce
There is progressive impairment of the cell’s normal function and structure prior to irreversible loss of
cellular vitality.
Types of Cell death
Necrosis: Cell death due to injury. Always pathologic

Apoptosis: Programmed cell death. Part of normal development and defence mechanisms in which
unwanted cells are removed from the animal body. The cell dies through activation of an internally
controlled suicide program.
APOPTOSIS
In development of multicellular processes cells must die during processes like embryogenesis,
metamorphosis, tissue turnover.
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Blebbing: Formation and pinching off of vesicles from the outer surface of the cell.
Membranes of organelles such as mitochondria are disrupted. Their contents leak into the Cytoplasm.
Heteropycnosis: Nuclear and Chromatin condensation
Cleavage of DNA into characteristic 180 bp fragments
Cell loses normal shape and breaks into small cell fragments
Formation of Apoptotic bodies which are engulfed and degraded by neighbouring cells.
Instances of Apoptosis
In C. elegans cells will die predictably at a defined time and place in each animal (somatic cell lineages
have been completely defined)
During vertebrate development, in immune and nervous system
Elimination of surplus cells by apoptosis is required for proper connection in the brain (synaptic
connections). Mice have malformed cerebrum if Apoptosis is inhibited.
Apoptosis of interdigital tissues during embryonic development to form fingers and toes. This process
distinguishes the chicken’s foot from the webbed foot of a duck
Resorption of tadpole tail
Sloughing off of inner line of uterus at the start of menstruation
Liquidation of insect muscles cells during metamorphosis
Physiological regulation of tissues like Placenta
Pathological responses to mutagens, toxins and hormones.
Process
Apoptosis involves activation of a pathway that leads to suicide of the cell. It is an active process that
depends on RNA and protein synthesis by the dying cell.
Intracellular machinery responsible for Apoptosis seems to be similar in all animal cells
Depends on a family of Proteases that have a cysteine at their active site and cleave their target
proteins at specific aspartic acids (caspases)
Procaspase —> Caspase (cleavage at Aspartic acids by other cascade)
Once activated, caspases cleave and activate other procaspases, leading to an amplifying proteolytic
cascade
Activated caspases then cleave other key proteins in the cells.
Cleave Nuclear Laminas, causing degeneration of NE
Degrades protein that keeps DNA degrading enzymes in inactive forms — thereby activating
DNA degradation
Degeneration of mitochondria

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Cells infected with viruses are killed by Cytotoxic T Lymphocytes by inducing apoptosis
CTLs induce apoptosis in each other and even in themselves in order to prevent effector cells from
attacking body constituents, when cell mediated immune responses wane. Defects in Apoptotic
machinery leads to auto immune disorders.
Cells with DNA damage: Damage to genome can cause a cell to disrupt proper embryonic
development leading to birth defects and become cancerous. Cells respond to DNA damage by
increasing production of p53 which is an inducer of Apoptosis. Cancer cells often have mutations in
the p53 gene.
Radiation and chemicals can induce apoptosis
HPV produces a protein that inactivates p53 — makes the cell resistant to apoptosis, and hence
cancerous
B cell Leukemia and Lymphomas express high levels of BCL-2 which block Apoptotic signals that
may be received.
NECROSIS
Cells can maintain homeostasis within a narrow range of conditions through physiological and
morphological adaptations
Extreme Stress — Cell cannot adapt — Irreversible injury and death
What factors could lead to cell death?
Oxygen Deprivation: Loss of oxygen carrying capacity can lead to anaemia and CO poisoning
Ischemia: Loss of blood supply from impeded arterial flow and reduced venous drainage. Compromises
supply of oxygen and metabolic substrates.
Physical Injury due to mechanical trauma and temperature extremes
Infections: ranging from virus to large worms
Toxic metals, such as Cyanide block oxidative phosphorylation in mitochondria
Necrosis
Changes that follow cell death in living tissue

Denaturation of intracellular protein
Enzymatic digestion of the cells
Autolysis: Enzymes from Lysosome of same cell
Inflammatory Response: Lysosomes of Leucocytes
Show increased eosinophilia — loss of normal basophilia, increased binding of eosin to denatured
cytoplasmic proteins
Enzymes digest organelles — cytoplasm becomes vacuolated
Dead cells are replaced by large, whorled, phospholipid masses called Myelin figures — phagocytosed
by other cells, degraded into fatty acids

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Necrotic cells
Have discontinuities in Plasma and Organelle membranes

Aggregates of denatured protein
Nuclear changes — non specific DNA breakdown
Pycnosis — Nuclear shrinkage and increased basophilia
Karyorrhexis — Fragmentation of nucleus
Types of Necrosis
Coagulative Necrosis: Preservation of basic outline of cell for some days. Denaturation processes are
dominant
Liquefactive Necrosis: Complete digestion of dead cells. Enzymatic action is dominant leading to a
liquid viscous mass
Caseous Necrosis: Fragmented, coagulated cells and amorphous debris. Enclosed within a distinctive
inflammatory border
Dystrophic Calcification: If Necrotic cells and cellular debris are not cleared promptly, they tend to attract
Calcium salts and minerals and become calcified.
Apoptosis Necrosis
Regulated form of cell death

