Divine Grace E.

Valenzuela MABioEd

MABioEd215: Developmental Biology

Midterm Examination

1. What is the central position and impact to society of developmental biology? How would
developmental biology shape the future of research and science education?
Developmental Biology is the study of the processes by which organs grow and
develop. Modern developmental biology studies the genetic control of cell growth,
differentiation and morphogenesis, which is the process that gives rise to tissues, organs
and anatomy, but also regeneration and aging. Also it is the study of the process by which
animals and plants grow and develop, and is synonymous with ontogeny and the causal
analysis of the cellular mechanisms that drive processes of growth, pattern formation and
morphogenesis. There are many important arguable biological discipline linked to
developmental biology that serve a great impact to society of this branch of biology hence
here are some instances (1) developmental biology addresses the key challenge of
population health especially when it comes to sustaining food resources which is a major
global challenge and developmental biology can provide strategies to improving crop and
cultivation (2) developmental biology continues deliver the most effective explanations for
diseases or medically relevant processes including infertility, neonatal death, birth defects
(deformation, body growth abnormalities, developmental brain disorders), cancer, wound
healing and tissue regeneration including stem cell and it is done through medical
researches using animal as a model.

2. Fertilization is the process whereby two sex cells (gametes) fuse together to create new
individual with genetic potentials derived from both parents. Although that this varies from
species to another, discuss four major events in fertilization, the gametes involved
considering its anatomy and physiology and the prevention of polyspermy.
Fertilization is the process whereby the sperm and the egg—collectively called the
gametes—fuse together to begin the creation of a new individual whose genome is
derived from both parents. Fertilization accomplishes two separate ends: sex (the
combining of genes derived from two parents) and reproduction (the generation of a new
organism). Thus, the first function of fertilization is to transmit genes from parent to
offspring, and the second is to initiate in die egg cytoplasm those reactions that permit
development to proceed. Although the details of fertilization vary from species to species,
conception generally consists of four major events:
1. Contact and recognition between sperm and egg. In most cases, this ensures that the
sperm and egg are of the same species.
2. Regulation of sperm entry into the egg. Only one sperm nucleus can ultimately unite
with the egg nucleus. This is usually accomplished by allowing only one sperm to enter
the egg and actively inhibiting any others from entering.
 3. Fusion of the genetic material of sperm and egg.
4. Activation of egg metabolism to start development.

Structure of the Gametes

Sperm

Sperms are highly motile cells consisting of nucleus, mitochondria to provide

energy source and a flagellum for movement. The anterior end of the sperm is highly
specialized which aids in penetration of the egg. Sperms are typically designed to activate
the egg and to deliver their nuclei into the egg cytoplasm.

Sperm Formation

Sperm Axoneme

The Egg

Eggs are non-motile, surrounded by protective egg coverings. These serve to

recognize the sperm specifically and prevent fertilization by more than one sperm
(polyspermy). The mammalian egg has zona pellucida layer around the plasma
membrane beneath which cortical granules are present. The zona pellucida layer makes
the egg impenetrable to more than one sperm.

All material necessary for the beginning of growth and development must be stored
in the egg, or ovum. Where-as the sperm eliminates most of its cytoplasm as it matures,
the developing egg (called the oocyte before it reaches the stage of meiosis at which it is
fertilized") not only conserves the material it has, but actively accumulates more. The
cytoplasmic trove includes the following: (1) Nutritive proteins. It will be a long time
before the embryo is able to feed itself or even obtain food from its mother, so the early
embryonic cells need a supply of energy and amino acids. In many species, this is
accomplished by accumulating yolk proteins in the egg. Many of these yolk proteins are
made in other organs (e.g. liver, fat bodies) and travel through the maternal blood to the
oocyte. (2) Ribosomes and tRNA. The early embryo needs to make many of its own
structural proteins and enzymes, and in some species there is a burst of protein synthesis
soon after fertilization. Protein synthesis is accomplished by ribosomes and tRNA that
exist in the egg. (3) Messenger RNAs. The oocyte not only accumulates proiteins it also
accumulates mRNAs that encode proteins for the early stages of development. (4)
Morphogenic factors. Molecules that direct the differentiation of cells into certain cell
types are present in the egg. These include transcription factors and paracrine factors. In
many species, they are localized in different regions of the egg and become segregated
into different cells during cleavage. (5) Protective chemicals. The embryo cannot run
away from predators or move to a safer environment, so it must come equipped to deal
with threats. Many eggs contain ultraviolet filters and DNA repair enzymes that protect
them from sunlight. Some eggs contain molecules that potential predators find distasteful,
and the yolk of bird eggs even contains antibodies.

