What are the different types of regeneration? Add a note on regeneration of salamander's limb.

Regeneration is the reactivation of developmental mechanisms in postembryonic life to restore

missing or damaged tissues. Regeneration was first reported in Hydra by Tremble in 1740.
There are several types of regeneration observed in nature:

1. Epimorphosis: This type of regeneration involves the proliferation of undifferentiated cells at

the site of injury, forming a blastema. The blastema then differentiates into the specific tissues
needed to replace the lost structure. Epimorphosis is common in organisms like amphibians
and some invertebrates.
2. Compensatory: Compensatory regeneration occurs when the remaining tissues of an organ
compensate for the lost or damaged portion by growing larger or increasing in activity. Here,
the differentiated cells divide but maintain their differentiated functions. The new cells do not
come from stem cells, nor do they come from the dedifferentiation of adult cells. Each cell
produces cells similar to itself; no mass of undifferentiated tissue forms. This type of
regeneration is often observed in mammals, such as the liver's ability to regenerate after
partial removal.
3. Morphallaxis: Morphallaxis involves the remodelling of existing tissues to regenerate the lost
structure. Instead of forming a blastema, the existing tissues reorganise and differentiate to
replace the lost part. Morphallaxis is common in organisms like Hydra and planarians.

Salamander exhibits epimorphic regeneration which involves dedifferentiation of adult structures in

order to form an undifferentiated mass of cells.

The stages of limb regeneration in the salamander are as follows:

1. Blood and immune cells flood the amputated area, and a blood clot quickly forms.
2. Wounding triggers an activation of stem/progenitor cell proliferation.

3. Epidermal cells along the edge of the cut migrate over the wound to form the wound epidermis.

4. Through cell proliferation and continued migration, the wound epidermis thickens into the apical
epidermal cap (AEC).

5. Signals from the AEC to the amassing population of progenitor cells underneath it foster
development of the regeneration blastema.

