Potency

The term potency is sometimes used to mean the range of possible cell types or structures into
which a particular cell can develop.

Classification according to their potency:

Stem cells are categorized by their potential to differentiate into other types of cells.

Totipotent
Totipotency is the ability of a single cell to divide and produce all the differentiated cells
including extraembryonic tissues of an organism. Spores and zygotes are examples for totipotent
cells. Human development begins with a sperm fertilizes an egg to produce a single totipotent
cell (zygote) which in turn divides into identical totipotent cells in the first hours after
fertilization.[

Pluripotent

Pluripotent SC that has the potential to differentiate into any of the three germ layers: Ectoderm,
endoderm or mesoderm. E.g., ESCs isolated from inner cell mass (ICM) of blastocysts. Any fetal
or adult cell types can be derived from pluripotent SCs, but they lack the ability to contribute to
extraembryonic tissue like the placenta.

Multipotent
Multipotent progenitor cells have the potential to give rise to cells from multiple, but a limited
number of lineages. E.g., HSCs - A blood SC that can develop into several types of blood cells,
but lack the potential to develop into brain cells and other types of cells. Scientists have long
held the opinion that differentiated cells cannot be altered or induced to behave in any way other
than the way in which they have been naturally. In recent SC experiments, scientists have been
able to persuade blood SCs to behave like neurons or brain cells – A process known as
transdifferentiation.

Oligopotent
An oligopotent cell has the potency to differentiate into a few cell types. Examples of progenitor
cells are vascular SCs which have the capacity to become either endothelial or smooth
muscle cells.

Unipotent
A unipotent cell or precursor cell is one that has the capacity to differentiate into only one type of
cell/tissue type. The most common example of these cells in humans is skin cells. Some adult
stem cells, such as spermatogonia, are referred to as unipotent stem cells because they function
in the organism to generate only one cell type, the sperm cell in this example.
Fig: An example of the maturational series of stem cells. The differentiation of neurons is illustrated here
 Commitment

Commitment: Aspect of the intrinsic character of a cell or tissue region which causes it to
follow a particular pathway of development or fate.

Commitment of a cell or tissue region indicates that it is programmed to follow a particular

developmental pathway or fate.

Levels of developmental commitment

As cell fate becomes restricted following each decision in the developmental hierarchy, cells are
said to become committed to a certain fate (or developmental pathway). In animals, commitment
occurs in stages, initially reversible and then permanent (fig.). In plants, commitment appears
always to be reversible, so the following discussion relates only to animals.

Fig. .Stages of developmental commitment

An uncommitted cell can be described as naive, meaning that it has received no instructions
directing it along a particular developmental pathway. The fate of a cell is said to be specified if
the cell is directed to follow a certain developmental pathway and does so when placed in
isolation, which should provide a neutral environment. Specification may occur if a cell inherits
a particular cytoplasmic determinant or receives inductive signals from another cell. However,
that same cell placed in a different environment, such as in contact with other cells, may be
respecified by its interaction with those cells. This shows that the commitment at this stage is
reversible. The fate of a cell is said to be determined if it cannot be changed, regardless of the
cell’s environment. This shows that the commitment has now become irreversible.

Process of commitment

The process of commitment can be divided into two stages (Harrison 1933; Slack 1991).

1. Specification and 2. Determination

1. Specification: Commitment of a cell or tissue region which is manifested on culture in a
neutral medium but may still be reversible.

Or The fate of a cell or tissue is said to be specified when it is capable of differentiating

autonomously (i.e., by itself) when placed in an environment that is neutral with respect to the
developmental pathway, such as in a petri dish or test tube.

Or A group of cells is called specified if, when isolated and cultured in the neutral environment
of a simple culture medium away from the embryo, they develop more or less according to their
normal fate.

