CREDIT 04- UNIT 03: CONTROL OF DEVELOPMENT

LEARNING OBJECTIVES
After studying this unit, you should be able to
 Differentiate between Embryology and Developmental Biology
 State the progressive stages in development of a multicellular organism;
 Describe the features of the various nonchordate and chordate animals used as
model organisms.
 Explain the role of fate maps and patterns of development;
 State the concepts of cell specification, determination and differentiation.
 Explain the concept of genomic equivalence, totipotency and pluripotency
through classic experiments of Briggs and King.
 Discuss the mechanism of differential gene expression.

INTRODUCTION
Key to multicellularity is the coordinated interaction of the various cells that make up
the body. Indeed, patterning of embryos, establishment of cell type diversity, and
formation of tissues and organs all rely on cell -to-cell communication during
development. Thus, arguably one of the most important principles of developmental
biology involves “one group of cells changing the behavior of an adjacent set of cells,
causing them to change their shape, mitotic rate, or fate”

04-03-01: FUNDAMENTAL PROCESSES IN DEVELOPMENT

The plan for an organism's growth, starting from a fertilized egg to an adult , is
stored in two main things: (i) the genes in the fertilized egg, and (ii) some special clues
called cytoplasmic determinants in the egg's cytoplasm. As the organism develops, the
cells in the fertilized egg transform into many types and work together t o make the
different parts of the body. They communicate and coordinate to build a complete and
functional organism. A key idea for all living things is that the fertilized egg takes time
to go through different stages of development before becoming a full y formed and
functional organism.
The stages of development that occur between fertilization and the birth of an organism
(Fig.4.3.1) are collectively known as embryogenesis. It is during embryogenesis that
the genotype (genes of the developing organism) of the organism determines the
morphological appearance (phenotype) of the organism. Each animal whether it is a
fruit fly or earthworm or frog, or bird or a mammal undergoes the same basic stages of
development which include:
i) Fertilization - Combining Special Cells: This is when the mature cells from the
mom and dad join together. Each of these cells has only half of the special instructions

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(chromosomes) needed. When they come together, they make a new cell called a
zygote, and this cell has all the necessary instructions to grow into a new organism that
looks similar to its parents.
ii) Cleavage or Fast Cell Division - Splitting Up: After fertilization, the zygote starts
dividing quickly into many small cells called blastomeres. These cells don't grow
bigger between divisions, so they become smaller with each split. They form a cluster
of cells known as morula and then a blastula during this phase.
iii) Gastrulation - Shaping Up: At this stage, the cell division slows down, and the
blastula cells start moving and rearranging. This causes the creation of three layers -
ectoderm, mesoderm, and endoderm. These layers work together to form the different
parts of the body.
iv) Morphogenesis- Giving Shape: This is when the cells in the embryo start to
become different from each other. It's like the process of giving the embryo its specific
shape.
v) Organogenesis - Building Organs: Now, the organs start forming, and the embryo
becomes a functional little being. At this point, you can clearly see that it belongs to a
particular species. This is when the new individual is born and begins its life, growing
and living until the end.
In many species a group of cells are set aside and do not pa rticipate in the formation of
the embryo. These are known as germ cells and are used to produce the next generation.
All other cells of the body are known as somatic cells. The germ cells migrate in the
embryo to form the gonads and give rise to gametes in the adult organism. However, the
process of development does not stop at birth, it is seen as metamorphosis and
regeneration in some animal groups and finally as aging or senescence

Fig. 4.3.1 we have shown the life cycle of frog as an example depicting all the
above-mentioned stages of development.