6. Continued proliferation and progressive differentiation of the blastema fuel the outgrowth of the
limb regenerate

Include signals involved in the initiation of limb regeneration

What are the different types of metamorphosis in insects ? Add a note on the role of hormones
in insect metamorphosis.
Metamorphosis is a biological process observed in many organisms, particularly in insects and
amphibians, where a profound and often rapid change in body form and structure occurs during
development. It involves a series of distinct stages, each with its own characteristic morphology and
physiological adaptations.
In insects, metamorphosis typically involves a transition from larval stages to adult stages, often via
intermediate stages such as pupae.
There are three major patterns of insect metamorphosis:
 1. Ametabolous
2. Gradual, or hemimetabolous
3. Holometabolous
1. A few insects, such as springtails, have no larval stage and undergo direct, or ametabolous,
development.
Immediately after hatching, ametabolous insects have a pronymph stage bearing the structures that
enabled it to get out of the egg.
The insect looks like a small adult; it grows larger after each molt with a new cuticle, but is
unchanged in form.
2. Grasshoppers and bugs undergo a gradual, or hemimetabolous, metamorphosis. After spending a
very brief period of time as a pronymph (whose cuticle is often shed as the insect hatches), the insect
looks like an immature adult and is called a nymph. The rudiments of the wings, genital organs, and
other adult structures are present and become progressively more mature with each molt.
At the final molt, the emerging insect is a winged and sexually mature adult, or imago.
3. In the holometabolous development of insects such as flies, beetles, moths, and butterflies, there is
no pronymph stage.
The juvenile form that hatches from the egg is called a larva.
The larva (a caterpillar, grub, or maggot) undergoes a series of molts as it becomes larger. The stages
between these larval molts are called instars.
The number of larval molts before becoming an adult is characteristic of a species,
although environmental factors can increase or decrease the number.
The larval instars grow in a stepwise fashion, each instar being larger than the previous one.
Finally, there is a dramatic and sudden transformation between the larval and adult stages: after the
final instar, the larva undergoes a metamorphic molt to become a pupa. The pupa does not feed, and
its energy must come from those foods it ingested as a larva.
During pupation, adult structures form and replace the larval structures. Eventually, an imaginal molt
forms the adult (imago) cuticle beneath the pupal cuticle, and then the adult later emerges from the
pupal case at adult eclosion.
While the larva is said to hatch from an egg, the imago is said to eclose from the pupa.
Role of hormones in insect metamorphosis:
The metamorphosis of insects is regulated by systemic hormonal signals, which are controlled by
neurohormones from the brain (prothoracicotropic hormone (PTTH) in
response to neural, hormonal, or environmental signals).
Insect molting and metamorphosis are controlled by two effector hormones:
1. steroid 20-hydroxyecdysone (20E)
2. lipid juvenile hormone (JH)
1. Steroid 20-hydroxyecdysone initiates and coordinates each molt (whether larva-to-larva,
larva-to-pupa, or pupa-to-adult) and regulates the changes in gene expression that occur during
metamorphosis.
PTTH stimulates the production of ecdysone by the prothoracic gland by activating the RTK
(receptor tyrosine kinase) pathway.
Ecdysone is then modified in peripheral tissues to become the active molting hormone 20E.
Each molt is initiated by one or more pulses of 20E. For a larval molt, the first pulse produces a small
rise in the 20E concentration in the larval hemolymph (blood) and elicits a change in cellular
commitment in the epidermis.
A second, larger pulse of 20E initiates the differentiation events associated with molting. These
pulses of 20E commit and stimulate the epidermal cells to synthesize enzymes that digest the old
cuticle and synthesize a new one.
High levels of JH prevent the ecdysone-induced changes in gene expression
that are necessary for metamorphosis.
2. Juvenile hormone is secreted by the corpora allata. Larval-to-larval molts are produced when
there are large circulating titers of JH.
The secretory cells of the corpora allata are active during larval molts but inactive during the
metamorphic molt and the imaginal molt.
In the last larval instar, the level of JH drops through two mechanisms: the medial nerve from the
brain to the corpora allata inhibits these glands from producing JH, and there is a simultaneous
increase in the body’s ability to degrade existing JH that molt is another larval instar, not a pupa or an
adult.
When the concentration of JH becomes low enough, the 20E-induced molt produces a pupa instead of
a larva. When 20E acts in the absence of JH, the imaginal discs
differentiate and the molt gives rise to an adult.
What is ART ? Explain the process of invitro fertilization.
ART stands for Assisted Reproductive Technology. It refers to medical procedures used to assist
individuals or couples in achieving pregnancy when conventional methods are not successful. ART
encompasses a variety of techniques, including:

1. In vitro fertilization (IVF): This involves fertilizing an egg with sperm outside the body, in a
laboratory dish. The fertilized embryo is then transferred to the uterus for implantation.
2. Intracytoplasmic sperm injection (ICSI): In ICSI, a single sperm is injected directly into an
egg to facilitate fertilization. This technique is particularly useful in cases of male infertility.
3. Gamete intrafallopian transfer (GIFT): GIFT involves transferring eggs and sperm into the
fallopian tubes, where fertilization occurs naturally inside the body.
4. Zygote intrafallopian transfer (ZIFT): Similar to IVF, ZIFT involves fertilizing eggs in the
laboratory, but instead of transferring embryos to the uterus, they are placed directly into the
fallopian tubes.
5. Donor eggs or sperm: ART may involve the use of donated eggs or sperm from individuals
who are not the intended parents.

In vitro fertilisation (IVF) is a process of fertilisation where an egg is combined with sperm outside
the body, in vitro ("in glass"). The process involves monitoring and stimulating a woman's ovulatory
process, removing an ovum or ova (egg or eggs) from the woman's ovaries and letting sperm fertilise
them in a liquid in a laboratory. After the fertilised egg (zygote) undergoes embryo culture for 2–6
days, it is implanted in the same or another woman's uterus, with the intention of establishing a
successful pregnancy.