For example, cells at the animal pole of the amphibian blastula are specified to form ectoderm,
and will form epidermis when isolated. Cells that are ‘specified’ in this technical sense need not
yet be ‘determined’, for influences from other cells can change their normal fate. If tissue from
the animal pole is put in contact with cells from the vegetal pole, some animal pole tissue will
form mesoderm instead of epidermis. At a later stage of development, however, the cells in the
animal region have become determined as ectoderm and their fate cannot then be altered. Tests
for specification rely on culturing the tissue in a neutral environment lacking any inducing
signals, and this is often difficult to achieve.

2. Determination

The second stage of commitment is determination.

A cell or tissue is said to be determined when it is capable of differentiating autonomously even

when placed into another region of the embryo or a cluster of differently specified cells in a petri
dish. If a cell or tissue type is able to differentiate according to its specified fate even under these
circumstances, it is assumed that commitment is irreversible.

Or It is the type of developmental commitment of cells or tissue explants that is irreversible

following grafting to any different location in the embryo.

Determination implies a stable change in the internal state of a cell, and an alteration in the
pattern of gene activity is assumed to be the initial step, leading to a change in the proteins
produced in the cell. Different mesoderm cells, for example, eventually become determined as
muscle, cartilage, bone, the fibroblasts of connective tissue, and the cells of the dermis of the
skin.
An important concept in the process of cell determination is that of lineage, the
direct line of descent of a particular cell. Once a cell is determined, this stable change
is inherited by its descendants. Thus, the lineage of a cell has important consequences
in terms of what type of cell it can eventually become. For example, once determined
as ectoderm, cells will not subsequently give rise to endodermal tissues, or vice versa
The state of determination of cells at any developmental stage can be demonstrated
by transplantation experiments. At the gastrula stage of the amphibian embryo, one
can graft the ectodermal cells that give rise to the eye into the side of the body and
show that the cells develop according to their new position; that is, into mesodermal
cells. At this early stage, their potential for development is much greater than their
normal fate. However, if the same operation is done at a later stage, then the cells of
the future eye region will form structures typical of an eye (Fig. ). At the earlier stage the cells
were not yet determined as prospective eye cells, whereas later they
had become so, and so the graft development was autonomous—that is, it developed
as eye irrespective of its new location.

Fig. Determination of the eye region with time in amphibian development. If the region of the gastrula that will normally
give rise to an eye is grafted into the trunk region of a neurula (middle panel), the graft forms structures typical of its new
location, such as notochord and somites. If, however, the eye region from a neurula is grafted into the same site (bottom panel), it
develops as an eye-like structure because at this later stage it has become determined.

It is a general feature of development that cells in the early embryo are less narrowly determined
than those at later stages; with time, cells become more and more
restricted in their developmental potential. We assume that determination involves
a change in the genes that are expressed by the cell, and that this change fixes or
restricts the cell’s fate, thus reducing its developmental options.
In summary, then, during embryogenesis an undifferentiated cell matures through
specific stages that cumulatively commit it to a specific fate: first specification, then
determination, and finally differentiation. During specification, there are three major
strategies that embryos can exhibit: autonomous, conditional, and syncytial. Embryos
of different species use different combinations of these strategies.

Autonomous Specification :

In this type of specification the blastomeres of the early embryo have a set of critical
determination factors called cytoplasmic determinants that they get from the egg cytoplasm.
The cytoplasm is not homogenous but has different regions containing cytoplasmic determinants
that are often transcription factors or mRNA encoding factors that will influence the cell’s
development. There is asymmetric segregation of cellular determinants during cell division.
Thus, some cells will have more or less of the transcription factors and some may have none.
This will result in daughter cells with different cell fates. These cells already “know” what they
are fated to become.
Some of the best examples of autonomous specification are seen in the
development of molluscs, annelids and tunicate embryos (Fig.). In the tunicate embryo
experimental results showed that when the blastomeres were separated at the 8 cell stage each
blastomere gave rise to the respective cell type it was fated to make. Moreover, if some of
the cytoplasmic determinants like those for muscle development were removed and placed in
another cell that cell began to differentiate into muscle cells. Such embryos that have their fates
specified early in cleavage are known as mosaic embryos and their development is known
as mosaic development.