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HISTORICAL BACKGROUND OF DEVELOPMENTAL BIOLOGY
The exploration of animal development traces back to the 4th century BC when
simple observations of visible eggs and embryos began. Aristotle, a pioneer in this
field, documented variations in the life cycles of animals. He classified animals based
on their methods of birth, such as oviparity (egg -laying), viviparity (direct birth of
young in mammals), and ovoviviparity (eggs hatching within the body).
Aristotle also observed two types of division patterns during cleavage in fertilized eggs:
holoblastic division (entire embryo divides) found in frogs and mammals, and
meroblastic division (only a part divides, the rest nourishes the embryo) seen in chicks.
Based on his studies on chick embryos, Aristotle introduced the theory of epigenesis,
proposing that new structures develop progressively during embryonic development.
According to this theory, there are no preformed tissues or organs at the start of
development.However, another theory gained popularity later —the theory of
preformation. According to preformationists, all embryo parts were preformed and
simply enlarged over time. They believed in a preformed miniature, the homunculus,
either in sperm (spermists) or eggs (ovists). Embryology saw little progress until 1651
when William Harvey made a groundbreaking observation of the blastoderm in a chick.
He concluded that all animals arise from eggs, marking a significant advancement in
our understanding of embryonic development. Harvey also observed the formation of
red dots of pooled blood preceding the development of the heart and vessels.
The debate between the two theories, preformation and epigenesis, persisted until the
17th and early 18th centuries. Kasper Freidrich Wolff, building on Aristotle's
observations, supported epigenesis, challenging the notion of preformation. He studied
chick embryos, demonstrating that new tissues, like the heart, intestine, and blood cells,
form during development and are not perfor med in miniature.
The final blow to the preformation theory came in the 1820s with the emergence
of the cell theory. This theory, coupled with advanced staining techniques and improved
microscopy, paved the way for descriptive embryology. Christian Pander (1794-1865)
identified three germ layers (ectoderm, mesoderm, and endoderm) in chick embryos,
highlighting their interdependence. Martin Rathke (1793 -1860) discovered similar
layers in crayfish, proposing that three germ layers were not exclusive to verteb rates
but also found in invertebrates.
Embryologists studying vertebrate embryos observed striking similarities across
different groups. In 1828, Karl Ernst Von Baer proposed this as evidence of evolution
and formulated his four laws of animal development. Von Baer's work, outlined in his
book "On the Development of Animals," published in 1828 and 1837, reviewed
vertebrate development, laying the foundation for our understanding of embryology.
These laws, translated by Thomas Henry Huxley in Scientific Memo irs are asfollows:

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 1. In embryos, the features common to a big group show up first, followed by the
more specific traits.
2. The development process goes from the most common features to the less
common ones until the very specific ones appear.
3. Instead of going through various forms, each animal's embryo becomes distinct
from others.
4. Essentially, the embryo of a higher form doesn't look like the embryos of other
forms but only like its own.
ANIMAL MODELS IN DEVELOPMENTAL STUDIES
In the 20th century, the field of experimental embryology witnessed significant
progress, revealing striking similarities in the developmental mechanisms across
various animals. These similarities extended not only to genes and proteins but also
encompassed entire signaling pathways and their responses during embryogenesis,
indicating a high degree of conservation throughout evolution. Researchers delving into
gene expression experiments found that fundamental mechanisms of development were
controlled by a common set of regulatory genes, orchestrating distinct functions in
developing embryos.Several model organisms played crucial roles in advancing
developmental studies. Sea urchins, echinoderms found in oceans, emerged as key
model systems. Their suitability lay in the ease of obtaining gametes, visibility of large
eggs, synchronous development, and transparency of embryos, making them ideal for
microscopic observation, gene expression, and genetic analysis.
Amphibians, such as frogs like Xenopus levisand Rana pipens, offered advantages with
their large, observable eggs laid in water. The accessibility of these organisms, coupled
with their ease of breeding in captivity and short development times, facilitated
significant insights into molecular mechanisms go verning embryonic cell division and
differentiation.
Drosophila melanogaster, the fruit fly, became a favored model for genetic
analysis in the mid-20th century. Its attributes, including ease of cultivation, short
reproductive cycle, and a genome containing approximately 15,500 genes, contributed
to its significance in developmental biology. Mutants of Drosophila were particularly
valuable in understanding gene functions.
Caenorhabditis elegans, a free-living nematode, provided a simple and
transparent system for developmental studies. Its fully sequenced genome and invariant
cell lineage allowed scientists to work out a detailed cell development lineage.
Zebrafish (Danio rerio) emerged as a newer model organism, combining
features like small size, robustness, cost-effectiveness, large transparent eggs, and rapid
embryonic development. The zebrafish's fully sequenced genome and its sharing of 70%
of genes with humans made it an ideal candidate for molecular studies and
understanding early development.Over ti me, developmental biology moved from