Steps involved in IVF:

IVF involves several steps – ovarian stimulation/induction, egg retrieval, sperm retrieval, fertilization
and embryo transfer. One cycle of IVF can take about two to three weeks.

1. Ovulation induction:
If a woman using her own eggs during IVF, at the start of a cycle she’ll begin treatment with
synthetic hormones to stimulate her ovaries to produce multiple eggs – rather than the single
egg that normally develops each month. Multiple eggs are needed because some eggs won’t
fertilize or develop normally after fertilization.
In this aspect, some medications are –

For ovarian stimulation - An injection containing FSH, LH or a combination of both. They stimulate
more than one egg to develop at a time.

For oocyte maturation - When the follicles are ready for egg retrieval (generally after 9-14 days), she
has to take HCG to help the eggs mature.

To prepare the lining of the uterus - On the day of egg retrieval or at the time of embryo transfer, the
doctor might recommend that she begin taking progesterone supplements to make the lining of uterus
more receptive for implantation.
Typically, she’ll need 1-2 weeks of ovarian stimulation before the eggs are ready for retrieval. To
determine when the eggs are ready for collection, doctor will likely perform

i. vaginal ultrasound (to see the development of follicles),

ii. ii) blood tests (to measure the response to ovarian stimulation medications – oestrogen levels
increase as follicles develop, progesterone levels remain low until after ovulation).

2. Egg retrieval:
It can be done 34-36 hours after the final injection and before ovulation.

● During egg retrieval, she’ll be sedated and given pain medication.

● Trans-vaginal ultrasound aspiration is the usual retrieval method. An ultrasound probe is
inserted into the vagina to identify follicles. Then a thin needle is inserted through the vagina
and into the follicles to retrieve the eggs.
● The eggs are removed from the follicles through a needle connected to a suction device.
Multiple eggs can be removed in about 20 minutes.

Mature eggs are placed in a nutritive liquid (culture medium) and incubated. Eggs that appear healthy
and mature, will be mixed with sperm to attempt for fertilization. However, not all eggs may be
successfully fertilized.

3. Sperm retrieval:
If the woman using her partner’s sperm, he’ll provide a semen sample at the doctor’s clinic in the
morning of egg retrieval. Other methods such as testicular aspiration (use of a needle to extract sperm
directly from the testicle) are sometimes required. Donor sperm can also be used. Sperms are
separated from the semen fluid in the lab.

4. Fertilization:
Fertilization can be attempted using two common methods:

● Conventional insemination – during this process, healthy sperm and mature eggs are
mixed and incubated overnight.
● Intra-cytoplasmic sperm injection (ICSI) – in this process, a single healthy sperm is
injected directly into each mature egg. It is often used when semen quality or number is a
problem.

5. Embryo transfer:

Embryo transfer is done 2-5 days after egg retrieval:

● The woman might be given a mild sedative.

● The doctor will insert a long, thin, flexible tube (catheter) into patient’s vagina through the
cervix and into the uterus.
 ● A syringe containing one or more embryos suspended in a small amount of fluid, is attached
to the end of the catheter.
● Using the syringe, the doctor places the embryo(s) into the uterus.
● If successful, an embryo will implant in the lining of the uterus about 6-10 days after egg
retrieval.

State various theories of ageing. Give an example of premature ageing syndrome with its
symptoms.
Ageing is a multi-factorial process. Most hypotheses on the underlying mechanisms of the ageing
process involve deterioration of the maintenance of homeostatic metabolic, inflammatory, and/or
redox processes in cells and tissues. There are more than three hundred theories of ageing. But , in
general terms, all can be classified into three big groups: the genetic mutation theories, the wear and
tear theories, and the cellular waste accumulation theories.

General wear-and-tear theory

Dr. August Weismann, a German biologist, first introduced this theory in 1882. He believed that the
body and its cells were damaged by overuse and abuse. The organs and skin are worn down by toxins
in the diet and in the environment. Wear and the tear are not confined to the organs; it also takes place
at the cellular level.