Fig: Autonomous specification seen in tunicate embryo. The cells were separated at the 8 cell stage and each
cell gave rise to only those structures that it would have made in the entire embryo.
Conditional specification: The ability of cells to achieve their respective fates
by interactions with other cells. What a cell becomes is in large measure specified by paracrine
factors secreted by its neighbors.

Here, what a cell becomes is specified by the array of interactions it has with its neighbors,
which may include cell-to-cell contacts (juxtacrine factors), secreted signals (paracrine
factors), or the physical properties of its local environment (mechanical stress).

Various experiments conducted by embryologists since the 1890s that dealt with the separation
of early blastomeres in sea urchin which resulted in each blastomere giving rise to a separate
embryo (recall both Roux’s and Dreisch’s experiments). Thus, it was seen that each blastomere
or cell acquires its identity based on its position and its interaction with its neighbouring cells
and molecules with which it comes in contact (Roux’s experiment) and when the early
blastomeres were separated they lost that interaction and were able to develop into separate
embryos (Dreisch’s experiment).Embryos, especially vertebrate embryos in which the early
blastomeres are conditionally specified were traditionally called regulative embryos.

Fig: Conditional specification. a) The fate of a cell depends on its position in the embryo; b) the region from
where the cell had been removed compensates for the loss and the embryo develops normally

Syncytial specification: The interactions of nuclei and transcription factors,

which eventually result in cell specification, that take place in a common cytoplasm, as in the
early Drosophila embryo.

Insects employ a specialized strategy, syncytial specification, where the egg nucleus divides a
number of times to generate a syncytium containing thousands of nuclei. Cell fates are
determined prior to cellularization by interactions between diffusible regulators and individual
nuclei. Drosophila development is illustrative of this strategy.
The distribution of maternal mRNAs in the Drosophila egg is critical in the establishment of the
anteroposterior axis. Maternal bicoid mRNA is located at the anterior pole and maternal nanos
mRNA is located at the posterior pole. The proteins encoded by these messages control
the establishment of gradients of transcription factors in the early syncytial
embryo, which in turn control downstream genes that divide the embryo into
segments and confer upon each segment its positional identity.
 Embryonic Induction

One of the most remarkable features of embryonic development is the precision with which
developmental signals are generated, transmitted and received. These signals are of many types,
and their effect may be made manifest in a variety of ways. One of the most important varieties
of embryonic signal calling is embryonic induction.

By Induction we mean an effect of one embryonic tissue (The inductor) on another, so that the
developmental course of the responder tissue is qualitatively changed from, what it would have
been in the absence of inductor. i.e The process by which one cell population influences the
development of neighboring cells via interactions at close range. It usually involve intercellular
signaling events, which lead to the up regulation of different combinations of developmental
control genes in each zone of cells

Definition: The process whereby the development of one group of cells, called the competent
region, is altered by an inducing factor from another group, called a signaling center or
organizer.

Primary embryonic induction: The process whereby the dorsal axis and central nervous
system forms through interactions with the underlying mesoderm, derived from the dorsal
lip of the blastopore in amphibian embryos.

Reciprocal inductions: One tissue induces another, and that tissue then acts back on the
original inducing tissue and induces it, thus the inducer becomes the induced

Inductive interactions occur throughout much of embryonic development and even into postnatal
life. For e.g. In vertebrate, the first major inductive event involves the induction of mesoderm in
embryonic cleavage. This is followed by the induction of the nervous system (often called
primary induction) during the shortly after gastrulation.

