COMPARATIVE VERTEBRATE MORPHOGENESIS ZOO

4690
FLORIDA ATLANTIC UNIVERSITY @ DAVIE BROWARD
CAMPUS
Professor: James Kumi-Diaka (BSc. DVM, MSc, PhD. Vee-dip)
Office: Science & Education Bldg. Room 278 Phone: (954)236-1135

Course Objective:
5. To expose students to the concept and principles of
developmental biology (embryology and gross anatomy) with
regard to structural and functional physiological development in
vertebrates.
Course Outline:
7. Introduction: historical perspective; phylum chordata; its
characteristic features; concepts of homology and analogy.
8. Vertebrate phylogeny: classification.
9. Gametogenesis: definitions; nuclear and cytoplasmic; cell
divisions; purpose of.
10. Fertilization: What is it?; purpose of; the process of; end-product
of; parthenogenesis.
11. Early development: cleavage/segmentation holoblastic and
meroblastic cleavages; formation of morula and blastula.
12. Gastrulation: the formation of gastrula; the process involved;
formation of primary germ layers; extraembryonic membranes
and their role in development.
 Course Outline cont’d
1. Induction: principle of; physiological significance; classification of.
2. Placentation: amniotes and anamniotes; classification; functions of.
3. The integument: components of: structure and derivatives of.
4. Skeletal system I: components of; skull development; morphology.
5. Skeletal system II: origin and developmental anatomy of vertebrae.
6. Skeletal system III: appendicular skeleton, origin, development and
morphology.
7. Musculature I: muscle origin; fate of myotomes: classification;
histoanatomy and topography.
8. Muscular system II: development of brachiomeric & appendicular
muscles.
9. Development of coelomic cavities, and mesenteries.
10. Respiratory system: evolution, morphogenesis. and derivatives of;
classification.
11. Digestive system: evolution and morphogenesis, derivatives of,
classification of.
12. Urogenital system I: kidney structure; morphogenesis; classification;
primordial gonads.
13. Urogenital system II: reproductive system; embryogenesis and
morphoanatomy.
14. Cardiovascular system I: circulatory system; morphogenesis of
extraembryonic circulation.
15. Cardiovascular system II: aortic arches and their derivatives.
16. Cardiovascular system III: the heart - its embryonic origin and
 Course Outline cont’d
1. Nervous system I: morphogenesis and histoanatomy of
neural tube (brain and spinal cord).
2. Nervous system II: development of autonomic nervous
system, spinal nerves, spinal cord.
3. Nervous system llI: cranial nerves, special senses, olfactory,
optic, otic, etc.

TEXT BOOK(S): Any of the following text books will be OK

1. Analysis of Biological Development –Klaus Kalthoff, 2nd
Ed.
2. Patten’s Foundations of Embryology 6th Edit, Bruce M
Carlson
3. Principles of Development 2nd Edit, Lewis Wolpert
 EXAMINATIONS;
Exam I = 02/04/08 100%
Exam II = 03/12/08 100%
Exam III = 04/21/08 100%

 EVALUATIONS:
 A >90%; B = 80-89%; C = 70-79%: D = 60 – 69%: F = < 60

PLEASE NOTE:
 i) All make-ups exams will draw 15% penalty
 ii) Exam results will NOT be given over the phone
(do not).
 III) THERE ARE NO EXTRA-CREDITS
 IV) NO CELL PHONES ALLOWED DURING EXAMS
–
25%
PENALTY
 EMBRYOLOGY
 Zygote - all higher animals start their lives from a single cell
zygote - dual origin from two gametes – spermatozoon + ovum

 Ontogeny - The time of fertilization represents the starting point in

the
life history
- Ontogeny refers to the individual’s entire life span.

 German biologist August Weismann (1834-1914) made the

important distinction between the soma (body) and the germ-cell
line (gametes).
 - thought that the germ-cell line was all-important for perpetuation
of the
species, and that the soma was primarily a vehicle for protecting
and
perpetuating the germ plasm.
 This viewpoint provides a convenient framework for looking at the
perpetuation of life (Fig. 1-2).

 Once an individual has passed the reproductive

years, the remainder of its ontogeny does not
provide direct physical input into the generative
 Historical perspective

 The origin of the embryo: Aristotle’ Theories

i) Preformationist concept
- everything in the embryo was preformed from
the
very beginning and dimply got bigger during

development
- the sperm contained the embryo (homunculus
– tiny
human in the head of the sperm)

ii) Epigenesis (means ‘upon formation’)

- new structures arose progressively during
development
 FUNDAMENTAL PROCESSES AND
CONCEPTS IN DEVELOPMENT
Cell Division and the Cell Cycle
 Is the fundamental properties of living systems, of vital importance

 Cell division in vertebrates takes two forms—mitosis and meiosis.

 Mitosis (for review, see Fig. 1-14) is the standard form of cell division in
somatic cells and it results in two genetically equal daughter cells.

 Meiosis (Fig. 3-7) is confined to certain stages of development of

gametes
 Mitosis is one component of the cell cycle.