Leads to un-programmed cell death.
Characterised by chromatin condensation and
Cells swell and rupture
cell shrinkage in the early stage.
The released intracellular contents can
Nucleus and Cytoplasm fragment, forming
damage surrounding cells and cause
membrane bound apoptotic bodies which can
inflammation
be engulfed by phagocyte
Initiated by direct cell damage mostly
Initiated by signal transduction process.
physically
Energy is needed. Active process
Energy is not needed. Passive process.
Does not cause inflammation
Causes inflammation
Cell size is enlarged by swelling
Cell size shrinks
Ends with fragmentation of cell into smaller
Ends with total cell lysis
bodies
PM is disrupted
PM is intact
Only pathologic
Both Physiologic and pathologic
Degrowth
Entire animal becomes smaller when deprived of food, regressing until it resembles a diminutive adult
No adverse physiological outcomes
Planarian degrowth is due to decrease in cell number rather than decrease in cell size — most likely a
decrease in stem cell proliferation, but basal mitotic growth rates are maintained, cell death rate
increases
Whole animal size reduction has been reported for a wide range of invertebrates, principally
coelenterates and flatworms
Reversible when nutrition is available again
AGEING
Basic Concepts of Ageing

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Ageing can be defined as the time related deterioration of the physiological functions necessary for
survival and fertility, which in turn increases the probability of death
Characteristics of ageing affect all individuals in a species
It starts after attainment of sexual maturity in most species
Wild animals rarely show signs of ageing. Random mortality from starvation, predation, infectious
disease or harsh environment kills off most animals before they begin to show signs of ageing.
Cells like neurons, skeletal and cardiac muscle which are no longer in the cell cycle degrade with
ageing. This effect is less visible in cells that are replenished by mitosis throughout life such as blood
and epithelium
Life Expectancy: Defined as the age at which half the population still survives (population specific, instead
of just species specific). People normally begin to age in the 2nd half of their life expectancy span
Ageing in Invertebrates
Sponges and Corals: No sign of ageing, because there is constant replacement of old cells by new ones
Lobsters: never stop growing, and show little decline in fecundity with age
Drosophila: Calorie restriction increases life span (also in mammals), certain single genes can extend
life span
Ageing in Vertebrates
Fishes, Amphibians and Reptiles have long lifespans and continue to grow throughout life
Birds and Mammals have a fixed adult size and show signs of ageing
Causes of Ageing
Accumulation of cellular and genetic damages

Oxidative Damage
Ageing is a by-product of normal metabolism
2-3% of oxygen atoms are reduced insufficiently to form reactive oxygen species (ROS)
— super oxide, hydroxyl radical, hydrogen peroxide.
ROS can oxidise and damage cell membranes, proteins and nucleic acids
Drosophila that have been mutated to overexposes Catalase enzyme which degrades
Peroxides, live 30-40% longer.
Vitamins E and C are ROS inhibitors, addition of Vitamin E to diet of flies and nematodes
increases lifespan
Accumulation of Cross Linked proteins
As a person ages, the normal protein folding machinery in the cytoplasm increasingly
malfunctions.
Results in misfolded and cross linked proteins
Such proteins do not participate in cellular functions and are difficult to degrade as well.
Errors of Translation
Leads to misfolded and non-functional proteins.
Harm the normal cellular physiology in very much the same way as cross linked proteins
Accumulation of Senescent Cell
If telomeres get too short, chromosome abnormalities occur and cells tend to get
senescent.
Telomere shortening is faster in short lived species
When human fibroblasts are placed in a culture medium, they proliferate at first, but
eventually a stage comes when their rate of mitosis slows down and finally stops. The cells
live for a hill but cannot pass from G1 to S phase. This is called Replicative Senescence.
Genetic instability by accumulation of mutations
Point mutations accumulate: faulty and inefficient proteins/enzymes

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Mutations in genes involved in DNA replication, would lead to further increase in