The nervous system itself then induces other structures (often sensory organ) in what sometimes
called secondary inductions, and the tissues produced through secondary induction sometimes
induces the formation of other structures through tertiary inductions. The formation of internal
organ almost all internal organs occurs through inductive interactions.(Fig 1-20)
Let us understand this phenomenon by taking the example of the development of the vertebrate
eye. When the development of eye is initiated, two bulges are seen in the brain that approach the
surface ectoderm in the head region. The head ectoderm is competent to respond to the paracrine
factors released by the brain bulges that are the optic vesicles. The head ectoderm cells are
induced to form the lens tissue of the eye that is, the genes for expressing the lens protein are
activated. The prospective lens cells in turn secrete another paracrine factor that instructs the
optic vesicle to form the retina. Note that the two cell types that co-construct the eye induce each
other. The important part is that the head ectoderm is the only region in the embryo that is
competent to respond to the instructions from the optic vesicle signals.
Types: Inductive interactions have been classified as permissive or instructive (Slack 1993).

1. Permissive induction: In permissive induction an inductive signal is required to bring about

the development of a specific structure (e.g. metanephric kidney). The responding tissue will
form no other structure, even through artificial inductive stimuli are applied. In the absence of
permissive induction, the responding tissue fails to develop into any structure.

2. Instructive induction: An instructive induction is one in which the responding tissue has the
option of forming more than one type of tissue, depending upon the nature of the inductive
stimulus. For e.g. without appropriate induction, cells that would have normally formed
mesoderm in the early amphibian embryo go on to form ectoderm.
Similarly in absence of embryonic lens, the cells that would have formed the corneas
differentiate into epidermis.
Instructive and permissive induction can be distinguished by grafting experiments. For example,
in mammals, the cardiogenic mesenchyme (future heart) is required to induce hepatocyte
development in the presumptive liver-forming region of the foregut. The signals could be
instructing the foregut to form hepatocytes instead of other cell types or could be permitting the
differentiation of hepatocyte cells that have already been specified by other mechanisms. If the
cardiogenic mesenchyme is grafted under the hindgut, no hepatocytes are induced. The inductive
signals from the cardiogenic mesenchyme are therefore permissive rather than instructive.

Transmission of Inducing signals between cells

Induction can occur over different ranges

In the most extreme case, endocrine signaling involves the release of inductive signals by
one type of cell located a considerable distance from the responding cell population, and the
molecule must travel to its target through the vascular system. This is the mechanism
utilized by many hormones.
Most inductive interactions in early development occur locally. This can involve the release
of a diffusible substance (paracrine signaling) or a substance that is deposited in the
extracellular matrix.
The range of such signals varies, but rarely exceeds a few cell diameters. The range of a
signal can be extended by a relay mechanism, in which one signal causes the responding
cell to release another signal, which similarly induces a third and so on. It is therefore
possible for the responding tissue to take on the inductive properties of the inducer, a
phenomenon termed homeogenetic induction
In other cases, the signal may only reach the cell adjacent to the inducer, particularly in
cases where direct contact between cells is required because the inductive signal is fixed to
the cell membrane, or it travels through gap junctions (juxtacrine signaling). Cells may
also reciprocally induce each other, i.e. each cell acts as the inducer of the other, and
responds to the other’s inducing signal. Such reciprocal induction occurs in the kidney,
where the ureteric bud induces the metanephric blastema to differentiate into the adult
kidney structures, while the blastema induces the ureteric bud to proliferate and branch.
Fig: Ranges and types of induction
 Competence

Competence is a property of the cell responding to induction. A cell is described as competent if

it can respond to the inductive signal by undergoing all the appropriate molecular changes that
allow it to follow the induced’ developmental pathway. In the absence of induction, the cell
eventually becomes determined to an alternative pathway, and this coincides with its loss of
competence to respond to induction.

In the case of endocrine or paracrine signaling, competence depends on the synthesis of all the
components of the signal transduction pathway that link the inductive signal to its target, such as
a transcription factor, in the responding cell. If any of these components is lost,e.g. the cell
surface receptor, the signal transduction apparatus, or the down- stream target transcription factor
itself, the cell loses competence.

In the case of juxtracrine signaling, a cell may also lose competence simply by breaking contact
with the inducing cell. This may reflect the movement of cells away from each other, or the
disassembly of gap junctions.

Genomic equivalence

Definition :

The theory that every cell of an organism has the same genome as every other cell.