Enclosing the cytoplasm is the egg plasma membrane. This membrane must
regulate the flow of certain ions during fertilization and must be capable of fusing with the
sperm plasma membrane. Outside the plasma membrane is the vitelline envelope, which
forms a fibrous mat around the egg. This envelope contains at least eight different
glycoproteins and is often involved in sperm-egg recognition (Correia and Carroll 1997).
It is supplemented by extensions of membrane glycoproteins from the plasma membrane
and by proteinaceous vitelline posts that adhere the vitelline envelope to the membrane
(Mozingo and Chandler 1991). The v itelline envelope is essential for the species-specific
binding of sperm. In mammals, the vitelline envelop e is a separate and thick extracellular
matrix called the zona pellucida. The mammalian egg is also surrounded by a layer of
cells called the cumulus, which is made up of the ovarian follicular cells that were
nurturing the egg at the time of its release from the ovary. Mammalian sperm have to get
past these cells to fertilize the egg. The innermost layer of cumulus cells, immediately
adjacent to the zona pellucida, is called the corona radiata. Lying immediately beneath
the plasma membrane of the egg is a thin shell (about 5 μm) of gel-like cytoplasm called
the cortex. The cytoplasm in this region is stiffer than the internal cytoplasm and contains
high concentrations of globular actin molecules. During fertilization, these actin molecules
polymerize to form long cables of actin known as microfilaments. Microfilaments are
necessary for cell division, and they also are used to extend the egg surface into small
projections called microvilli, which may aid sperm entry into the cell. Also, within the
cortex are the cortical granules.

Recognition of Egg and Sperm

The interaction of sperm and egg generally proceeds according to five basic steps
which are the following (1) The chemoattraction of the sperm to the egg by soluble
molecules secreted by the egg. (2) The exocytosis of the acrosomal vesicle to release its
enzymes. (3) The binding of the sperm to the extracellular envelope (vitelline layer or zona
pellucida) of the egg. (4) The passing of the sperm through this extracellular envelope (5)
Fusion of egg and sperm cell plasma membranes. Sometimes steps 2 and 3 are reversed
(as in mammalian fertilization) and the sperm binds to the egg before releasing the
contents of the acrosome. After these five steps are accomplished, the haploid sperm and
egg nuclei can meet, and the reactions that initiate development can begin