mutations
Species with more efficient DNA repair mechanisms live longer
Mitochondrial Genome Damage
Mutation rate in mitochondria is 10-20 times faster than nuDNA.
Mutations in mitochondria could lead to
Defects in energy production
Production of ROS by faulty electron transport
Apoptosis
Age dependent declines in mitochondrial functions are seen in many species
There are hot spots for age related mutation in mitogenome.
Further, genomes with these mutations have a higher replication frequency, meaning over
time mitochondria with mutated genomes outcompete wild type in the cell.
Genetic Programming
Gerontogenes: genes that affect ageing. e.g. TOR, DAF/FoxO
Klotho gene in mice suppresses ageing phenotypes — mutation leads to early onset of old
age symptoms
Hutchinson Gilford Progeria syndrome causes children to age rapidly and die. It is caused
by a dominant mutant gene.
Increased life span of yeast subjected to calorie restriction requires a gene SIR2 (Silent
Information Regulator) — encodes Sir2 Histone Deacetylase which removes acetyl groups
from histone proteins. Increasing activity of Sir2 extends the lifespan of yeast, C. elegans
and Drosophila by condensing the chromatin which prevents mutations due to strand
slippage.
Drosophila with mutations in Methuselah gene live longer
Drosophila that have been mutated to overexposes Catalase enzyme which degrades
Peroxides, live 30-40% longer.
DAF2 gene codes for cellular receptors for insulin and insulin like growth factor.
Suppressing the gene can extend lifespan by 100% in adult worms.
Reduced activity of growth hormone in rats enhances lifespan
Diet, Social and Lifestyle factors
Tobacco, Alcohol and Narcotics
Obesity and chronic hyper-calorific diets
Chronic stress
Chronic Malnutrition
Sedentary Lifestyle
Exposure to strong radiations
Chronic infections
EYE AND HEART
DEVELOPMENT OF EYE
Development of eye begins during late neural stage soon after differentiation of primary brain vesicles.
Initially an outgrowing or protrusion from the Diencephalon extends, known as optic vesicles.
Later it gets constricted to form an optic stalk, connecting brain and optic vesicles. The optic nerve is
the basis of optic nerve formation
Vesicle extends till it touches the epidermis (surface ectoderm). The outer wall of the vesicle
immediately flattens followed by invagination to form a double walled Optic Cup.
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The place where the optic vesicle touches the surface ectoderm is the place that will gradually develop
into lens plate and will get separated from surface ectoderm in the future.
Activation of head ectoderm’s latent lens forming ability and positioning of lens in relation to the retina
is achieved by optic vesicle.
Optic Vesicle becomes the optic cup after meeting the head ectoderm to differentiate into two layers
Cells of the outer layer produce melanin pigment — form pigmented retina
Cells of the inner layer proliferate rapidly and generate a variety of glia cells, ganglion cells,
inter neutrons and light sensitive photo receptor neutrons, called Neural retina (includes light and
colour sensitive photo receptors — rods and cones)
Lens is entirely ectodermal in origin. Cells on each side of the lens plate increase and invaginate
inwards to form lens pit/vesicle. Lens vesicle is pushed into the optic cup and gets detached from the
surface ectoderm.
Remainder of the surface ectoderm develops into corneal epithelium by bridging the gaps.
Space between surface ectoderm and lens placode is invaded by the mesodermal layer. Iris and Ciliary
body develop from anterior portions Optic Cup and surrounding Mesenchyme.
Iris is infront of the lens — pigmented and muscular tissue. Iris muscles control the size of the pupil
Blood vessels are mesodermal in origin
Small grooves form above and below the eye. As these grooves deepen, eyelid folds develop, above
and below the eye.
DEVELOPMENT OF HEART
One of the first structures to form during Organogenesis.

Stages: Heart tube formation, Looping, Chamber Development

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Heart Tube Formation
As embryo folds in its lateral dimension, it causes the lateral edges of the germ disk to approach each
other until they meet, causing the embryo to acquire a tubular form.
Two outer endocardial tubes will come close to each other in the median of the embryo, ventral to the
primitive gut, and start fusing cranially to caudally — forms a single median tube (primitive heart
tube)
Primitive heart tube is of mesodermal origin. First established as a single tube consisting of two
epithelial layers — inner endocardium (endothelial sheet) and outer myocardium (contractile)
Single tubular heart develops many constrictions outlining future structures.
Cranial most area is the Bulbus Cordis
Extends cranially into Truncus arteriosus. Connected to Aortic sac and through Aortic arches to
Dorsal Aorta
Primitive ventricle is caudal to the Bulbus cordis and Primitive atrium is caudal to primitive
ventricle
During development, it gets divided into atrial and ventricular chambers. 2 chambered heart is the basic
adult form in fish, but in higher vertebrates (birds and mammals), looping and further partitioning gives
rise to the four chambered heart.
Looping
Heart tube continues stretching and by Day 23 Cardiac Looping begins

Atrial position moves left
Chamber Formation
Later development of the heart invovles formation of separate chambers — 4 in case of mammalian
heart
Separation of atrium from ventricle is accomplished when cells from Myocardium produce
transforming growth factors that detaches cells from endocardium and enters cardiac jelly (endocardial
cushion cells) — form the atrioventricular septa
Septa initially have openings for blood to cross, but later they close
Pulmonary artery connects the right ventricle and aorta connects the left ventricle