Explanation:

With very few exceptions, all differentiated cells in a given organism contain the same DNA,
which is the same as the DNA in the fertilized egg. This means that almost all differentiated cells
in principle contain all the information required to generate a complete new individual. Evidence
for genomic equivalence comes from molecular analysis including Southern blot hybridization
and PCR amplification of genomic DNA isolated from embryos and various adult tissues. When
probed for different developmental marker genes and cell type- specific genes, all banding
patterns and PCR products are the same.

Evidence for Genomic Equivalence

Ian Wilmut and colleagues took cells from the mammary gland of a 6-year-old pregnant ewe and
placed them in culture ( Wilmut et al. 1997). The culture medium was formulated to keep the cell
nuclei at the intact diploid stage (G1) of the cell cycle; this cell-cycle stage turned out to be
critical. The researchers then obtained oocytes from a different strain of sheep and removed their
nuclei. These oocytes had to be in the second meiotic metaphase, the stage at which they are
usually fertilized. The donor cell and the enucleated oocyte were brought together, and electric
pulses were sent through them, thereby destabilizing the cell membranes and allowing the cells
to fuse. The same electric pulses that fused the cells activated the egg to begin development. The
resulting embryos were eventually transferred into the uteri of pregnant sheep

Of the 434 sheep oocytes originally used in this experiment, only one survived: Dolly. DNA
analysis confirmed that the nuclei of Dolly’s cells were derived from the strain of sheep from
which the donor nucleus was taken (Ashworth et al. 1998; Signer et al. 1998). Cloning of adult
mammals has been confirmed in guinea pigs, rabbits, rats, mice, dogs, cats, horses, and cows. In
2003, a cloned mule became the first sterile animal to be so reproduced (Woods et al. 2003).
Thus, it appears that the nuclei of vertebrate adult somatic cells contain all the genes needed to
generate an adult organism. No genes necessary for development have been lost or mutated in
thesomatic cells; their nuclei are equivalent.
 Morphogen
A morphogen is a substance that can influence cell fate, but has different effects
at different concentrations.
Or
Morphogens Greek, “form-givers.” These are diffusible biochemical molecules that can
determine the fate of a cell by their concentrations, in that cells exposed to high levels of a
morphogen will activate different gene
In its simplest form, the morphogen model suggests that positional information along an axis can
be generated by the synthesis of a morphogen at a source at one end of the axis, and diffusion
away from the source would set up a morphogen gradient. Cells along the axis would receive
different concentrations of the morphogen and this would induce different
patterns of gene expression at different concentration thresholds (Fig. ).

Fig. Principle of a morphogen gradient

Such concentration-dependent patterns of gene expression would represent the ‘address’ or

positional identity of the cell. A number of systems have been studied that appear to involve
morphogen gradients.

Examples

In early Drosophila development, the Hunchback protein is distributed in a gradient along

the anteroposterior axis of the syncytial embryo. Hunchback is a transcription factor and
activates downstream genes such as Kruppel, knirps and giauf in a concentration dependent
manner. The latter genes are expressed in stripes defining broad sectors of the anteroposterior
axis.

Similarly in vertebrate limb development, the signaling protein Sonic hedgehog is

expressed in the posterior zone of polarizing activity (ZPA) of the limb bud, and establishes a
concentration gradient running from posterior to anterior. Although the mechanism is complex
the result is that different HoxD genes are activated in concentric rings surrounding the ZPA,
resulting in the formation of different digits from essentially the same cell types.
In both these systems, interfering with the level and /or position of the morphogen has
profound effects on development. The data suggest that such experimental manipulations cause
cells to receive the wrong positional information; consequently they adopt the wrong positional
identities, resulting in the appearance of regionally inappropriate structures.
In the early Drosophila embryo, transcription factors can act as morphogens by diffusing freely
through the cytoplasm to establish concentration gradients.