Sperm attraction: In many animals, sperms are attracted towards eggs of their species
by “chemotaxis” i.e. following a gradient of a chemical secreted by the egg. Chemotaxis
has been demonstrated in cnidarians, molluscs, echinoderm and urochordates (Miller,
1985; Yoshida et al, 1993). Similarly, in the egg jelly of the sea urchin, chemotactic factors
are present for sperm attraction.
Gamete binding and recognition in mammals:
 In mammals fertilization is internal. The reproductive tract plays a very active role
in fertilization. The differentiated sperms are unable to undergo the acrosome reaction
without residing for some time in the female reproductive tract where they undergo
physiological changes. The change in the mammalian spermatozoan, which makes it
capable of fertilizing the egg, is called capacitation.
The mammalian egg is surrounded by extracellular envelope called zona pellucida.
Around zona pellucida is a layer of cumulus cells (corona radiate) embedded in a
cementing substance, hyaluronic acid. Hyaluronidase activity on the surface of the sperm
head helps it to penetrate this layer. Next, sperm must bind to zona pellucida before they
make contact with the surface of egg itself. The zona pellucida in mammals plays a role
analogous to that of viteline envelope in lower vertebrates and invertebrates. The zona
pellucida is a glycoprotein matrix synthesized and secreted by the growing oocyte. It plays
two important roles in fertilization. It binds the sperm and initiates acrosome reaction.
Binding of the spermtozona triggers the acrosome reaction, which allows the
sperm to penetrate the zona. Acrosome reaction in mammals involves the fusion of the
outer membrane of the acrosome with the sperm plasma membrane. After the fusion, the
acrosomal membrane vesiculates which results in the release of acrosomal contents.
Subsequently, the outer portion of the acrosomal membrane disappears and only the inner
portion adjacent to the nucleus remains intact. When the acrosomal contents are
exocytosed, several enzymes are released. These enzymes allow the sperm to approach
the egg plasma membrane. The mammalian acrosome reaction differs from that of sea
urchin in that no acrosomal process is formed. A sperm protein fertilin, is thought to be
involved in mediating fusion of sperm egg membrane.

The prevention of polyspermy

The entry of the sperm into the egg activates the egg. Although many sperm attach
to the egg surface, it is important that only one sperm enters the egg (monospermy). Entry
of more than one sperm (polyspermy) may result in several abnormalities such as
polyploidy, abnormal mechanism of chromosomal separation during cell division and
ultimate death of the embryo. Organisms have evolved ways to prevent the union of more
than two haploid nuclei.
In fishes the sperm can enter into the egg only through the narrow opening, the
micropyle, the rest of the egg being covered by impermeable chorion.
In sea urchin and mammals, there is restriction on the number of sperm that are
able to penetrate the extra cellular coats and fuse with the egg. In mammals, the sperm
has to migrate the long female reproductive tract to reach the egg and further, structural
changes in zona pellucida block polyspermy. The most common way to prevent
polyspermy is to prevent the entry of more than one sperm into the egg. The polyspermy
is blocked in many animals as soon as the first sperm fuses with the egg plasma
membrane.

The activation of egg metabolism:

 After the sperm penetrates the egg a series of diverse cytoplasmic reactions are
initiated. The response of the egg to the sperm can be divided into “early” responses,
which occur within seconds of the cortical reaction and “late” responses which take place
several minutes after fertilization begins.

Fusion of genetic material

In mammals the entry of sperm is tangential to the eggs surface. Once inside the
egg, the sperm nucleus becomes male pronucleus. The sperm centrosome produces
asters and contacts the female pronucleus. Male & female pronuclei migrate towards each
other, become apposed but do not fuse. They remain adjacent to each other; their nuclear
envelopes break down but instead of forming a zygote nucleus the chromatin condenses
into chromosomes orienting them on a common mitotic spindle. Thus, only after
completion of the first division of fertilized egg, the paternal and maternal chromosomes
become enclosed by a common nuclear membrane to form the nuclei of two blastomeres.

3. Given the emphasis on the anatomy of gene, discuss the physiology of exons and introns,
transcription factors, promoters and enhancers and the elements of gene expression.
Transcription, the synthesis of rRNA, tRNA, and mRNA using the information
from DNA. The process is similar to replication. A portion of the DNA unwinds to form a
transcription bubble. One of the two DNA strands will act as the template strand for RNA
synthesis. Eukaryotic genes are discontinuous; not all of the DNA sequence codes for
protein. Some segments are between coding sequences. The sequences that do not
encode any amino acid sequences for the protein are called intervening sequences or
introns. The protein coding sequences are called the exons. The ptRNA, the first DNA-
directed mRNA product contains both the introns and the exons. But the presence of
introns in the mRNA would make it impossible for the process of translation to synthesize
the correct protein. RNA splicing is done to remove these portions.