 The life history of a cell can be divided into four periods (Fig. 1-15).
Immediately after mitosis and the separation of the dividing cell into
daughter cells, the G1 (gap I) period, often called the interphase
commences. Its length is extremely variable. In rapidly cleaving
embryos just after fertilization, the G1 phase is very short and
sometimes may not even exist.
 At the other extreme, the G1 phase of mature neurons persists as the
G0 phase throughout the remainder of the life of the cell because
further cell division does no occur.
 LIVE - THE ORGANISM
Protoplasm:
 The complex substance (organic and
inorganic) of which all cells are composed.
 Protoplasm exists solely in the form of
organisms and could only be spoken of in
term of living things.
 The essence of the principle of evolution
states that all organisms have arisen from
common ancestry through a gradual process
of change and diversification.
 However, it is generally held that the physical
and chemical composition of the earth’s
surface and atmosphere are no longer
amenable to the de novo creation of life, it
therefore follows that all present day
organism have arisen from preexisting
Individuality:
 Vast differences between individuals in any given
population.
 Differences are due mostly to genetic
constitution:
that is, among the individuals of a given
population, two or more alleles occur at a large
proportion of the gene loci
i.e., although same numbers and kinds of genes
in their chromosomes, the genes may occur in
many alternate forms.
Variations residing in the gene pool of a population
provide the potential for evolutionary change.
The Gene:
 Consists of DNA, an organic molecule composed of two long,
twisted chains of structural units called nucleotide.

 Each nucleotide unit is composed of the sugar deoxyribose,

phosphoric acid, and a nitrogenous base of which there are four
different kinds; viz: adenine, thymine, cytosine, and guanine
(A,T,C,G).

 Therefore there are four different kinds of nucleotide.

 The nitrogenous bases in the nucleotide can fit only in four

combinations: A-T; T-A; G-C; C-G.

 The order of these base pairs varies, and the specificity of any
part of the DNA molecule, depends upon this order.
 Definitions
 Chromatin- replicated DNA and assoc. proteins
 Chromatids- condensed chromatin
 Chromosomes- two sister chromatids attached by
centromere
 Kinetochore- protein band along chromosome
around
centromere
 Mitotic spindles-
 Kinetocore- fibers from centrosome attaching to kinetocore
 Polar- fibers extending centrosome to opposite centrosome
 Astral- fibers radiate outward from centrosome
Mutation:
 Any changes in the code of nucleotide pairs in the DNA
molecule will result in a corresponding change in the
protein it codes and, by extension, in the cells and even the
organisms of which the protein is a part.

 Such alteration of the DNA molecule constitutes mutation.

* The number of alleles at a gene locus is increased only
through mutation.

Mutations are the sole source of new genetic variability and

therefore lie at the root of evolutionary potential. It is
genetic variability within the gene pool that is the raw
material of evolution.

For instance, fertilization, which is the union of reproductive

cells, provides for a new and unique combination of genetic
propensity.
 Primary Terms in Embryogenesis:
 REF: glossary – back of the text book:

 *Embryology – the study of the embryo.

 *Ontogeny – The developmental events or history of an

individual that bring it into being: the totality of developmental
operations.

 *Embryogeny – origin of the embryo: the segment of ontogeny

that precedes hatching or birth.

 *Phylogeny – the succession of forms that culminates in any

given structural entity is the evolutionary history or phylogeny
of that entity. The term applies to the total body form or any
part thereof.
 *Homologous and Analogous – refer to structural
and functional similarities:

 Homologous – equivalent parts in the sense of

having a common ancestry and regardless of
structure and function are said to be homologous, or
are homologues.

 Analogous – parts having similar functions whether

homologous or not, are said to be analogous, or are
analogues.

 Early embryonic stages of vertebrates reveals close

likeness to fully formed fishes, amphibia and
reptiles. However, as development proceeds the
embryos of these different animals become more
and more dissimilar
 The Vertebrate
 Evolution and Development

 Modifications of development  new features specific to

classes/species
 THE VERTEBRATE
THE CHORDATE - features:
 All vertebrates are chordates; but not all chordates are
vertebrates.

 Vertebrates share certain features with some non-

vertebrates - features possessed by all chordates:

n Notochord – lies along the dorsal midline; provides axis

support for the body; present in embryos of all chordates:
persists throughout life in some forms: In most chordates,
notochord is transient & is replaced by the vertebral
column.

n Hollow neural tube – dorsal and above the notochord;

the central nervous system of chordates.
n Pharyngeal clefts (slits) – passageways from the
pharyngeal portion of the digestive tract to the exterior:
- respiratory organs (gills)carries over to adult life in
aquatic
chordates, and serves as
- Only transient existence in non-terrestrial chordates

n Sub-pharyngeal gland – not as salient a feature as (1-

3);
- Located ventrally in the pharynx;
- binds iodine and related substance:
- called endostyle in primitive chordates:
- in vertebrates, its homologue is the thyroid gland.

In addition to these exclusively diagnostic

characteristics, most chordates also exhibit the
following features:

n Tail – rearward extension of the terminal opening of the

digestive
tract.
n Liver – lies ventral to the digestive tract, and kidneys
(distinct tubular unit) rest dorsal to the digestive tract
n Heart – ventral to the digestive tract; pumping blood via
a closed system of vessels.
n Internal skeleton (over and beyond the notochord) –
provides support and protection.

 Other features shared with other

animal phyla and are thus not
exclusively diagnostic:

n Bilaterally symmetrical body – right and left are mirror

images of each other.
n Metamerism or segmentation – serial
repetition of such structures as nerves, blood
vessels, muscles and certain other parts.

n Coelom – all chordates have a true body cavity,

or
coelom.