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studying big-picture embryonic processes to diving into the tiny details of cells and
molecules. Improved tools and techniques enabled scientists to explore the genetic and
molecular foundations of animal development mo re profoundly. As this field
progresses, selecting the right model organisms becomes crucial, taking into account
factors like accessibility, ease of manipulation, and compatibility with experimental
methods. Animal developmental biology, changing and grow ing, now sits at the exciting
crossroads of genetics and embryology.
EMERGENCE OF PATTERNS
As embryonic development proceeds it involves the emergence of a pattern
which shows the overall position of cells in the embryo so that the body’s shape and
form begins to emerge. This is followed by cell differentiation and growth.
Development is a gradual process by which a complex multicellular organism arises
from a single cell (the zygote). It involves 5 major overlapping processes:
1) Growth = Increase in Size
2) Cell Division= Increase in Number
3) Differentiation = Diversification of Cell Type s
4) Pattern Formation = Organization
5) Morphogenesis = Generation of Shapes and Structures
PATTERN F ORMATION
In the journey from a single cell (zygote) to a developed organism, cells undergo a
remarkable transformation. This process, known as patter n formation, is like a blueprint
that guides initially similar cells to take on distinct roles based on where they are in the
growing embryo. It's how a well-organized structure emerges during
development.Simply put, pattern formation in developmental biol ogy is the way cells in
a developing embryo, initially alike, shape into intricate forms and functions based on
their positions. The future identity of a cell hinges on where it is located within the
growing embryo. Picture it as a step-by-step process where a cell gradually determines
its final role.Consider the development of the face as an example. During this process,
cells follow a pattern to figure out where the nose, ears, and eyes should form, and how
the muscles should support the face's structure. In the animal world, pattern formation
sets in early during embryonic development, achieved through various cellular and
molecular mechanisms at different times in different organisms.Pattern formation kicks
off with a crucial step—establishing the body axes of an animal. Envision drawing two
lines on the creature: one stretching from the head to the tail (anterio -posterier axis),
and the other extending from the back to the underside (dorso -ventral axis). Many of
the animals we're exploring in this course showcase bilateral symmetry, where the left
and right sides mirror each other. Picture these axes intersecting at right angles,
creating a coordinate system that plays a fundamental role in specifying the animal's

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form. This initial setup becomes the foundation for shaping the creature as it develops,
setting the stage for further intricate patterns to emerge.