As one gets older, small traumas to the body build up. Point mutations increase in number, and the
efficiencies of the enzymes encoded by our genes decrease. Moreover, if a mutation occurred in a part
of the protein synthetic apparatus, the cell would make a large percentage of faulty proteins.
If mutations arose in the DNA-synthesizing enzymes, the rate of mutations would be expected to
increase markedly. Faulty DNA polymerases in senescent cells have been evident. Likewise, DNA
repair may be important in preventing senescence, and species whose members' cells have more
efficient DNA repair enzymes live longer .

Moreover, genetic defects in DNA repair enzymes can produce premature aging syndromes in
humans.

The Neuroendocrine Theory

This theory was described in the year 1954 by Dr. Vladimir Dilman, a Russian scientist. The
neuroendocrine theory of aging states that ‘‘The effectiveness of the body’s homeostatic adjustments
declines with aging—leading to the failure of adaptive mechanisms, ageing and death.’’

This theory has also been referred to as the aging clock theory. Consistent with this theory, the
hypothalamic pituitary adrenal (HPA) axis, the main regulatory system controlling homeostasis in
humans, loses efficiency with aging. The HPA system works by the interplay of various hormonal
signals that initiate reactions in target tissues coupled with a negative feedback mechanism (the
produced substances inhibit their own production) to allow for fine control of body functions, such as
blood pressure, fluid and electrolyte levels, and body temperature.

This theory proposes that aging is due to changes in neural and endocrine functions that are crucial
for
1. Coordinating communication and responsiveness of all body systems with the external
environment

2. Programming physiological responses to environmental stimuli; and

3. Maintaining an optimal functional state for reproduction and survival while responding to
environmental demands.

Hormones are vital for repairing and regulating the bodily functions, and when aging causes a drop in
the hormone production, it causes a decline in body's ability to repair and regulate itself as well.

Thus, hormone replacement therapy, a frequent component of any anti-aging treatment, helps to reset
the body's hormonal clock and so can reverse or delay the effects of aging and thus keeping young.

The Genetic Control Theory

This theory states that humans are born with a unique genetic code, a predetermined tendency to
certain types of physical and mental functioning, and that the genetic inheritance has a great deal to
say about how quickly one becomes aged and how long lives.
Each person has a biological clock. When that clock goes off, it signals the bodies first to age and
then to die. However, the timing on this genetic clock is subject to enormous variation, depending on
what happens with the growth and on how one actually lives (quality of life, feeding, sanitation and
health care practices).
This theory should be restricted to cases where there is a specific control of the onset of ageing by an
identifiable metabolic process and a functional role for senescence can be demonstrated for that
species, e.g. the rapid ageing and death of the Pacific salmon after spawning.

Waste (undegradable by-products of metabolism) Accumulation Theory of Ageing

In the course of life span the cells produce more waste than they can properly eliminate. This waste
can include various toxins (free radicals, histones, aldehydes, lipofuscins) which when accumulated
to a certain level, can interfere with normal cell function, ultimately killing the cell.

The only mechanism by which these agents are diluted in the cells is cell division. This only applies
to replicative cells. The challenge for multicellular organisms such as the human organism is that
many cell types lose replicative capacity or divide slowly, even though they remain active throughout
the lifespan. These cells, including cardiomyocytes and brain neurons, accumulate metabolic waste
that eventually affects normal cell functioning.

A common by-product of cellular metabolism seems to be lipofuscin (LF): Lipofuscin (age pigment)
is a brown-yellow, electron-dense, autofluorescent material that accumulates progressively over time.
LF is composed of highly oxidized cross-linked macromolecules (proteins, lipids, and sugars) with
multiple metabolic origins. LF cannot be digested in the ubiquitin-proteasome system, because of
highly oxidized proteins, polymeric and highly cross-linked nature. LF cannot be cleared by
exocytosis, and accumulate within the lysosomes and cell cytoplasm of long-lived post-mitotic and
senescent animal cells. LF inclusions from different animal and human tissues (myocardium, brain,
liver, thyroid) are composed mainly of proteins and lipids, 30-70 and 20-50%, respectively. A small
amount of carbohydrates (4.7%) and traces of metals are also found in LF granules.