 Fig 2-2 reveals the diagnostic characteristics of

a true vertebrate (FIG-2).

 * All of the non-vertebrate chordates are

commonly referred to as PROTOCHORDATES.
 CLASSIFICATION OF VERTEBRATES

A. Mammalian
-muscular diaphragm for breathing;
-a four-chambered heart;
-a single aorta;
-two occipital condyles;
-nourish young by lactation
-has functional placenta (except in the egg-laying
mammals) to nourish the fetus and remove waste products from
it.
A. Aves
- 4-chambered heart
- endotherm (warm-blooded)
- attainment of high and constant metabolic rate
- internal thermogenesis  heat is derived from within
the body,
- characterized by feathers and adaptations for flight
- - uses yolk in eggs to nourish the young/embryo
C) Amphibia
- for the most part semi-aquatic;
- are ectothermic.
- mostly egg-laying
- Includes: frogs, toads, salamanders.
B. Pisces
– various classes of fishes
- all built around a common blueprint as a superclass
Pisces.
4 major classes of the Pisces:
1. Agnatha - jaw1ss ; extinct except for the lampreys and
hagfhes
2. Placodermi – armored , jawed fishes;
3. Chondricthyes – modern fishes, the sharks, skates and
rays;
4. Osteichthyes – advanced bony fishes which constitute
half the living species of vertebrates.
 overview

 THE VERTEBRATE:
 Pisces amphibia  reptilian aves 
mammalia
 Are chordates –> identical anatomic features
 Same/common developmental process
 EMBRYOGENY
 Since all animals are related in some degree, it is
possible to sketch an outline of the early stages
of development that applies to all classes of
vertebrates.

The basic events involved in development are:

n Gamete maturation –
gametogenesis
n Fertilization
n Cleavage
n Gastrulation
n Organogenesis  organ system
 overview
n Gamete maturation :
- gametogenesis spermatozoa + oocytes (both haploid)
- a period of chromosomal reduction from diploid to
haploid in both spermatozoa and ova.

n Fertilization
- gametes  zygote (unicellular/diploid)

fertilization initiates two events viz:

– Activation of the ovum to start dividing i.e. initiates
development, and
– Establishment of diploidy - union of the two haploid
gametes.
 overview

c) Cleavage
- a period of segmentation of the zygote
- series of mitotic divisions blastula (multi-cellular)

d) Gastrulation
- further mitotic divisions of blastula  gastrula
establish primary germs layers:
ectoderm (outer);
endoderm (inner);
and mesoderm (middle).

e) Organogenesis  organ system formation

EMBRYONIC ORGANIZATION:
 The Ectoderm - the outer strata of the embryo
represents the embryonic skin.
 The Endoderm - inner stratum is the future lining of
the
GIT;
the space it encompasses is the cavity of the GIT, the
archenteron.
 TheMesoderm - between skin and embryonic gut;
 Neural Tube -ectodermal in origin; along dorsal axis of the
embryo;
formed by invagination of the neural plate and joining of
the neural folds in the midline;
; anterior portion will form the brain; the remainder will
form
the spinal cord of the adult.
 overview
 The neural crest
- arise from cells within the neural folds;
- and occupies either side of the neural tube;
- derivatives include - ganglia of the cranial and spinal nerves:
- some of the cells migrate to parts of the body to form
different tissues
including the sympathetic nervous system (SNS), sensory
ganglia of
cranial nerves V, VII, IX and X; adrenal medulla, pigment cells;
teeth
parts of the skull.
 The neural crest
 Is Ectodermal in origin
 Adult derivatives:
- ganglia of the spinal nerves
- sympathetic nervous system (SNS),
- sensory ganglia of cranial nerves V, VII, IX and X
- adrenal medulla
- teeth
- parts of the skull.
- pigment cells
 The Notochord and Mesoderm
- notochord is derived from mesodermal cells that
segregate from
median dorsal mesoderm; lies between the gut floor and
the
developing neural tube above.

The lateral mesoderm spreads ventrolaterally and the spreading

sheets on either side meet in the ventral midline.

At same time these mesodermal sheets split into two layers:

– splanchnic mesoderm and
– somatic mesoderm
 overview

 The cavity between them is the coelom - a true cavity

bounded by an epithelium of mesodermal origin.

 The splanchnic mesoderm unites with adjacent endoderm

to form the splanchnopleure.

 The somatic mesoderm unites with ectoderm to form the

somatopleure .
 Dorsoventral division of the
mesoderm into:
n Epimere - most dorsal and flanks the neural
tube and notochord; gives the following
derivatives – sclerotome, dermatome and
myotome;

n Mesomere - narrow; between the epimere and

hypomere;
n Hypomere - most ventral and flanks the gut.
Note:
 When we speak of an organ or part of it being derived from a
given germ laver, we have in mind only the intrinsic
functional portion of that organ; for there is no structure in
the animal body that is the product of a single germ layer
exclusively.