Fig.4. 3.2: The main axes in a developing embryo of Xenopus laevis

In vertebrate animals, although external symmetry is evident, internal organs exhibit
asymmetry; the heart resides on the left, while the liver occupies the right. The
establishment of axes in the embryo is an early developmental process involving
several genes. Simultaneously, before the external body axes manifest, the egg
possesses a distinct polarity. As these axes take shape, cells in triploblastic animal
embryos organize into three germ layers: ectoderm (external), mesoderm (middle), and
endoderm (internal). In diploblastic animals, embryos organize cells into two germ
layers: ectoderm (external) and endoderm (internal). During the gastrula stage, the
positions of cells in the germinal layers, in both diploblastic and triploblastic embryos,
foreshadow their prospective fate by the end of development. Further in the pattern
formation process, cells within these layers undergo differentiation, assuming diverse
identities to give rise to various tissues and organs like skin, muscle, blood cells, and
neural cells.
Pattern formation in embryonic development can occur through two distinct
mechanisms. Firstly, there are cell-to-cell interactions, also termed inductive
mechanisms, where certain cells induce or instruct others to alter their behavior and
develop into different cell types. Secondly, morphogenetic mechanisms act on
established patterns to shape three-dimensional tissues and organs, involving changes in
cell location without altering their behavior. For example, during gastrulation in frogs,
the endoderm and mesoderm migrate inside the embryo from the surface, leading to the
formation of a tube-within-a-tube structure for the gut. Throughout development, the
embryo undergoes notable changes in appearance or morphology, driven by extensive
cell migration that also induces alterations in the embryo's shape. Additionally,
programmed cell death is part of this process.
SPECIFICATION
Embryos of different animal species show different strategies of specification
Autonomous specification is a type of cellular determination observed in early
embryos, where blastomeres possess critical determination factors known as

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cytoplasmic determinants obtained from the egg cytoplasm. The cytoplasm is not
uniform but consists of distinct regions containing these determin ants, often
transcription factors or mRNA encoding factors that impact cell development. During
cell division, there is an uneven distribution of these determinants, leading to daughter
cells with varying amounts of transcription factors or none at all. Th is results in cells
having predetermined fates. Autonomous specification is well exemplified in the
development of molluscs, annelids, and tunicate embryos. In tunicate embryos,
experiments separating blastomeres at the 8 -cell stage revealed that each blastomere
gave rise to the respective cell type it was fated to produce. Additionally, if certain
cytoplasmic determinants, like those for muscle development, were removed and
transferred to another cell, that cell started differentiating into muscle cells. E mbryos
with early-specified fates during cleavage are termed mosaic embryos, and their
development is referred to as mosaic development.
Conditional specification: in this process the cells achieve their final fate by
interacting with other cells or their local environment. These cells – to cell interactions
may be of various kinds (which we will discuss in greater detail in the next unit). The
fate of a cell in this type of specification depends on its position in the embryo. For
example, if the cells from a region of a vertebrate blastula ‘A’ that are known to give
rise to the dorsal region of the embryo are taken out and transplanted to the region of
the blastula ‘B’ that gives rise to ventral region in the embryo, then A type of cells will
change their fate and differentiate into B type cells. Moreover, the region from where A
type cells were taken will also continue to develop normally (Fig. 4.3.4).

Fig. 4.3.3: Conditional specification - a) Cell fate depends on embryo position; b)

Removal-compensated region ensures normal development.

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,

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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. With further use of
molecular biology in the process of development it has beenSyncytial specification: is
the third strategy which is seen mostly during embryo development of insects and has
been demonstrated best in the Drosophila embryo. The cytoplasm in Drosophila egg
already has certain factors or maternal determinants distributed in it unequally. For
instance, the anterior most part of the egg contains an mRNA called Bicoid mRNA that
encodes a protein called Bicoid. The posterior most part of the egg contains an mRNA
called Nanos mRNA that encodes a protein called Nanos. After fertilization of the egg,
these two mRNAs are translated into their respective proteins in the cytoplasm,
contributed by the ovum. During the early cleavage stage the nucleus alone divides and
the cytoplasm does not separate or undergo cellularization to form individual cells.This
results in formation of the blastoderm (a blastula having the form of a disc of cells on
top of the yolk) which in Drosophila appears in the form of a large cell with many
nuclei and a common plasma membrane (Fig.4.3.5).