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PLACENTA
Placental Mammals are mammals whose young are nourished for an extended period of time by an organ in
the womb called a placenta.
Placenta
Placenta is a composite structure of embryonic and maternal tissues that supplies nutrients to the
developing embryo.
It is a flattened circular organ in the uterus of pregnant mammals that nourishes and maintains the
foetus through the umbilical cord
Placenta attaches the foetus to uterine wall and allows the mother to exchange nutrients with the
developing offspring and get rid of waste.
Because of Placenta, Eutherians develop longer in the womb than other mammals, and are more
independent at birth
Parts of placenta
Chorion
Embryonic derived portion of the placenta.
Composed of Trophoblasts, which make the outer cell layer of the Blastocyst.
During implantation, trophoblasts multiply in number and extend into the uterine wall.
Eventually form Chorionic Villi — fingerlike structures of the placenta composed of Embryo
derived trophoblasts
Chorionic villi are surrounded by maternal blood which comes into direct contact with
Embryonic Trophoblast cells.
Intervillous Space
Part of the Placenta that surrounds the Chorionic villi and contains maternal blood
Development of Placenta
After implantation of embryo to the uterine wall, blastocyst of embryo penetrates deep into the uterine
wall.
Extra embryonic membrane Chorion, that surrounds the Trophoblast, is produced into finger like
projections called Villi, that penetrate into depressions in the Uterine wall.
Villi are penetrated by Allantois which forms connective tissue and blood vessels of the foetal
placenta.
Allantochorionic villi become highly vascular and grow into the Uterine wall.
This interdigitate structure of embryonic (allantochorionic villi) and maternal part (uterus wall) is
called the placenta
Trophoblast differentiation
After chorionic villi are established, many of the Trophoblasts fuse together to form very large
synctiotrophoblasts — large, multi nucleated trophoblasts that are formed by fusion of several
smaller cells.
Synctiotrophoblasts are the main component of Chorionic Villi and are the cells that are in direct
contact with maternal blood.
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They are the major source of most placental hormones

Other trophoblasts form columns of cells which serve to anchor the placenta to the uterus. These
junctional Trophoblasts secrete ECM proteins that help them anchor the placenta to the uterus.
Placenta invasion
Trophoblast cells of the placenta in many species have acquired mechanisms to invade the uterus,
inclusive of its blood vessels, to establish efficient fetomaternal exchange of molecules.
Invasion is strictly controlled both spatially and temporally (In humans, usually continues until
midgestation)
Key mechanisms underlying various steps in trophoblast invasion are
Attachment to the basement membrane, most likely by binding to laminin
Detachment from the basement membrane matrix, a process requiring the presence of complex-
type oligosaccharides on the cell surface
Breakdown of basement membrane components, mediated by secretion of metalloproteases
(such as type IV collagenases) and serine proteases (plasminogen activator)
Trophoblast invasiveness in situ is controlled by the microenvironment
Anti-invasive factors by the decidual tissue of the uterus
TIMP (tissue inhibitor of metalloproteases), which neutralizes metalloproteases
TGF-beta (transforming growth factor-beta) - induces TIMP-1 secretion and promotes
differentiation of invasive trophoblast cells into multinucleated giant cells, which are presumably
noninvasive
Thus, TGF-beta provides the key control of trophoblast invasiveness
Based on types of Foetal membranes involved
Yolk Sac Placenta/False Placenta/Chorio-Vitelline Placenta

Formed by Yolk Sac and Chorion.
Primitive placenta formed by few primitive marsupials like Didelphis (Opposum) and
Macropus (Kangaroos, Wallaby)
Connection is established between uterine wall and that part of chorion which is lined with yolk
sac and has a network of vitelline blood vessels of the yolk sac.
Allantois remains relatively small
True Placenta/Chorio-Allantoic Placenta
Connection is established between Uterine wall and chorion lined with Allantois and has a
network of allantoic blood vessels
Yolk Sac remains rudimentary and non-functional.
Found in some marsupials, and all Eutherian mammals
Based on distribution of villi on Chorion
Diffuse: In some mammals, villi remain scattered all over the surface of the chorion and their placenta
are correspondingly extensive
Cotyledonary: Villi are found in groups or patches, while the rest of the Chorion surface remains
smooth. Found in ruminant ungulates (cattle, sheep, deer)
Zonary: Villi are in a belt or girdle like and around the middle of the Blastocyst. Occurs in Carnivores
like cats and dogs.
Discoidal: Villi are restricted to a circular disc or plate on the dorsal surface of blastocyst. Occurs in
insectivores, bats, rodents, rabbit etc.
Metadiscoidal: Special variation of discoidal placenta in which villi are at first scattered but later
restricted to one or few discs. Humans have Metadiscoidal placenta.
Based on degree of association