For illustration:
e) The intestine is considered to be endodermal in origin, yet
only its secreting and absorbing interior lining is so derived.

b) The muscles, peritoneum, connective tissues, blood vessels,

and nerves, which constitute the bulk of the GIT, are derived
from mesoderm and ectoderm .
 overview

a) In normal development there are instances in which the

cells of one germ layer give rise to structures customarily
farmed by those of other germ layers.

c) The developing tail of vertebrates is illustrative.

e) The muscles of the tail are a product of ectoderm,

whereas muscles in general are mesodermal in character.

g) Further, the skeleton is ordinarily derived from

mesodermal connective tissue,

i) yet the gill skeleton is produced by neural crest cells

originally associated with the nervous system.
 * Yet the organ-germ layer concept is still useful as a
descriptive scheme of classification and identification.
Furthermore, there is a high degree of homology between
these layers and the products provided by them.
What are the Components of Development?

 Development
- a series of changes culminating in the formation of a
complex
organism from relatively small and simple, one cell
organism. Includes:
i) Growth; ii) morphogenesis; iii) differentiation
 overview

n Growth - developmental increase in mass (synthesis

of new protoplasm - cytoplasm, nucleic and cytoplasmic
products) accompanied by cell division which is
characteristic of growth.
n Morphogenesis - generation of new form; the
organized movements of multiplying cells and the
attendant physical reshaping of areas. A sheet of cells may
undergo delamination (splitting) into two or more layers; a
cord may hollow out (undergo cavitation); there may be
folding, invagination (inpocketing), evagination
(outpocketing).
n Differentiation - encompasses a host of
operations culminating in increasing diversification of form
and function. Events by which cells and other parts become
different from one another and also different from what they
were originally.
 * Morphological differentiation - as
they multiply, cells become structurally different from other
cells. E.g. from a general ectoderm --> nerve cells and
epidermal cells acquire distinguishing features of size,
shape, and internal architecture.
* Behavioral differentiation -
acquisition of special functional capabilities: e.g. nerve cells
come to transport electrical disturbances; muscles to
contract; gland cells to secrete special products. etc.
* Chemical differentiation - as an
illustration, consider the egg. An egg has complex chemical
composition. As it cleaves and progressively become a
blastula, gastrula and embryo, individual cells and areas of
cells become biochemically different from one another. The
process by which this fixity of developments prospect is
acquired is called DETERMINATION.
 overview

 Differentiation - could therefore be redefined

as the series of events by which materials become
chemically determined and then assume distinctive form
and function.
THE GAMETES:
 In all vertebrates reproduction is effected by
sexual means with the use of specialized
germ cells called gametes - spermatozoa in the in
.
male and ova in the female

 The Primordial Germ cells: There are different concepts

about the origin of the germ cells found in the adult gonads.
n One doctrine states that the PGCs are merely special
endodermal wandering cells, having nothing to do with
reproduction;
and that gamete formations is a matter of
differentiation of somatic cells within the gonads.
i. Another doctrine:
- PGCs are extragonadal cells gonads gametes.

 In vertebrates, identification of the PGCs comes later in

development ;  are external to prospective gonads
and external to embryo when first recognized

 Most recent notion:

 - PGCs are the sole progenitors of the gametes.
 How do the PGCs reach the presumptive gonads?
iv. Extra-vascular/mesenteric route:
The PGCs may migrate, following cell surface signals
such as
fibronectin in the dorsal mesentery and peritoneum
into the
region of the developing gonads (genital ridge).
ii. The vascular route:
PGCs may go through the vascular system to the
genital ridge.
- birds and reptiles
 Developmental Period Stages
A.Embryogenesis
 Stages of embryogenesis:

fertilization  cleavage 
gastrulation  organogenesis
fertilization  histogenesis
(differentiated tissues)
 overview
n Fertilization - union of egg and sperm.
Classification of egg: micro-, meso-, and macro-
lecithal.
One polarity axis - animal-vegetal axis.

 Fertilization establishes:
Stimulation of egg
Diploidy (2n) Fig 1.6

9. Cleavage
Definition
Classification
Blastomeres
End result of cleavage – Fig. 1.7
n Gastrulation
- involves movement of cells.
- Morphogenetic movements - cells rearrange; migrate;
spread;
bend and fold forming a
gastrula.
- End-product of gastruIation  3 germ layers.
ectoderm
mesoderm
endoderm
 READING – text book:
Chap 12
- migration of the germ cells
- fertilization
- gametogenesis spermatogenesis +
oogenesis
 Gametogenesis
 The totality of preparatory events
leading to the formation of the haploid
gametes from diploid gonials through
the process of mitotic and meiotic
(reduction) divisions.
 Spermatogenesis
 Occurs in the
seminiferous
tubules in the testis
A. Spermatogenesis:
i Spermatocytogenesis - involving mitotic and
meiotic divisions of the spermatogonia through
formation of spermatocytes and spermatids.

ii Spermiogenesis (spermateliosis) -
morphogenesis of the spermatid to yield the
characteristic spermatozoa ; no cell division
involved.

iii Spermiation - release of spermatozoa into the

lumen of the seminiferous tubules

iv Emission

 spermatogenesis
Spermatogonia

Spermatocytes

–primary

- secondary

Spermatids

- round

-elongated

Spermatozoa
 The Spermatozoa
 Sperm - great morphological diversity between kinds of
vertebrates.

 Head and tail (neck-midpiece-principal piece/tail-endpiece).