Fig. 4.3.4: Syncytial specification in Drosophila melanogaster - Egg has

concentrated Bicoid mRNA in anterior and Nanos mRNA in posterior. After
fertilization, cytoplasm contains both mRNAs, influencing noncellularized nuclei
based on the Bicoid and Nanos protein concentration gradients.
A cytoplasm that contains many nuclei is called a syncytium as you may recall.
It is in this syncytial blastoderm that the cell i dentities are determined even before
individual cells are formed. Research has shown that the concentration of Bicoid
protein is highest in the anterior part of the cytoplam of the embryo and declines toward

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the posterior end. While the concentration of th e Nanos protein is maximum in the
posterior part of cytoplasm of embryo and declines as it diffuses anteriorly. Thus, the
long axis of the Drosophila egg is spanned by two opposing gradients —one of Bicoid
protein coming from the anterior side, and one of N anos protein coming from the
posterior side. The Bicoid and Nanos proteins form a coordinate system based on their
ratios, such that each region of the embryo will be distinguished by a different ratio or
concentration of the two proteins. As the nuclei di vide and enter different regions of the
egg cytoplasm, they will be instructed by these ratios as to which position should they
occupy along the anterior-posterior axis of the animal. Those nuclei in regions
containing high amounts of Bicoid and little Nan os will be instructed to activate those
genes necessary for producing the head. Those nuclei in regions with slightly less
Bicoid but with a small amount of Nanos will be instructed to activate those genes that
generate the thorax. Those nuclei in regions that have little or no Bicoid and plenty of
Nanos will be instructed to form the abdominal structures. After the 13th cleavage
division just before gastrulation the cell membranes are laid down to separate the
nuclei. The nucleus is able to keep its positi on in the embryo because of its exposure to
the different concentrations of the cytoplasmic determinants and each cell or blastomere
after cellularisation would know what part of the embryo it will become. Thus, the
specific fate of each cell is determined both autonomously (from cytoplasmic
determinants) and conditionally (by interaction with other neighboring cells).

MULTIPLE CHOICE QUESTIONS (MCQs):

MCQ 01: During embryogenesis, which process involves the movement and
rearrangement of blastula cells to form three distinct layers?
a) Fertilization
b) Cleavage
c) Gastrulation
d) Morphogenesis
Answer: c) Gastrulation
MCQ 02: Which historical figure introduced the theory of epigenesis, proposing
that new structures develop progressively during embryonic development?
a) Aristotle
b) William Harvey
c) Kasper Freidrich Wolff
d) Karl Ernst Von Baer
Answer: c) Kasper Freidrich Wolff
MCQ 03: Which model organism is particularly valuable in understanding gene
functions due to its mutants?
a) Sea urchin

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other cells of the body? Or why insulin hormone is produced and secreted only from
certain cells of the pancreas and never in the kidneys or in the brain? By 1960s, based
on experimental evidence, the concept of differential gene expression came into
existence to provide answer to these and several other questions and to explain how
similar looking cells with the same type of genetic complemen t differentiate into
different types of cells.
Genomic Equivalence
The best test of whether all somatic cells contain the same complement of genes
as the fertilized egg from which they have arisen is to check if the nuclei of the
differentiated somatic cells still have the ability or potency to generate all types of
cells. If indeed this is the case, then a nucleus taken from one type of cell in the body
should be totipotent (having the ability to produce all types of cells) and if transplanted
into an activated enucleated (nucleus removed) cell should be able to give rise to all the
cells of the body.
In the 1950s, Robert Briggs and Thomas King conceptualized and conducted the
experiments for determining the totipotentency of the nucleus of early embryo nic cells.
In their experiment they first combined the technique of enucleation and activation of
an oocyte of the leopard frog Rana pipens. The oocyte was activated by pricking it with
a sterile needle which provided it with the necessary stimulus to unde rgo all the
cytoplasmic and biochemical rearrangements associated with fertilization, including the
completion of second meiotic division at the animal pole of the cell. Puncturing the
oocyte at this point let the spindle and chromosomes flow out of the ce ll (enucleation).
The nucleus from a donor cell was then removed and by using a micropipette was
inserted into this activated, enucleated cell. Briggs and King demonstrated that blastula
(early-stage embryo) nuclei when transferred into activated enucleate d oocytes could
direct the development of a completely formed tadpole. They also found that while the
nuclei at the blastula stage were totipotent, there was a dramatic decrease in the nuclear
potency of cells at later stages. For instance, when nuclei fro m somatic cells of the tail
bud region of tadpoles were used in a similar experiment, normal development did not
occur. Thus, it was concluded that nucleus from developing embryonic cells appeared to
lose the ability to direct development as they underwent determination and
differentiation. Further work continued using nuclear transplantation studies and in
1962 John Gurdon a PhD student in Cambridge University showed that if the nucleus of
a fertilized frog egg was replaced with a nucleus from the cell of the tadpole intestine,
the egg could develop into a new frog. Though the success rate was low, it proved that
the nucleus of a mature cell still contained the genetic information needed to build all
cell types. This was a major landmark in animal developme nt though the
acknowledgment of the importance of his work came much later. It was 40 years after