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Non Deciduate: Contact between foetal and maternal tissue is superficial. At the time of birth, no
injury to the uterine wall and no bleeding occurs. It is found in pigs, cattle, horse etc.
Deciduate: Extensive contact between foetal and maternal tissue. Chorionic villi is highly branched.
During parturition, a large part of uterine tissue is lost. Found in man, rabbit, dog, cat
Based on Histology
Epitheliochorial Placenta: Uterine epithelium of the mother makes contact with Chorion of Embryo.
Chorioallantoic villi lie in the crypts or depressions of the uterine wall. Most primitive type of placenta.
e.g. Marsupial and Ungulates.
Syndesmochorial Placenta: Chorionic Villi erode the uterine wall, so that the uterine epithelium is
ruptured and chorionic epithelium lies in contact with connective tissue of the uterine wall. e.g.
Ruminant Ungulates
Endotheliochorial Placenta: Not only uterine epithelium, but the uterine connective tissue is also
eroded. Chorionic epithelium of villi comes in contact with endothelium of maternal blood vessels. e.g.
Dog, Cat and other carnivores
Haemo Chorial Placenta: Uterine epithelium, connective tissue and endothelial wall of maternal
blood capillaries are eroded and chorionic epithelium comes in contact with maternal blood
Haemo Endothelial: Uterine epithelium, Uterine connective tissue, Maternal blood capillary
endothelium and trophoblastic epithelium are all eroded. Foetal capillaries lie in maternal blood
Functions of Placenta
Blood Flow and Nutrients
Maternal blood flows from the mother’s circulatory system, through the intervillous space and then re-
enters the mother’s blood vessels
Foetal blood flows through the foetus into two main arteries in the umbilical cord, through the
capillaries network of the chorionic villi and are returned to the foetus by the umbilical vein.
Maternal blood that enters the placenta is nutrient and oxygen rich.
In the placenta, foetal blood in the villi is poor in oxygen/nutrients and rich in carbon dioxide and
water, while maternal blood in the intervillous space is rich in nutrient/oxygen and poor in water.
Nutrients and Oxygen diffuse from maternal blood into the foetal blood while wastes diffuse from
foetus blood into maternal blood.
Mother’s lungs, kidneys and liver remove these wastes from her blood
Functional importance
Nutrition: Transports nutrients through blood from mother to the embryo

Respiration: Gaseous exchange between mother and embryo
Excretion: Nitrogenous wastes of embryo diffuse into maternal blood through placenta
Anchorage: Villi attach embryo to the uterine wall
Immunity: Antibodies developed in blood of mother against diseases like small pox, measles etc. are
passed to embryo, which develops passive immunity
Storage: Substances like fat, glycogen, iron etc.
Endocrine Function: Placenta functions as an endocrine gland secreting Oestrogen, Progesterone, FSH,
LH, Relaxin
Protection: Barrier for transport of certain pathogens from mother to foetus. But bacteria and virus can
pass
Nourishment after birth: In mammals like rabbit, mother eats the placenta after birth
Drugs administered to mother can be transported to embryo through placenta.
BIOGENETIC LAW

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Evo-Devo concept refers to study of evolutionary relationships by comparing developmental stages across
different animals.
Ernst Haeckel: “Ontongeny recapitulates Phylogeny”
Proposed that embryonic development of an individual organism followed the same path as
evolutionary history of its species.
Ontogeny is the growth and development of an individual organism
Phylogeny is the evolutionary history of a species
Development of advanced species passes through stages represented by adults of more primitive
species
In this view, creation of new phyla is a step towards the completion of human development. Stages are
added sequentially until a human being has evolved.
This was based on comparative study of embryos of the same age of vertebrates of the same age of
vertebrates. Such comparative studies reveal remarkable similarity and at the same time, displays
progression of complexity.
Laws given by Haeckel
Law of Correspondence: Zygote of an advanced species corresponds to adult of a primitive species.

e.g. Human zygote was represented by 'adult' stage of the protists. Haeckel considered an extinct
organism Gastrea (2 layered sac) corresponding to the Gastrula, which he considered the ancestor of all
metazoan species.
Law of Terminal Addition: New species evolved by adding a step at the end of 'primitive' species.
Thus, humans evolved when embryo of the next highest ape added a new stage. Thus, phylogeny was
linear, instead of being branched. (Different from Darwinian Evolution)
Law of Truncation: Preceding development could be foreshortened. This was needed since for the
model, since not all stages was observed in all animals. It was postulated that this was essential to
prevent gestation time from being enormous.
Analysis
Haeckel’s synthesis tried to combine works of Darwin and Lamarck.

Though there are stages that related embryos do share, embryos do not pass through adult stages of
other animals. e.g. All vertebrate embryos pass through a stage in which there are embryonic gill slits.
Only fish elaborate them into new gills. Other embryos do not pass through a stage where it has
structure of an adult fish
Humans share ancestors with other taxa, but stages of human embryonic development are not
functionally equivalent to the adults of these shared common ancestors.
Development of features leading to speciation is non-linear
Baer’s Laws
Reported by Von Baer in 1828. Based on detailed study of chick development, and comparing it with
embryos of other vertebrates.
1. General features of a large group of animals appear earlier in development that the specialised features
of a smaller group
All developing vertebrates appear very similar shortly after gastrulation
It is only alter that special features corresponding to class, order and species emerge.
All vertebrate embryos have gill arches, notochords, spinal cords and primitive kidneys
2. Less general characters are developed from more general, and finally most specialised characters
appear
All vertebrates initially have the same type of skin.