 Head - spheroidal in teleosts; rod or lance-shaped in

amphibia; spirally twisted in passerine birds; spoon-shaped
in humans and many other mammals; hooked in mouse and
rats.
 Serves two functions: genetic (in the DNA), and activating.
Mitosis and Meiosis
 Spermatogenesis
 Spermatocytogene
sis
 Spermiogenesis
 Spermiation
 Emission
 Ejaculation
Spermatogenesis
 Spermatogenesis
starts in testes in
seminiferous
tubules
 Spermatogenesis
starts in outermost
layer of tube and
daughter cells
move towards
lumen.
Male Reproductive Tract

Sperm pass
through

 Seminiferous
tubules
 Epididymis
 Vas Deferens
 Ejaculatory
Duct
 Urethra
 Penis (within)
 Seminiferous Tubules
 In testes
 Very convoluted: if
stretched, 1.5 miles!
(in one testicle only)
 Sertoli cells=
Sustentacular cells:
sustain sperm
 Leydig cells=
Interstitial cells: make
androgens
Functions of Sustentacular
Cells

 Secretion of Müllerian-inhibiting factor

 Support of mitosis and meiosis
 Support of spermiogenesis
 Secretion of androgen-binding protein
 Secretion of inhibin
 Maintenance of blood-testis barrier
Seminiferous Tubules
Spermatozoon
 Head
 Neck
 Tail
 Lacking many
intracellular
structures
 At this step, can’t
fertilize or move
coordinately
A. Oogenesis:
 The totality of the processes involving
mitotic and meiotic divisions of the diploid
oogonia, culminating in the formation
haploid ovum, and nonfunctional polar
bodies.

The Vertebrate Egg:

 The nutritive substance in all eggs is the
yolk. Yolk is primarily produced in the liver,
and is transported in a soluble form via the
bloodstream to the ovary.
 In the ovary, the yolk is transferred by
ovarian cells that surround the oocyte and
is deposited as platelets or granules. The
vertebrate is classified according to
amount of yolk in the cytoplasm.
Oogenesis
Step 1

Oogonia are small cells

that increase in number
by mitosis
When they enlarge
slightly, they are
considered primary
oocytes
 Primary Follicle
Oocyte enclosed by Follicle
cells
Primary oocytes become
enclosed by a single layer of
follicle cells
Follicle cells from the
surface layer of the
ovary
Primary follicle is the
structure including a primary
oocyte enclosed in follicle
cells
 Oogenesis
At puberty, the primary follicles enlarge
rapidly and accumulate yolk caused by
the
follicle
stimulating hormone (FSH) from the
pituitary gland

As the yolk increases, the oocyte is

pushed to one side as a small disc
The disc is called the blastodisc

The blastodisc contains the enlarged

nucleus of the primary oocyte
which is called the
germinal vesicle

YOLK

BLASTODISC
 Oogenesis
 First meiotic division
of the primary oocyte
occurs just before
ovulation
 The result is the first
polar body pinching
off and the secondary
oocyte
 The secondary oocyte
contains most of the
yolk and the
blastodisc
Fertilization
The second meiotic
division begins but stops
at metaphase
The secondary oocyte
(sometimes called the
ovum at this stage) is
mature enough to be
fertilized and
Ovulation occurs
 Oogenesis/Fertilization
 The secondary oocyte
(ovum) is released
from the ovary as the
follicle ruptures
 It enters the
infundibulum of the
oviduct where it will
encounter sperm cells
 As sperm penetrates
the blastodisc, meiosis
begins again and the
result is the second
polar body and the
mature ovum
 READING– text book
Chapter 8
- fertilization
- cleavage  amphioxus
xenopus
chick
sea urchin
- gastrulation
 Cleavage and Blastulation
 Cleavage- Occurs in eggs activated
by fertilization. Cleavage is the
subdivision of eggs into cells called
blastomeres. The blastomeres divide
quantitatively into smaller units until
they are of such a size that they can
readily undergo the subsequent
events of blastulation, gastrulation,
and interaction that are involved in
formation of tissues and organs
 Cleavage & Blastulation
 The blastodisc initiates
cleavage
 First cleavage occurs
in the zygote by way
of mitosis and the
result is two cells
called blastomeres
 Cleavage occurs in the
cytoplasm, not the
yolk
 Second cleavage = 4
blastomeres
 Cleavage (mitosis)
continues until the
blastula forms
Blastula
Blastulation
•The formation of a segmentation cavity or
blastocoele within a mass of cleaving
blastomeres. Rearrangement of blastomeres
around this cavity forms the type of definitive
blastula, characteristic of each species
•The definitive blastula is thought to terminate
cleavage stages
•The wall of the blastula is a mosaic of cellular
areas, each of which will normally produce a
certain structure during subsequent
development. In other words, each area of cells in
the wall of the blastula has a certain prospective
fate which will be realized in normal development.
 The Vertebrate
Egg:
Nutritive substance in all eggs is the yolk.
Yolk is produced in the liver  transported in a soluble form via
the bloodstream to the ovary.

 In the ovary, the yolk is transferred by ovarian cells that

surround the oocyte  is deposited as platelets or granules.