The accumulation of non-degradable material can occur in the intra- and extracellular environments.

Among the extracellular deposits found in humans, cholesterol-containing plaques and their oxidized
derivatives in blood vessels, as well as protein polymers, such as β-amyloid in the central nervous
system.

An example of premature ageing syndrome is Hutchinson-Gilford Progeria Syndrome (HGPS),

commonly known as Progeria. It is a rare genetic disorder characterized by dramatic, rapid aging
beginning in childhood.

Symptoms of Hutchinson-Gilford Progeria Syndrome (HGPS)

1. Growth Delay: Children with HGPS typically have growth delays evident by around 1 year of
age. They are significantly shorter and weigh less than their peers.
2. Distinctive Facial Features:
● Small Face and Jaw: A disproportionately small face compared to the head.
● Beaked Nose: A thin, pointed nose.
● Thin Lips and Small Chin: Lips may be thin, and the chin often recedes.
3. Skin Changes:
● Aged Appearance: The skin appears thin, dry, and wrinkled.
 ● Loss of Subcutaneous Fat: There is a noticeable lack of subcutaneous fat, leading to a
more skeletal appearance.
● Scleroderma-like Skin: Hardened, tight skin, particularly over the abdomen, thighs,
and buttocks.
4. Musculoskeletal Issues:
● Joint Stiffness and Hip Dislocation: Limited range of motion and joint abnormalities.
● Osteoporosis: Reduced bone density, increasing fracture risk.
● Delayed Dentition: Late appearance and development of teeth.

Short note on Teratogens

● Exogenous agents that cause birth defects are teratogens. They can prevent particular genes or
proteins from functioning or that can activate processes at the wrong places and times.
● Human development is usually divided into an embryonic period (to the end of week 8) and a
fetal period (the remaining time in utero)
● Most organ systems form during the embryonic period; the fetal period is generally one of
growth and modeling
● Maximum fetal susceptibility to teratogens is between weeks 3 and 8
● The nervous system, however, is constantly forming and remains susceptible throughout
development. Prior to week 3, exposure does not usually produce congenital anomalies
because a teratogen encountered at this time either damages most or all of the cells of an
embryo, resulting in its death, or kills only a few cells, allowing the embryo to fully recover.
● The largest class of teratogens includes drugs and chemicals. Viruses, radiation, high body
temperature, and metabolic conditions in the mother can also act as teratogens.
Distinguish between epimorphosis and morphallaxis.

Epimorphosis Morphallaxis

Epimorphosis involves the extensive formation In morphallaxis, regeneration occurs primarily

of new tissues. When a part of an organism is through the reorganization and remodeling of
lost or damaged, a mass of undifferentiated existing tissues rather than the extensive
cells, known as a blastema, forms at the wound formation of new cells.
site. These cells then proliferate and
differentiate to regenerate the lost part.

This process requires significant cell division This process involves little to no cell
and proliferation. The cells in the blastema often proliferation. Instead, cells in the remaining part
derive from dedifferentiated cells at the wound of the organism reorganize to replace the lost
site that have reverted to a more primitive state. structures. Cells often transdifferentiate,
 changing from one type to another to fulfill the
needs of the regeneration.

The regenerated part is usually of the same size The regenerated structure is often smaller than
and structure as the original, indicating a high the original and proportional to the remaining
degree of regulatory control over the part of the organism. Over time, growth and cell
regenerative process. proliferation may restore the organism to its
original size.