Similarly, the lipid signaling molecule retinoic acid may be able to establish a
concentration gradient across a field of cells by free diffusion. It is thought
unlikely, however, that secreted proteins of the Hedgehog, Wingless /Wnt and
Decapentaplegic/ BMP families, all of which function as morphogens in a
variety of developmental systems, can establish concentration gradients in this
manner.
Mechanism of action
Recent evidence has emerged to suggest that such proteins could act as
morphogens in a variety of ways (Fig. ):

• Initiating a signal relay, where one signaling protein diffuses over a short range, but
induces adjacent cells to produce another signaling protein, which acts in the same way, with
diminishing effect across a field of cells. For example, Sonic hedgehog has been shown to induce
BMP2 expression in adjacent cells in the vertebrate limb.
• Responding cells could extend cytoplasmic processes (cytonemes) to contact the inducing
cell, and the longest cytonemes would be extended by more distant cells. Ligands binding to
receptors on the end of the cytonemes would induce the same signal transduction pathway in all
cytonemes, but the response to signaling might be diminished as a function of increased
distance front the cell nucleus. Such cytoplasmic processes have been observed in Drosophila.
 • The signaling protein could be propagated by a carrier. Proteoglycans in the
extracellular matrix have been implicated in this role, and mutations
affecting proteoglycan synthesis such as sugarless and tonI refs have been
shown to inhibit Hedgehog and Wingless signaling in Drosophila.

• The signaling protein could be transported from cell to cell by cyclical endocytosis and
exocytosis. Evidence for the intracellular transport of Wingless has been reported in Drosophila.
Finally, it should be noted that concentration-dependent induction can also be
established by an ‘anti-morphogen’ gradient. For example, a morphogen can be secreted
uniformly, but the level of its activity can be graded by the diffusion of an inhibitor. This is
apparent in systems such as the vertebrate organizer, which secretes molecules that inhibit BMPs
and Wnts. Similarly, differential responses to a uniform concentration of inducer can be achieved
by varying the levels of the receptor in the responding cells. However, this requires pre- existing
polarity in the system.

Difference between morphogen and morphogenesis

Morphogenesis is the process by which structures form during development, reflecting different
types of cell behavior. A morphogen is an inducer that elicits different responses at different
concentrations; the term literally means ‘form-generating substance’.
 Differentiation

It is the process by which a cell undergoes a progressive, coordinated change to a more

specialized cell type, brought about by large-scale changes in gene expression.
Or
The generation of specialized cell types is called differentiation, a process during which a cell
ceases to divide and develops specialized structural elements and distinct functional properties.
Cell differentiation has a twofold meaning.
The term may mean: (1) cells becoming different from each other, that is, the divergence of
developmental pathways taken by the various cell types;
or (2) cells becoming different in time: each cell takes a cell-type-specific pathway until it
reaches maturity and becomes terminally differentiated. The development of each cell begins
with a pluripotent founder cell-at the outset, the fertilized egg which is capable of dividing and
gives rise to several cell types. An act of decision, called determination or commitment, directs
the developmental path of the descendants to different goals.
A cell that has become committed (programmed) to following a distinct pathway is designated
by the suffix -blast (e.g., neuroblast, erythroblast). Such a committed cell may retain the ability
to divide. After some rounds of cell division, the derivatives of the founder cell proceed to
undergo terminal differentiation. They acquire their specific molecular equipment, specific form,
and function. In so doing, the cells will, as a rule, lose their ability to divide. The suffix -cyte
indicates that this has happened.
e.g. Skeletal muscle is the voluntary muscle, making up a large proportion of the mass of a
vertebrate animal. It consists of multinucleate myofibers, which contain myofibrils composed of
the contractile proteins actin and myosin. Differentiation of skeletal muscle involves first the
commitment of cells to become myoblasts, then a period of multiplication and/or migration, then
a cessation of division and fusion of the myoblasts to form the multinucleate myofibers. In
normal development the cessation of proliferation occurs after myoblasts have migrated from
dermomyotome to the underlying myotomes. Once postmitotic, they should strictly be called
“myocytes,” but the name “myoblast” still persists.

Fig: Key features of the differentiation of vertebrate skeletal muscle.