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his first experiments with nuclear transformation that the Nobel Prize was awarded in
2012 to Gurdon along with Shinya Yamanake, whose lab induced pluripotent cells.
Nuclear transplantation techniques were used by experimental embryologists to create
clones of several species over the years. In 1997 Ian Wilmut and his colleagues
demonstrated that somatic cells still had the potency to produce all the cells o f the
body. The result was Dolly, a sheep that was cloned by using the nucleus taken from the
mammary gland of an adult sheep and transplanting it into the egg taken from another
strain of sheep. This was done in a culture and later the eggs containing the
transplanted nucleus were implanted into the uteri of pregnant sheep. Of the 434 sheep
oocytes only one survived to become Dolly. DNA analysis however, confirmed that
Dolly’s cells had indeed been derived from the donor nucleus. This was the final proof
of genomic equivalence of somatic cells and that the genome is conserved during
differentiation of cells. Since the first cloning experiments, cloning of adult mammals
has been done in mice, rats, guinea pigs, rabbits, dog, cat, cows and horses.
Differential Gene Expression
Genomic equivalence established the general principle that somatic cell nucleus
of an organism contains the complete genome established at the time of fertilization and
there is no change in the genetic content after differentiation of cells. If all cells in the
developing embryo have the same genetic content, differences between cells must be
due to different gene activity in each cell type. This means that only a small portion of
the genome is expressed or “turned on” in each cell. As a result, only those RNAs are
synthesized in the cell that will translate (make) only those proteins that will provide
structural information to these cells to help them differentiate and ultimately develop
into very different cell types. It is important to understand how var ious signals cause
cells to express different portions of their genetic information. Nuclear transplantation
and cell fusion experiments reveal that gene activity is controlled by the cytoplasmic
environment. New cytoplasmic signals can activate previously silent genes and silence
previously active genes. Such genetic control continues throughout embryonic
development and results in the generation of a variety of cell types with different gene
activities. As the zygote cleaves, its cytoplasmic contents cont ributed by the egg do not
pass uniformly into different blastomeres. The descendent cells at cleavage, therefore,
have different cytoplasmic environments. This initiates a pattern of embryonic cells
with a distinct programme for gene expression. Later in e mbryonic development,
interactions between blastomeres release new signals, which then determine additional
group of cells to activate new sets of genes.
The first evidence for differential gene expression came from the study of
polytene chromosomes of Drosophila larva. In polytene chromosomes DNA duplication
occurs in many rounds without any division and separation of the cell. The cell size
increases and so as a result, the many stranded DNA is easy to observe. It was seen that