Example: regeneration of limbs in salamanders Example: Hydra

Expand: ART- Assisted Reproductive Technology

What are the properties of stem cells? Enumerate different types of stem cells and their
application in regenerative medicine.
Stem cells are undifferentiated cells that have the unique ability to develop into various specialized
cell types and to self-renew, meaning they can divide and produce more stem cells. These properties
make stem cells fundamental to growth, development, and tissue repair in living organisms.
Stem cells are unique cells with remarkable properties that distinguish them from other cell types.
Their key properties include:

1. Self-Renewal
Stem cells have the ability to divide and produce copies of themselves over extended periods without
differentiating. This property is essential for maintaining the stem cell pool throughout an organism's
life.

2. Potency
Stem cells have varying degrees of potency, which refers to their potential to differentiate into
different cell types:
● Totipotent: These stem cells can differentiate into any cell type, including both embryonic and
extra-embryonic tissues (e.g., zygote).
● Pluripotent: Pluripotent stem cells can differentiate into any cell type within the three germ
layers (ectoderm, mesoderm, endoderm) but not extra-embryonic tissues (e.g., embryonic
stem cells).
● Multipotent: These stem cells can differentiate into a limited range of cell types related to
their tissue of origin (e.g., hematopoietic stem cells can become various blood cells).
● Oligopotent: These stem cells can differentiate into a few cell types (e.g., myeloid stem cells
can differentiate into different types of blood cells, like neutrophils and macrophages).
● Unipotent: These stem cells can produce only one cell type but have the property of
self-renewal (e.g., muscle stem cells).
3. Differentiation
Stem cells have the ability to differentiate into specialized cell types. This process is regulated by
internal genetic factors and external signals such as chemicals from other cells, physical contact with
neighboring cells, and certain molecules in the microenvironment.

4. Plasticity
Stem cells exhibit plasticity, meaning they can adapt and change their developmental fate. For
example, under specific conditions, some multipotent stem cells can transdifferentiate, or convert into
a different type of multipotent stem cell.

Types of Stem Cells

1. Embryonic Stem Cells (ESCs)

● Source: Derived from the inner cell mass of the blastocyst (early-stage embryo).
● Potency: Pluripotent, capable of differentiating into any cell type of the body.
● Applications:
● Neurodegenerative Diseases: Potential to generate neurons for treating conditions like
Parkinson's disease and spinal cord injuries.
● Cardiac Repair: Developing cardiac muscle cells for repairing damaged heart tissue
post-myocardial infarction.
● Diabetes: Creating insulin-producing beta cells for diabetes treatment.

2. Adult Stem Cells (ASCs)

● Source: Found in various tissues such as bone marrow, blood, brain, liver, and skin.
● Potency: Typically multipotent, involved in tissue maintenance and repair.
a. Hematopoietic Stem Cells (HSCs)
● Source: Bone marrow, peripheral blood, and umbilical cord blood.
● Applications:
● Blood Disorders: Treatment of leukemia, lymphoma, and other blood-related
conditions through bone marrow transplants.
● Immune System Reconstitution: Post-chemotherapy or in cases of severe
immunodeficiency.
● b. Mesenchymal Stem Cells (MSCs)
● Source: Bone marrow, adipose tissue, umbilical cord tissue etc.
● Applications:
● Orthopedic Repair: Regeneration of bone, cartilage, and muscle tissues for
treating fractures, osteoarthritis, and muscular dystrophy.
● Cardiovascular Repair: Repairing damaged heart tissue and improving
outcomes after myocardial infarction.
● Wound Healing: Enhancing the healing of chronic wounds and skin burns.
● c. Neural Stem Cells (NSCs)
● Source: Brain and spinal cord.
● Applications:
 ● Neurological Diseases: Potential treatments for conditions like Parkinson's
disease, Alzheimer's disease, multiple sclerosis, and spinal cord injuries by
regenerating neural tissue.

3. Induced Pluripotent Stem Cells (iPSCs)

● Source: Somatic (adult) cells reprogrammed to an embryonic stem cell-like state.
● Potency: Pluripotent, similar to ESCs, capable of differentiating into any cell type.
● Applications:
● Personalized Medicine: Creating patient-specific cells for personalized treatment
plans, reducing the risk of immune rejection.
● Disease Modeling: Studying disease mechanisms and developing drugs using cells
derived from patients with specific genetic conditions.
● Tissue Engineering: Generating various tissue types for regenerative therapies,
including skin grafts, retinal repair, and cardiac tissue.