 Classification of egg:
The vertebrate is classified according to amount of yolk in the
cytoplasm:
microlecithal: mesolecithal: macrolecithal
 Eggs:
n Microlecithal - small amount of yolk, scattered
evenly in cytoplasm (Amphioxus; tunicates;
eutherian mammals)
n Mesolecithal - relatively larger amount of yolk
in cytoplasm; not quite evenly scattered
(Amphihia; dipnoi; Petromyzontia).
n Macrolecithal - enormous amount of yolk as
food reserve (Myxinoidea; Chondrichtyes;
Osteichthyes;reptiles; and birds).
* Telolecithal eggs (mesolecithal and
macrolecithal) - where yolk is concentrated in one
hemisphere the vegetal pole of the egg
(greatest concentration of yolk),
than the animal pole (smallest amount of yolk).

 * The amount and distribution of yolk influences

the pattern of cleavage and movement of cells
and tissues during gastrulation.
 Fertilization
 Union of the ovum and spermatozoon to form the
zygote.
 Occurs in fluid medium - All the events involved in
fertilization (internal
and external) take place in a fluid medium.

 Barriers encountered by the spermatozoa include:

cumulus oophorus; corona radiata (encasing the ova); zona
pellucida;
the vitelline membrane; mucus in the genitalia (internal
fertilization).
 In some vertebrates (mouse) the zona pellucida contains
glycoprotein (zona glycoprotein) which acts as receptor for
the binding protein of the sperm membrane.

 The acrosome produces hyaluronidase (lytic enzyme) which

dissolves the egg barriers.

 The acrosome of many different kinds of sperm have been

shown to produce a specific protein called bindin, which
reacts with bindin receptors (glycoprotein) on the ovum’s
vitelline membrane on contact, thus attaching the tip of the
acrosome to the membrane.
Acrosomal Reaction:
 Morphological changes and biochemical events undertaken by
the spermatozoa in order to acquire fertilizing capability.

 Identified in several vertebrates and invertebrates.

 Capacitation: Sperm of several species require a period of

exposure to compounds - glycosaminoglycans (GAGs) in the
female genitalia before the bindin reaction can occur.
This preparation is part of what is known as sperm
capacitation .
 Steps in AcR
n Disappearance of the Acrosomal membrane at the
apical region.
n The Acrosomal and sperm membranes become
continuous (Fig 5-2A).
n At the same time, disintegration of Acrosomal
granule, as does the portion of the egg membrane
exposed to the granule (signifying release of lytic
enzymes).
d) Indentation of that region of the acrosomal membrane
abutting
the nucleus (arrow in 5-2A).

e) Elongation and deepening of the indentation, thus

creating a
steady elongating Acrosomal filament that traverses the
egg envelope and finally approaches the plasma
membrane of the ovum (5-3C).
f) Zygote Formation:
Establishes definitive contact between the two gametes
(with the meeting of the sperm and egg plasma
membranes - one continuous membrane. The contents
are now essentially components of one cell - the zygote.
b) Activation of :
Contact of the acrosome with the plasma membrane of the
egg
activates the otherwise quiescent egg into action:

n Polyspermy: Definition of: 

–
Significance of :
Blockage of polyspermy:
Block polyspermy:

a) Fast block to polyspermy:

- contact between acrosome and ovum membrane results in
electrical charges  change in membrane potential across the
surface of the egg, immediately preventing further sperm
penetration of the egg.

b) Slow block to polyspermy:

- cortical reaction; I
cortical vesicles  discharge cortical granules to fuse with
egg plasma membrane chemical alteration in the membrane I 
inactivating its bindin receptors releasing any other sperm that
had attached to it thus preventing further fertilization.
This constitutes the slow block to polyspermy.
i. Consequences of sperm penetration:

a) Establishment of the plane of bilateral symmetry

sperm penetration of ovum  rearrangement of its
internal
constituents  leading to the establishment of the
plane of
bilateral symmetry of the embryo to come.

b) Alterations in the physiological properties of the egg;

changes in permeability ;
and alterations in the rate of metabolism;
ovum becomes a truly dynamic entity and embarks on

physicochemical transformations that lead into the

period of
cleavage.
Establishment of Diploidy - union of the haploid nuclei:
 Fertilization establishes/restores the diploid number of chromosomes for
the new organism.
 Fertilization restores diploidy which represents a new combination of
hereditary prospects.
 In mammals and most other vertebrates, each pronucleus (male and
female) loses its membrane, and concomitantly its chromatin resolves
into a haploid set of chromosomes.
 The two set of chromosomes arrange themselves across the spindle.
 This arrangement, marking the readiness for the first cleavage division,
represents completion of the process of fertilization.

 * Fertilization is completed with formation of the zygote and

initiation of cleavage.
Stages in the Events of
Fertilization
 Fertilization-sequelae - Summary
 1. Prevention of polyspermy via:
- fast: changes in electrical membrane potential
- slow: cortical reaction (granules fusing with egg
membrane

2. Establishment of diploidy

3. Establishment of plane of bilateral symmetry in

presumptive
embryo

4. Establishment of dynamic entity of the ovum 

physiological
transformation initiation of cleavage
 PARTHENOGENESIS
 Fertilization of the ovum to the zygote without spermatozoa.

 This indicates that activation and nuclear phases of fertilization are

essentially distinct; that development does not absolutely require a
union of egg and sperm.

 The eggs of every animal group, including mammals, can be started

on a course of development, often to the completion of a new
individual, by a host of chemical and physical agents.