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the banding pattern of the chromosomes was identical throughout the various cells of
the larva. No loss or addition of any chromosome portion was seen in different cell
types in the larva. However, different parts of the chromosome were ‘puffed up’ in
different cell types suggesting that different RNA was being synthesized in different
cells and while one part was puffed up in one cell type the other genes were silent and
so not puffed up in that cell
By the late 1980s it was understood that gene expression could be regulated basic ally at
4 steps and this could explain the mechanism of differential gene expression.
i) At the stage of gene transcription so that only those genes that are required for that
particular cell type are expressed in the form of nuclear RNA. Transcription is the first
step of gene expression and its control involves general and tissue specific
transcriptional regulators.
ii) At the nuclear RNA processing stage, by regulating which part of the RNA or which
of the transcribed RNAs are allowed to leave the nucl eus.
iii) At the RNA translation stages by regulating which of the mRNAs are to be
translated into proteins.
iv) At the protein modification stage, so that only those proteins are modified that can
provide the structure and function to the specific cell type.
A gene is a distinct sequence of the DNA molecule that has the information to make a
polypeptide or a nucleotide. Figure 3.7 shows the gene responsible for making the β -
globin in the haemoglobin molecule. The β-globin is made up of different components.
Only the parts labelled exons will provide the information for the appropriate amino
acid sequences for translating the appropriate protein and not the parts labelled intron.
On this portion of DNA, the transcription will start at the transcription in itiation site
and end at transcription termination site. Thissegment will form the HnRNA
(heterogeneous n RNA) and will undergo processing in which the transcribed introns
will be spliced or removed. This would the result in putting together the consecutiv e
exons and forming the mRNA. In further modification, 7 -methylguanosine (G) cap and
a poly (A) tail will be added to the 5-prime end and 3- prime end of this mRNA
respectively. This modified mRNA will then leave the nucleus to move into the
cytoplasm where it will be translated into a protein of interest. Look at the gene again
in the figure 3.7. You will see that the DNA has a portion marked promoter which is not
transcribed. The promoter is very important as it is at this location where the proteins
and factors bind which can thus regulate or enable the process of transcription.
Furthermore, in the upstream (anterior end) region or in some cases even in downstream
(posterior end) region and also in the introns, there will be sequences that will be able
to influence the promoter to initiate transcription. These sequences are regulatory
factors also known as enhancers (that stimulate), repressors or silencers (that inhibit
transcription).Let us now see how regulation of gene expression can occur. Earlier in

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the section we had said that gene expression can be regulated at all stages from
transcription to the final making of the gene product, namely protein. We will discuss
the regulation at the transcription level in a little more detail so that you can unders tand
this important mechanism. If a gene is to be regulated then first it should be accessible
to a variety of factors that can bind to the DNA and regulate its expression.
1) The first step would be the loosening of the chromosome so that it can change from
hetrochromatin state (when the DNA is packaged and coiled around the histones) to the
euchromantin state (when the uncoiling of the DNA occurs) so that the gene becomes
accessible to a variety of factors. Therefore, regulating the chromatin to uncoil or coil
will influence where and when in the genome certain genes can be expressed. For this
process small organic molecules can be added or removed from the histones around
which the DNA remains coiled. DNA remains coiled if the histones have methyl group s
attached to their tails and uncoiling occurs when the methyl groups are replaced by
acetyl groups on the histone tails. It is a general rule that acetylation will give access to
the promoter region and initiate active transcription, while methylation wil l repress
transcription. Experiments have shown that this is one way of regulating gene
expression.
2) Another way of regulation is by transcription factors that bind to the promoter region
and can turn genes on or off. If even one transcription factor is changed in the embryo
it can have dramatic effects. For example, the loss of a gene ultrabiothorax c an have
drastic effects in insects. This gene is a homeobox gene which contains a transcription
factor, whose absence changes the segmentation pattern in the fly. As a result, the fly
forms two pairs of wings instead of the usual one pair due to loss of a gene
ultrabiothorax. There are different transcription factors which bind to their affiliated
DNA binding domains and these are very specific. Let us take the example of an
enhancer region on a gene. A transcription factor specific to that enhancer forms a
complex with it and can change the shape of DNA so that the promoter and
transcription factor- enhancer complex come near the promoter. This causes a very
important enzyme DNA polymerase to bind to the promoter which causes the
transcription of a particular RNA. If transcription factor binds to the repressor region it
will stop or slow down the gene from expressing. For example, the human foetal
embryo liver cells synthesize serum albumin but only after a certain stage of
development. Till then the gene that encodes for serum albumin is silent or repressed as
a transcription factor is bound to the repressor domain. Thus, again it is the specific
combination of the transcription factor with the enhancer or repressor that will regulate
the rate of transcription in specific cells and cause differential gene expression. Often
there is a cascade of reactions in which the gene for making a transcription factor is
regulated by another transcription factor.