4. Perinatal Stem Cells

● Source: Amniotic fluid, placenta, and umbilical cord blood.
● Potency: Varies but generally considered multipotent or pluripotent.
● Applications:
● Regenerative Therapies: Potential use in repairing or replacing damaged tissues, such
as bone, cartilage, and muscle.
● Immune Modulation: Reducing inflammation and promoting healing in conditions like
autoimmune diseases and organ transplants.

5. Cancer Stem Cells (CSCs)

● Source: Found within tumors and believed to drive cancer growth and recurrence.
● Applications:
● Cancer Treatment: Targeting CSCs to develop therapies aimed at eradicating cancer at
its root, potentially preventing relapse and metastasis.

Regeneration in Hydra.

Hydra Regeneration is Accomplished with Three Different Stem Cells

1. The body wall contains two epithelial cell layers, ectodermal and endo-dermal epithelial cells.
Interstitial stem cells exist within the ectodermal epithelial cell layer.
2. The ectodermal and endodermal epithelial cells proliferate continuously to maintain these
tissue layers, producing differentiated epithelial cells, and are therefore considered to be
distinct stem cells.
3. A third stem cell type, the multipotent, interstitial stem cell can self-renew and produce
neurons, nematocytes, secretory cells, and gametes.
The presence of these stem cells throughout the Hydra's body ensures a continual supply of new cells
for regeneration.

Hydra can regenerate any part of its body, including the head, tentacles, and foot. When the head is
severed, a new head forms at the wound site, and similarly, when the foot is removed, a new foot
regenerates. The head contains a group of cells called the hypostome, which has organizer properties.
This hypostome region can induce the formation of a new head if transplanted to another part of the
body.
The regeneration process restores the head-foot axis, ensuring that new structures form in the correct
positions. Every portion of the hydra’s body column along the apical-basal axis is potentially able to
form both a head and a foot. The animal’s polarity, however, is coordinated by a series of
morphogenetic gradients that permit the head to form only at one place and the basal disc to form
only at another.
When hypostome tissue from one hydra is transplanted into the middle of another hydra, the
transplanted tissue forms a new apical basal axis, with the hypostome extending outward. When a
basal disc is grafted to the middle of a host hydra, a new axis also forms, but with the opposite
polarity, extending a basal disc. When tissues from both ends are transplanted simultaneously into the
middle of a host, either no new axis is formed or the new axis has little polarity.
These experiments have been interpreted to indicate the existence of a head activation gradient
(highest at the hypostome) and a foot activation gradient (highest at the basal disc).

Cellular and structural changes during head regeneration in Hydra

Extracellular matrix (ECM) is retracted immediately upon decapitation followed by change in
endodermal cell shape. This is succeeded by wound closure and secretion of ECM components
completing the early stages of head regeneration.

The major head inducer of the hypostome organizer is a set of Wnt proteins acting through the
canonical β-catenin pathway. These Wnt proteins are seen in the apical end of the early bud, defining
the hypostome region as the bud elongates. If the Wnt signaling inhibitor GSK3 is itself inhibited
throughout the body axis, ectopic tentacles form at all levels, and each piece of the trunk has the
ability to stimulate the outgrowth of new buds. When the hypostome is brought into contact with the
trunk of an adult hydra, it induces expression of the Brachyury gene in a Wnt-dependent manner.

What is ART? Write a note on embryonic stem cells.

ART stands for Assisted Reproductive Technology and consists of medical techniques used to aid
individuals in achieving pregnancy. These technologies address infertility issues.
Common Methods include:
1. In vitro fertilization (IVF) – one of the most common forms of ART is IVF. Here fertilization
occurs outside the body.
2. Gamete intrafallopian transfer (GIFT) – It involves the transfer of sperms and eggs into the
fallopian tube of a woman where fertilization takes place.