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Later they differentiate into fish scales, reptile scales, bird feathers or the hair, claws and nails in
mammals.
Early development of the limb is essentially the same in all vertebrates. Only later, do the
differences between legs, wings and arms become apparent
3. Embryo of a given species, instead of 'passing through' adult stages of lower organisms, departs more
and more from them.
Visceral clefts of embryonic birds and mammals do not resemble gill slits of adult fish in detail.
They resemble the visceral clefts of embryonic fish and other embryonic vertebrates. While fish
preserve the clefts and elaborate them into true gill slits, mammals convert them into structures
such as Eustachain tubes (between ear and mouth)
4. Early embryo of a higher animal is like early embryo of lower animal, and not like adult of the lower
animal
Human embryos do not pass through a stage where they are functionally equivalent to adult fish
or reptile.
Rather, they share features with fish embryo and alter diverge in functions, none of them passing
through stages of others.
Three germ layers giving rise to different organs is a constant.
STEM CELLS
Stem cell is a cell capable of extensive proliferation, creating more stem cells as well as more
differentiated cellular progeny.
Potency: Capacity to differentiate into specialised cell types
Self Renewal: Ability to go through numerous cycles of cell division while maintaining the
undifferentiated state.
Development of cells derived from stem cells gives rise to differentiated cell types.
Stem cells are retained in adults fro
Tissue Renewal: Blood cells, intestinal crypt cells, epidermis and spermatocytes have
populations in a steady state equilibrium. Stem cells are used to balance the cell loss
Tissue Regeneration: Produce more stem cells or differentiated cells when body equilibrium is
stressed by injury or environment. e.g. Production of enormous numbers of RBCs when the body
suffers from Anoxia.
Types of Potency
Totipotent: Can differentiate into embryonic and extra embryonic cell types. These cells can create a
complete, viable organism. Such cells are mostly zygotic (or a few divisions of zygote)
Pluripotent: Descendants of totipotent cells and can differentiate into several unrelated types of cells.
e.g. Cells from any of the three germ layers
Multipotent: Differentiate into a number of cells, but only from a closely related family of cells. e.g.
Blood stem cells
Oligopotent: Can differentiate only into a few cells. e.g. Myeloid stem cells
Unipotent: Can produce only one cell type, but have property of self renewal which distinguishes them
from non stem cells e.g. Muscle cells
Occurrence of Stem Cells
Embryo: ESC are derived from inner cell mass of blastocyst. Pluripotent
Foetus: Derived from germ cells of spontaneously aborted foetus. Pluripotent cells are also found in
foetal brain

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Adult Body: Found in small numbers in various tissues in the adult body (e.g. brain and muscle)
Umbilical cord, Cord blood: blood stem cells.
Induced Pluripotent Stem Cells (iPS)
Engineered from somatic cells that were not pluripotent.

Cell with specialised function has been reprogrammed to an unspecialised state similar to that of an
embryonic stem cell.
ESC and Therapeutic Cloning
ESC are pluripotent. They can be cultured indefinitely, yet kept in an undifferentiated state.
Can be obtained from inner cell mass blastomeres of human blastocysts, such as those left over from in
vitro fertilisation
Can be derived from germ cells derived from spontaneously aborted foetus
Applications
Tissue regeneration to deal with degenerative diseases

Potential situations
New neutrons for degenerative brain diseases (like Alzheimer’s or Parkinson) or spinal
injuries
Pancreas for diabetics
Blood cells for Anemia
Heart cells
Replenishments for immune disorders
Mouse ES cells have been manipulated to form insulin secreting cells, muscle stem cells and
glial stem cells. Dopaminergic neutrons from ES cells have been shown to significantly reduce
symptoms of Parkinson disease in rodents.
ES cells are induced to become neural stem cells by growing them on mesoderm and providing
FGF2 in the medium. Neural stem cells can be induced to form ventral neural SC by adding a
paracrine factor Sonic Hedgehog. Further exposure to FGF and BDNF produced cells that had
characteristics of Dopaminergic neutrons
Human ESC derived from germ cells can cure virus induced paraplegia in rats. They differentiate
into new neutrons and produce paracrine factors that prevent death of existing neurons.
Possible limitation is immune rejection of foreign SC
Therapeutic Cloning
Nucleus from patient is inserted into an enucleated oocyte.
Resulting embryos grown in vitro till it has developed an inner cell mass
Cells from inner cell mass are cultured to generate stem cells that are genetically identical to the
patient.
Resolves the issue of immune rejection
Multipotent Adult Stem Cells
Hematopoietic stem Cells: give rise to all kinds of blood cells — RBC, B Lymphocytes, T
Lymphocytes, NK cells, Neutrophils, Basophils, Eosinophils, Monocytes, macrophages and platelets
Bone marrow stroll Cells: Give rise to Bone Cells, Cartilage cells, fat cells and other kinds of
connective tissue such as those in tendons
Neural Stem Cells: Give rise to 3 major cell types — Neurons, astrocytes and oligodendrocytes
Epithelial stem cells in the lining of the digestive tract occur in deep crypts and give rise to several cell
types — absorptive cells, goblet cells, paneth cells, entero-endocrine cells.
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Skin stem cells occur in basal layer of the epidermis and at the base of hair follicles. Epidermal stem
cells give rise to keratinocytes which migrate to the surface of the skin and form a protective layer.
Contribute to tissue specific regeneration (e.g. bone marrow transplantation which uses hematopoietic
SC)
Single blood cell can reconstitute the entire blood system
Limitations compared to ES cells
Not as easy to use as ES cells