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Therefore, it is important for you to understand that the re are a variety of ways to
regulate the gene and the protein it forms, which in turn influence the differentiation of
the incredible varieties of cells seen. Scientists have still a lot to learn and discover
about these mechanisms.

Fig. 4.3.5: Beta-globin synthesis for hemoglobin - Inactive until modified and
combined with alpha globin to form complete hemoglobin molecule .
CELL DETERMINATION AND DIFFERENTIATION
In the earlier section, we explored how fate maps reveal the areas of the embryo
that contribute to various tissues and organs in the adult, while cell fate outlines what a
cell will become during normal development. To trace a cell's destiny, labeling
techniques are employed, observing the structure it integrates into. The developmental
potential, or potency, of a cell refers to the range of cell types it can give rise to. In the
initial stages, the zygote and early blastomere are totipotent, capable of developing into
a complete organism. However, totipotency diminishes after the first few divisions in
the blastula.As development progresses, individual cells' developmental potential
decreases until their fate is determined. This commitment means each cell is dedicated
to forming a specific structure, following a particular developmental path. Co nversely,
differentiation is the process where cells cease dividing and acquire the distinctive
structure and functions of a specific cell type. It represents the final stage where an
undifferentiated cell undergoes a series of events to become a specializ ed cell, such as a
muscle, nerve, or skin cell.
CELL DETERMINATION
The process of commitment takes place in two steps:

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a) Specification: The fate of the cell is known to have been specified whenit is capable
of differentiating autonomously even if it is placed in an environment that is neutral
like a culture medium in a petri dish (Fig.3.4 a). However, at this stage the commitment
of the cell is flexible and it can be influenced by its environment to become another
type of cell rather than what it was specified or fated to be (see Fig.3.4 b).
b) Determination: this step is after specification of the cell to form a particular type of
cell. In the stage of determination of the cell the fate of the cell becomes inflexibly or
irreversibly specified or determined to form a particular type of cell. As a result, the
cell will still autonomously differentiate into its original specified fate and its
differentiation into aparticular type of cell will be independent of its environment or its
position in the embryo it (see Fig.4. 3.4 c).

Fig. 4.3.6: Isolated blastomeres from blastula, one specified for muscle and the other
for neurons. a) Muscle-specified differentiates into muscle cells, neuron -specified into
neurons; b) Muscle-specified in neuronal tissue cluster becomes neural cells; c)
Determined muscle blastomere, even in neuronal tissue, differentiates into muscle cells.
Induction:

Induction in developmental biology refers to the process by which one group of cells or
molecules influences the fate or behavior of neighboring cells or molecules, leading to
changes in their development or differentiation. It plays a crucial role in various stages
of embryonic development, including pattern formation, cell differentiation, and tissue
formation.
There are two main types of induction:
Paracrine Induction: This occurs when signaling molecules, such as growth factors or
hormones, released by one group of cells act locally on nearby cells to induce specific
responses. Paracrine induction is essential for the establishmen t of gradients of
signaling molecules, which in turn determine cell fate along the embryonic axes.
Neighboring Cell Induction: Also known as juxtacrine induction, this type of
induction involves direct physical contact between neighboring cells, where cell surface
molecules or ligands on one cell interact with receptors on another cell, triggering

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