Transcription factors are proteins involved in the process of converting, or

transcribing, DNA into RNA. Transcription factors include a wide number of proteins,
excluding RNA polymerase, that initiate and regulate the transcription of genes. One
distinct feature of transcription factors is that they have DNA-binding domains that give
 them the ability to bind to specific sequences of DNA called enhancer or promoter
sequences. Some transcription factors bind to a DNA promoter sequence near the
transcription start site and help form the transcription initiation complex. Other transcription
factors bind to regulatory sequences, such as enhancer sequences, and can either
stimulate or repress transcription of the related gene. These regulatory sequences can be
thousands of base pairs upstream or downstream from the gene being transcribed.
Regulation of transcription is the most common form of gene control. The action of
transcription factors allows for unique expression of each gene in different cell types and
during development. Transcription factors can activate or repress the transcription of a
gene, which is generally a key determinant in whether the gene functions at a given time.

4. In the discussion of animal development, the specification of early embryonic cells by their
acquisition of different cytoplasmic determinants are being stored in the oocyte. The cell
membranes establish the region of cytoplasm incorporated into each new blastomere, and
it is thought that the morphogenetic determinants then direct differential gene expression
in these blastomeres. Given these emphases, this is different from Drosophila
development, discuss further and explain the mechanism of Drosophila development,
including polarity and the effects of various genes.

The Drosophila life cycle consists of a number of stages: embryogenesis, three

larval stages, a pupal stage, and (finally) the adult stage.

Cleavage

The eggs you collect will be in various stages of development. It is difficult to see
what is going on inside the egg both because of the chorion (if you haven’t removed it)
and because the egg is very yolky. It is a centrolecithal egg, meaning that the yolk is
concentrated centrally and the cytoplasm is pushed to the periphery. Cleavage is unusual
in that nuclear division (karyokinesis) occurs many times before the cytoplasm cleaves
(cytokinesis). The nuclei of early cleavage are centrally located through the first seven
divisions, after which they start migrating to the periphery. By the time there are about
5000 nuclei all lined up in the peripheral cytoplasm, cell membranes are laid down
between them, making a peripheral layer of separate cells, and the embryo goes from
being a syncytial blastoderm to a cellular blastoderm. This pattern of cleavage is called
superficial or peripheral cleavage. Cellularization of the blastoderm does not occur
simultaneously around the egg. The cells that form first are at the posterior end. They are
relatively large and are called the pole cells. The pole cells form the primordial germ cells,
which give rise to the gamete.
 Gastrulation

Following cleavage, gastrulation proceeds primarily by the infolding of a midventral

band of cells. First a ventral furrow appears as the mesoderm folds inward. At the anterior
and posterior ends of this furrow, the endoderm invaginates forming the anterior and
posterior midgut. Later the ectoderm will also invaginate to form the anterior stomodeum
(foregut) and posterior proctodeum (hindgut). In addition to these ventral infoldings, there
is also a lateral infolding toward the anterior end which extends around the circumference
of the embryo. This is the cephalic furrow and roughly delineates the boundary of the
future head. Gastrulation creates a multilayered band of germ layers on the ventral side
of the egg that curves around the egg’s posterior tip. This band is called the germ band. It
elongates along the dorsal side of the egg so that eventually, like an acrobat with her back
arched and her legs gracefully curved back to touch her head, the embryo’s posterior end
meets its head end (Figures 8.2G, H). The germ band then shortens and thickens, bringing
the posterior end of the embryo back toward the posterior pole of the egg. As the germ
band shortens, definitive segmental boundaries appear, marking off head regions
(mandibular, maxillary, labial), thoracic segments (t1–t3), and abdominal segments (a1–
a10). (In the larva you will find only eight abdominal segments. Abdominal segments 9
and 10 have formed the telson of the larva, a tail-like structure.). During gastrulation, a
peculiar thing happens. The developing head disappears from view—it turns inward, or
involutes. Meanwhile the thoracic segments expand forward, overgrowing the region that
used to be “head.” Only a tiny external head will remain.