Difficult to isolate: Fewer than 1/1000 cells in an organ
Low rate of cell division and do not proliferate readily
Transgenic Stem Cells
Combination of Somatic Cell Nuclear Transfer and Gene Therapy

Collect adult cells from patient (mutant for some gene) and insert into enucleated egg — blastocyst —
ES cells
Converted to wild type by homologous recombination and re-inserted into patient to re-populate the
faulty cell line
Achievements of Stem Cell Therapy
Administration of drugs to increase stem cell division rate in brain, and direct the survival and
differentiation of newly formed cells could be successful in treating stroke and traumatic brain injury.
Parkinson’s disease: foetal transplants of dopamine producing cells
Injecting neural stem cells into brains of drugs can be successful in treating cancerous tumours
Multipotent Adult SC from umbilical cord to treat spinal injuries
Paralysis: ESC injection can regenerate neutrons and myelin sheath
Retinal stem cells into damaged eyes to restore vision (cornea transplant in LVPEI)
Use stem cells in hair follicles to grow skin for grafts
IVF and EMBRYO TRANSFER
IN VITRO FERTILISATION
Eggs and Sperms are retrieved from male and female partners and placed together in a petridish for
fertilisation.
After the zygotes start dividing, they are transferred into the uterus of the female, where implantation
and embryonic development occur like in a normal pregnancy
Success rates are higher than normal pregnancy rates in any given month when woman is under age 40
and there are no sperm problems
Limitations
Rate of delivery of live babies per oocyte retrieval depends on age of female partner.
After 40y of age, success rate is less than 5%. Most likely declines due to declining quality of eggs.
Transfer of more embryos increases the risk of multiple births. Multiple birth infants are pre disposed
to many health problems like malformations, infant death, premature delivery etc.

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Method
Ovary stimulation
Gonadotrphins or Anti-estrogens are used to hyper stimulate the ovaries to produce several
mature oocytes.
Having several mature eggs for IVF increases the possibility that atleast one would result in
pregnancy
Collection of eggs
When the follicle has matured (but not yet ruptured), the physician attempts to retrieve as many
eggs as possible (secondary oocytes).
Aspiration pipette is guided to each mature follicle and the oocyte is sucked out.
Mature and healthy oocytes are transferred to a sterile container to await fertilisation
Sperm Collection
Sperm sample is collected from the male partner 2h before the female partner’s oocytes are
retrieved.
Sperm washing procedure is used to capacitate the sperm and select the healthiest and most
active sperms.
Fertilisation
Sperm are placed in a petridish with oocytes and the gametes are incubated at room temperature.
If fertilisation is successful, eggs will be ready to divide and the resulting embryos would be
ready to be transferred into the uterus
Embryo Transfer
Embryos are placed in a catheter.
The catheter is inserted through the vagina to place them directly in the uterus.
If implantation to uterus fails, physician may lyse a small hole into the bona pellucid prior to
inserting the embryo into the uterus.
Variations in IVF
Intra Cytoplasmic Sperm Injection

Single sperm is injected into the cytoplasm of the egg.
Used when the male partner has extremely low numbers of normal viable sperm. (Due to genetic
factors, blockages in ducts, injuries to reproductive organs
Gamete Intra Fallopian transfer
Sperm are injected into the oviduct at a time when oocytes are ovulated.
Generally recommended when the sperm cannot reach the oocyte in the oviduct.
Occurs naturally within the female partner’s body
Zygote intra fallopian transfer
Fertilisation takes place outside the body in a petridish.
Resulting zygotes are transferred into the female partner’s Fallopian tube.
EMBRYO TRANSFER
Embryo transfer is the step in IVF, where one or several embryos are placed into the uterus of the
female with the intent to establish a pregnancy.
Embryos can be fresh from fertilised egg cells or frozen
Woman with eggs but no uterus — requires a surrogate womb
Woman with no eggs but uterus — requires an egg donor
Cryopreservation of Embryos
Freezing and storage of embryos in liquid nitrogen (-196ºC)

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Embryos are first treated with a suitable concentration of glycerol (Cryoprotectants) which protects
them from injury during freezing and thawing.
They are then cooled at a slow rate to -38ºC, employing a programmable controlled rate freezer.
Then the embryos are plunged into liquid nitrogen and stored at -196ºC.
The embryos are thawed at a very rapid rate by immersing the ampoule carrying them in a water bath
maintained at 0ºC.
Uterine Preparation
Uterine lining (endometrium) needs to be appropriately prepared so that the embryo can implant.
In a natural cycle, the embryo transfer takes place in the luteal phase, when the lining is appropriately
developed to receive the embryo.
Recipient is given oestrogen preparations (2 weeks) and then a combination of oestrogen and
progesterone, so that the lining becomes receptive for the embryo.
Time of receptivity is called the 'implantation window'
Timing of Transfer
Embryos are generally transferred to the woman’s uterus at the 2-8 cell stage (48-72h after
fertilisation).
Embryos that reach the day 3 cell stage are tested for chromosomal or specific genetic defects prior to
transfer
Procedure
Speculum in the vagina to visualise the cervix