 This strongly suggests that the ovum possesses in itself all the
capacities to form an embryo, needing only some agents to trigger
the action.
 In normal development, the spindle for the first cleavage division
is assembled around the centriole brought by the sperm.

 What is the source of the centriole in parthenogenesis? This may

reside in the egg. In some cases, spindle appear to arise
spontaneously.

 The easiest way to induce parthenogenicity in the frog is to stab

the egg with a very fine needle. A method that is effective only if
the needle carries

 some foreign protein into the egg. The protein provides a center
around which the cleavage spindle arises
 Although parthenogenesis occurs naturally in some vertebrates (some
birds and amphibians), it rarely occurs in others such as the whip-tailed
lizards (Cnemidophorus uniparens), all the adults of which are females.

 Recent experiments have shown that elements of both male and

female genomes are necessary for full development in mammals.

 Expectedly, the chromosome numbers in parthenogenetic individuals

are haploid.

In some cases, there is tendency for the restoration of diploidy via:

fusion of haploid cleavage nuclei prior to cell division; fusion of egg
nucleus with a polar body nucleus before cleavage begins.

 Advanced or complete development occurs only if regulation to

diploidy is accomplished: haploid individuals generally succumb in
early embyonic stages.
 Cleavage
Eggs – Types:
 Isolecithal - small amount of yolk evenly
distributed; mammals.
 Mesolecithal - moderate yolk in vegetal pole;
amphibians.
 Macrolecithal – large amount of yolk in the
egg
or
 Telolecithal - large yolk filling almost entire
egg; fish, reptiles, birds.
 Centrolecithal - yolk concentrated in the
central cytoplasm; insects, arthropods.
 Cleavage Pattern
 Holoblastic - entire egg cleaves during
cytokinesis.
– Complete Holoblastic  blastomere.
– Partial Holoblastic  micromeres +
macromeres
 Meroblastic - characterized by partial
cytokinesis;
two types:
 Discoidal cleavage - restricted to animal
pole,
yolk-free area.
*Vegetal pole remains uncleaved in telolecithal
eggs.
 Amphibian Cleavage
 From 128 cell stage = blastula
 Several layers thick;
 Larger blastomeres in vegetal pole – macromeres.
 Smaller blastomeres at animal pole – micromeres.
Mammalian Cleavage
Cleavage Cellular Mechanism
 Cleavage Cellular Mechanism

Experimental evidence:
 In isolecithal eggs - mitotic spindle is centered -
cleavage furrows form all around the
circumference.

 In mesolecithal eggs - mitotic spindle is displaced

to the animal pole - the furrow first appears in
animal pole then cuts towards the vegetal pole.

 In telolecithal eggs - mitotic spindle is eccentric -

the cleavage furrow forms only near the mitotic
apparatus.
 Cleavage & Gastrulation
Cleavage - general consideration:
 Series of a regular rhythm of mitotic divisions of the egg following
activation either by the spermatozoon or by parthenogenetic agent,
and ending with formation of a blastula.

 The individual cells during the process of segmentation are called

blastomeres.

 During the period of segmentation there is no increase in cytoplasmic

mass (embryo does not grow); segmentation leads to increase in the
number of blastomeres but not the size;
; there is no growth during these mitotic divisions as the G1 and G2
phases of the cell cycle immediately before and after DNA synthesis
are eliminated.
 With each cell division there is additional DNA synthesis
and protein, to provide for the increased nuclei,
at the expense of cytoplasmic reserve  nuclear gain is
balanced by cytoplasmic loss:
 leading to numerous small blastomeres each with its
nucleus and cytoplasm.

 Since the area of constriction that divides the cytoplasm

(cytokinesis) is organized by the centriole of the cell, the
plane of cell division is always at right angle to the long axis
of the spindle
Variations in Cleavage Pattern:
b. Amount of yolk in egg cytoplasm
 Influences the rate and pattern of cleavage
division; the greater the amount of yolk and the
more unevenly distributed the yolk, the more the
mitotic spindle in the egg and later in the
blastomeres are displaced from the center of the
cell toward the less yolky ends; and upon division,
blastomeres of unequal size result. In an egg with
most heavily laden yolk (birds), segmentation is
confined to a small region (blastodisc) at the
animal pole.
a. Cytoplasmic organization
 Exemplified by the spiral cleavage in annelids
and mollusks - where the rotational movement
of blastomeres resulting from alternately tilted
obliquely to the right and left, so that
successive generations of blastomeres are
oriented in a twisted fashion.
Summary – cleavage pattern
n Holoblastic (total) Cleavage:
 The whole egg divides as do the blastomeres:
– Equal holobalstic: In microlecithal eggs (mammals)
- blastomeres of equal sizes
– Unequal holoblastic: In mesolecithal eggs
(amphibians) – blastomeres of unequal sizes
n Partial Cleavage: (Teleost fish) part of egg
remains undivided; in moderately macrolecithal
eggs

n Meroblastic Cleavage: (reptiles; bird &

primitive mammals) - extremely macrolecithal
eggs –segmentation is confined to small area at
the animal pole.
GASTRULATION – commences at the conclusion of
segmentation & formation of the blastula
Gastrulation -
 GASTRULATION
 is by a process of rapid cell multiplication by which the blastula
is molded into a stratified structure - gastrula

 involves flattening and invagination (inward folding) of the

 prospective endodermal plate into the blastocoel

 the new cavity is called gastrocoel (archenteron) which opens

to the exterior by the blastopore
 further migration of cells and elongation of the
gastrula establishes two layers: a) an outer
ectoderm, neural and epidermal, and b) inner cell
mass encompassing prospective notochord,
mesoderm and endoderm

 the organs of the body are all derivatives of these

three primary germ layers.
 Note:
 In amphoxius, the mesodermal pouches are hollow from the beginning
which is to say that they contain a coelum.