3. Zygote intrafallopian transfer (ZIFT) – also known as tubal embryo transfer is somewhat similar to
IVF wherein fertilization takes place outside the body. The young embryo is then inserted into the
fallopian tube and not the uterus.

4. Intracytoplasmic sperm injection (ICSI) – this technique is usually used for couples where the male
is infertile. Sometimes it is also used for those with failed IVF attempts or even for older couples.
Here the principle that is followed is that a single sperm is introduced into a mature egg, contrary to
what is seen in conventional fertilization techniques wherein the sperm and egg is placed in a petri
dish and the sperm fertilizes the egg on its own.

5. Artificial insemination – deliberate introduction of sperms into the female’s cervix or uterus to
achieve pregnancy without sexual intercourse.

Usage: ART is widely used by couples experiencing infertility, single individuals, and same-sex
couples wishing to have children.

Embryonic Stem Cells (ESCs)

ESCs are pluripotent stem cells derived from the inner cell mass of a blastocyst, an early-stage
pre-implantation embryo. Human embryos reach the blastocyst stage 4–5 days post fertilization, at
which time they consist of 50 – 150 cells.

Properties:

Embryonic stem cells (ESCs), derived from the blastocyst stage of early mammalian embryos, are
distinguished by their ability to differentiate into any embryonic cell type and by their ability to
self-renew. It is these traits that makes them valuable in the scientific and medical fields. ESCs have a
normal karyotype, maintain high telomerase activity, and exhibit remarkable long-term proliferative
potential.

1) Pluripotent:

Embryonic stem cells of the inner cell mass are pluripotent, meaning they are able to differentiate to
generate primitive ectoderm, which ultimately differentiates during gastrulation into all derivatives of
the three primary germ layers: ectoderm, endoderm and mesoderm.

These germ layers generate each of the more than 220 cell types in the adult human body.

When provided with the appropriate signals, ESCs initially form precursor cells that in subsequently
differentiate into the desired cell types.
Pluripotency distinguishes embryonic stem cells from adult stem cells, which are multipotent and can
only produce a limited number of cell types.

2) Self-renewal and repair of structure:

Under defined conditions, embryonic stem cells are capable of self- renewing indefinitely in an
undifferentiated state. Self-renewal conditions must prevent the cells from clumping and maintain an
environment that supports an unspecialized state.

Typically this is done in the lab with media containing serum and leukemia inhibitory factor or
serum-free media supplements with two inhibitory drugs ("2i"), the MEK inhibitor and GSK-3
inhibitor.

3) Growth:

ESCs divide very frequently due to a shortened G1 phase in their cell cycle.

Rapid cell division allows the cells to quickly grow in number, but not size, which is important for
early embryo development.

In ESCs, cyclin A and cyclin E proteins involved in the G1/S transition are always expressed at high
levels.

CDK2 that promote cell cycle progression are overactive, in part due to downregulation of their
inhibitors.

Retinoblastoma (Rb) proteins are hyperphosphorylated and inactivated in ESCs, leading to continual
expression of proliferation genes.

Although the shortened G1 phase has been linked to maintenance of pluripotency, ESCs grown in
serum-free 2i conditions do express hypo-phosphorylated active Rb proteins and have an elongated
G1 phase.

Uses:

Due to their plasticity and potentially unlimited capacity for self-renewal, embryonic stem cell
therapies have been proposed for regenerative medicine and tissue replacement after injury or
disease.

Pluripotent stem cells have shown promise in treating a number of varying conditions, including but
not limited to: spinal cord injuries, age related macular degeneration, diabetes, neurodegenerative
disorders (such as Parkinson's disease), AIDS, etc.

In addition to their potential in regenerative medicine, ESCs provide a possible alternative source of
tissue/organs. ESCs can also be used for research on early human development, certain genetic
disease, and in vitro toxicology testing.

Human embryonic stem cells have the potential to differentiate into various cell types, and, thus, may
be useful as a source of cells for transplantation or tissue engineering.