Cervix is cleaned with saline solution or culture media.
A transfer catheter is loaded with the embryos. It is inserted through the cervical canal and advanced
into the uterine cavity where the embryos are deposited.
Abdominal ultrasound is used to ensure correct placement.
Elective Single Embryo Transfer (e-SET) lowers the risk of multiple pregnancies
Post Transfer
Injection of HCG to help the ovaries produce more progesterone during the embryonic implantation
phase.
Outcome of IVF cycle depends on quality of embryos and post transfer hormonal support
Embryo Transfer in Cattle
Chief objective is to obtain several progeny per year from a single female of superior genotype.
In India, most cattle are of inferior genotype with low productivity. Superior genotypes are limited in
number and expensive.
Artificial Insemination involves 50% genes from superior genotype male but 50% from inferior
female.
In ETT, inferior females are only used as surrogate mothers, thus allowing embryo to be 100% superior
genotype.
Procedure
Genetically superior high productivity female and male serve as source of embryos to be transferred.
Healthy, young females are selected to be recipients of embryos to be transferred.
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Donor females are treated with appropriate doses of gonadotrophin — FSH, LH — to increase the
number of ova released at the time of ovulation (Super Ovulation). Under optimum treatment
conditions, single female can provide upto 15 embryos in a single oestrus cycle.
Donor female is artificially inseminated using semen from a genetically superior male
Fertilised eggs/young embryos are collected by flushing the uterus os superior donor females with a
special nutrient solution (7 days after insemination). Embryos are examined under a stereoscopic
microscope and normal looking, healthy embryos are selected.
Selected embryos are incubated in a special nutrient medium at 37ºC till their transfer into the
surrogate mothers. Or they may be frozen and stored in liquid nitrogen for further use.
Single embryo is transferred into the uterus of each surrogate mother.
Applications of ETT
Rapid rate of multiplication animals of the superior genotype. 36 embryos using ETT in a year
compared to just one by natural methods
Rate of multiplication ca be further increased by splitting embryos which develop into a separate
progeny
Young embryos can be cryopreserved for upto 10 years. Easier to transport and present no quarantine
problems.
Superior cows that cannot bear embryos, can act as donors
Limitations
High degree of expertise is required

Cost of producing each progeny is much higher

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ﬁle:///Users/sashibhusanmishra/Documents/Dev Bio/2B Dev Bio.html 1/63

Starts with division of spermatogonia that line the

During spermatogonial divisions, cytokinesis is not

Secondary spermatocytes undergo Meiosis II and give rise

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Stage of differentiation of the sperm cell, during which it is

Mice lacking BMP8 do not initiate spermatogenesis at puberty

Approximately 200 to 500 million Spermatozoans are released per

Amino acids, Citrates, Enzymes, Flavins, Proteins, Vitamin C

Prostatic ﬂuid gives semen a milky appearance.

When spermatozoa remain in the ﬂuid of the male genital ducts

In-Vitro Capacitation for IVF

External ﬁbre-vascular coat

Why is Oogenesis different from Spermatogenesis?

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Two principal events go on side by side

Unique in its placement of the metaphase plate.

Occurs in the Diplotene stage of Meiotic prophase

Two patterns of ovulation

Ovulation is stimulated by act of copulation

Menstrual cycle in humans is an integration of

Ovarian cycle — mature and release an oocyte

Oogenesis and Ovulation in humans

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Egg and sperm come together outside of the parent’s bodies.

Takes place inside the female body

Sequence of interaction of sperm and egg

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Sperm are attracted to the ﬂuid surrounding the egg.

Zona pellucida in mammals plays a role analogous to vitelline envelope in

During acrosomal reaction, anterior portion of the sperm PM is shed (where

Fertilin proteins in sperm plasma membrane are essential for membrane

Fast Block to Polyspermy Slow Block to Polyspermy

Peroxidase enzyme hardens the fertilisation

Isolecithal: Distribution of yolk is even throughout the egg

Major events during gastrulation

Rearrangement of cells due to morphogenetic movements of the cells of blastoderm

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Fate of primary germ layers

Endoderm: Digestive and respiratory tracts and associated structures

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Hypoblast is separated by underlying yolk by a subgerminal cavity.

Deﬁnes the axes of the embryo. Extends from P to A.

Morphogenetic and Gastrula Movements

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Vital Dye Marking

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Creating mosaic embryos having different genetic constituents (chimeric embryos)

Methods to Study Cell Lineage

TYPES OF TISSUE GROWTH

Changing of one cell type to another, usually as an adaptation to external environment.

"New Growth" - unregulated cell proliferation as a result of genetic changes.

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Crabs break off their leg on approaching of the enemy

Regenerative capacity in Animal Groups

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Types of Regeneration based on Cellular Mechanism