Later development

By 16 hours of development, muscular movement will be apparent (Figure 8.2L). Just

before the embryo hatches as the first instar larva at 22–24 hours, you will be able to see air-
filled tracheae and other internal organs.
 The maternal-effect genes, including bicoid and nanos, are required during
oogenesis. The transcripts or protein products of these genes are found in the egg at
fertilization, and form morphogen gradients. The maternal-effect genes encode transcription
factors that regulate the expression of the gap genes. The gap genes roughly subdivide the
embryo along the anterior/posterior axis. The gap genes encode transcription factors that
regulate the expression of the pair-rule genes. The pair-rule genes divide the embryo into
pairs of segments. The pair-rule genes encode transcription factors that regulate the
expression of the segment polarity genes. The segment polarity genes set the
anterior/posterior axis of each segment. The gap genes, pair-rule genes, and segment
polarity genes are together called the segmentation genes, because they are involved in
segment patterning

5. Vertebrate axes do not form from localized determinants in the various blastomeres,
considering this factor, discuss and explain the axis formation in amphibians and the
actions of various factors and genes in these progressive developmental changes.

Vertebrate axes do not form from localized determinants in the various

blastomeres, as in Drosophila. Rather, they arise progressively through a sequence of
interactions between neighboring cells. Amphibian axis formation is an example of
regulative development.

Cleavage in Amphibians

In eggs that contain moderate to little yolk, cleavage occurs throughout the whole
egg, a pattern called holoblastic cleavage. This pattern of cleavage is characteristic of
invertebrates such as mollusks, annelids, echinoderms, and tunicates, and also of
amphibians and mammals. Amphibian eggs contain much more cytoplasmic yolk in the
vegetal hemisphere than in the animal hemisphere. Because yolk-rich regions divide much
more slowly than areas with little yolk, horizontal cleavage furrows are displaced toward
the animal pole.
 An amphibian embryo containing 16 to 64 cells is commonly called a morula
(plural: morulae; from the Latin, "mulberry," whose shape it vaguely resembles). At the
128-cell stage, the blastocoel becomes apparent, and the embryo is considered a
blastula.

While these cells are dividing, numerous cell adhesion molecules keep the
blastomeres together. One of the most important of these molecules is EP-cadherin. The
mRNA for this protein is supplied in the oocyte cytoplasm. If this message is destroyed
(by injecting antisense oligonucleotides complementary to this mRNA into the oocyte), the
EP-cadherin is not made, and the adhesion between the blastomeres is dramatically
reduced (Heasman et al. 1994a,b), resulting in the obliteration of the blastocoel.

Gastrulation in Amphibians

The blastula of an amphibian has an asymmetrical yolk distribution, and the yolk-
laden cells of the vegetal pole are less numerous but much larger than the yolk-free cells
of the animal pole. Consequently, gastrulation is more complex than it is in sea urchins.
In frogs, a layer of surface cells first invaginates to form a small, crescent-shaped slit,
which initiates formation of the blastopore. Next, cells from the animal pole involute over
the dorsal lip of the blastopore, which forms at the same location as the gray crescent of
the fertilized egg. The involuting cell layer eventually presses against the inner surface of
the opposite side of the embryo, eliminating the blastocoel and producing an archenteron
with a blastopore. In this case, however, the blastopore is filled with yolk-rich cells, forming
the yolk plug. The outer layer of cells resulting from these movements is the ectoderm,
and the inner layer is the endoderm. Other cells that involute over the dorsal lip and ventral
lip (the two lips of the blastopore that are separated by the yolk plug) migrate between the
ectoderm and endoderm to form the third germ layer—the mesoderm.

6. Describe and explain the differences and similarities of vertebrate and inverterbrate
development considering the genes and factors affecting the onset of these animals’
development.
Early Development in Invertebrates
During cleavage, most cells do not grow. Rather, the volume of the oocyte is
cleaved into numerous cells. The major exceptions to this rule are mammals. The
blastomere cell cycle is governed by the synthesis and degradation of cyclin. Cyclin
synthesis promotes the formation of MPF, and MPF promotes mitosis. Degradation of
cyclin brings the cell back to the S phase. The G phases are added at the midblastula
transition. "Blast" vocabulary: A blastomere is a cell derived from cleavage in an early
embryo. A blastula is an embryonic structure composed of blastomeres. The cavity in the
blastula is the blastocoel. If the blastula lacks a blastocoel, it is a stereo blastula. A
mammalian blastula is called a blastocyst, and the invagination where gastrulation begins
is the blastopore. The movements of gastrulation include invagination, involution,
ingression, delamination, and epiboly. Three axes are the foundations of the body: the
anterior-posterior axis (head to tail or mouth to anus), the dorsal-ventral axis (back to
belly), and the right-left axis (between the two lateral sides of the body). In all four
invertebrates described here, cleavage is holoblastic. In the sea urchin, cleavage is radial;
in the snail, spiral; in the tunicate, bilateral; and in the nematode, rotational.

Early Development in Vertebrates

Fishes, reptiles, and birds undergo discoidal meroblastic cleavage, wherein the
early cell divisions do not cut through the yolk of the egg. These cells form a blastoderm.
In fishes, the deep cells form between the yolk syncytial layer and the enveloping layer.
These cells migrate over the top of the yolk, forming the hypoblast and epiblast layers. On
the future dorsal side, these layers intercalate to form the embryonic shield, a structure
homologous to the amphibian organizer. Transplantation of the embryonic shield into the
ventral side of another embryo will cause the formation of a second embryonic axis.
Mammals undergo holoblastic rotational cleavage, characterized by a slow rate of division,
a unique cleavage orientation, lack of divisional synchrony, and the formation of a
blastocyst. The blastocyst forms after the blastomeres undergo compaction. It contains
outer cells the trophoblast cells that become the chorion, and an inner cell mass that
becomes the amnion and the embryo. The chorion forms the fetal portion of the placenta,
which functions to provide oxygen and nutrition to the embryo, to provide hormones for
the maintenance of pregnancy, and to provid barriers to the mother's immune system.
Mammalian gastrulation is not unlike that of birds. There appear to be two signaling
centers one in the node and one in the anterior visceral endoderm. The latter is critical for
generating the forebrain, while the former is critical in inducing the axial structures caudally
from the midbrain. Hox genes pattern the anterior-posterior axis and help to specify
positions along that axis. If Hox genes are knocked out, segment-specific malformations
can arise. Similarly, causing th ectopic expression of Hox genes can alter the body axis.
 7. Construct a session plan for Developmental Biology indicating Learning Outcomes,
Performance Indicators, Learning Tasks (Subject Matter, Materials, References,
Concepts, Skills). Procedure and Assessment (use the topics to you individually to
construct your session plan). This must include the use of various teaching approaches
on how to teach the subject matter. (separate sheet)

Learning Tasks Learning Outcomes Performance Indicators

A. Subject Matter The learner is able to… At the end of the lesson, the
Development and learners will be able to:
Disease
B. Materials 1. Identify various 1. Explain the various
Laptop, Projector associated diseases associated diseases
whiteboard, whiteboard during development. during development
marker
C. References 2. Identify the four 2. Describe the four
Gilbert, Scott F. (2003). principles of how principles of how
Developmental Biology teratogens affect teratogens affect
7th edition. development. development and name
15.2 Cunningham, some teratogens in the
W.P. el.al (2009). 3. Design a poster showing environment.
Principles of the harmful effects of
Environmental Science teratogens in 3. Create an informational
6th ed. McGraw-Hill development. poster about the harmful
Companies Inc. New effects of teratogens in
York. development.
C. Concepts
 Abnormalities due to
exogenous agents
(certain chemicals or
viruses, radiation, or
hyperthermia) are called
disruptions.
 The agents responsible
for these disruptions are
called teratogens
(Greek, "monster-
formers")
 The study of how
environmental agents
disrupt normal
development is called
teratology