 Thus the Amphioxus features an enterocoel, (the coelomic spaces

trace their origin to the cavity of the primitive gut).

 The vertebrates feature a schizophoel - a coelom formed by the

splitting of an originally solid layer of mesoderm.

 In each case the end result is the same: somatic mesoderm associated
with epidermal ectoderm to form somatopleure; and splanchnic
mesoderm associated with endoderm (endodermal gut) to form
splanchnopleure.
 AMPHIBIAN - FROG
 eggs are mesolecithal - moderate amount of yolk present in
form of oval granules scattered throughout the cytoplasm,
but show gradual concentration to one pole;
 telolecithal - greater concentration of yolk located at the
vegetal pole.
 Cleavage: follows the pattern seen in the amphioxus with
minor differences:
  Gastrulation (cont’d)
 involves invagination, involution and stretching and spreading
of cells of the blastula to form a spherical gastrula surrounded
externally by ectoderm and containing endodermal and
mesodermal components internally, comparable to the situation
in amphioxus.

 throughout gastrulation the embryo retains its spherical shape

and a uniform size.
 endodermal ridges meet in the dorsal midline and unite to
create a tubular endodermal gut.

 mesoderm in the dorsal midline is set off as a definitive

notochord, separate from the presumptive epimere,
mesomere and hypomere the forerunners of other organs.

 the overlying ectoderm would give rise to neural tube.

 Avian
 first cleavage division is an irregular shallow furrow in the
central portion of the blastodisc.

 this furrow does not cut completely through the depth of the
disc; and this results in two incompletely separated
blastomeres.

 the second division occurs at right angle to the first but does
not cut entirely through the depth of the disc.
 further incomplete divisions result in irregular pattern of
cleavage, creating two groups of blastomeres: a)
completely bounded central blastomeres and b)
incompletely bounded marginal blastomeres.

 further divisions of the blastomeres create a stratification of

cells. This results in a compact plate several cells thick;
which is called the blastoderm.

 the bulk of the yolk is external to the blastoderm (which still

contains some yolk cells).
 The bilaminar blastoderm represents the blastula.; the epiblast
represents the animal half; and hypoblast (endoderm)
represents the vegetal half; and the intervening space,
represent the blastocoel.

 Gastrulation
 the hypoblast (endoderm) is already present in part at
beginning of gastrulation.

 Major differences in the fate of embryonic cells of avian and

amphibian?
 i) formation of extraembryonic organs from the blastoderm
to support embryo prior to hatching

 The epiblast of the entire area opaca is destined to provide

extraembryonic ectoderm.

 The epiblast (upper layer) is the presumptive ectoderm,

mesoderm, and intraembryonic endoderm.

 The anterior two-thirds of the area pellucida is prospective

ectoderm.

 The greater part of the ectodermal area is prospective

dermis and the bulk of this is extraembryonic.
 ii) Invagination in the frog involves folding and
retraction of a sheet of cells into the
blastocoel, formation of archenteron and a
blastopore: in the avian, the separation of
these layers from the ectoderm involves the
process of convergence and ingression.
 the process of convergence and ingression initiate the
arrangement of mesodermal and endodermal tissues
between the epiblast and hypoblast

 the final stages of gastrulation in the avian embryo involve

a continuation of convergence and ingression of cells
culminating in the establishment of the germ layers and
other presumptive organs- somite and epimere, notochord
etc.
 Placenta formation in amniotes
 *The four extraembryonic membranes are:
 i) amnion - protective
 ii) chorion - respiratory and nutritive
 Iii) allantois - respiratory and nutritive
 iv) yolk sac - nutrition (source of albumin)

 Salient points:
 a) Amniota: - amnion-possessing vertebrate such as:
mammals: birds and some reptiles .
 b) ANAMNIOTA: - cyclostome, fishes, and amphibian only
EEM is the yolk sac (lacking other membranes including the
amnion.

 c) OVIPAROUS:- external deposition of eggs and young

developing independent of parents.
 all birds; most fishes, amphibians and reptiles

 d) VIVIPAROUS. - retention of egg by a parent (in female

reprod. tract); or specially designed pouch in male or
female,) ; and young are “born alive”.
 THE REPRODUCTIVE CYCLE
The ovarian cycles in mammals
shedding of blood and cellular debris from the uterus at montly
intervals. The menstrual cycle represents the integration of three
very different activities:
(1) the ovarian cycle, the function of which is to mature and
release an
oocyce,
(2) the uterine cycle, the function of which is to provide the
appropriate
environment to the developing blastocyst to implant
(3) the cervical cycle, the function of which is to allow sperm to
enter the
female reproductive tract only at the appropriate time.
These three functions are integrated through the hormones of the pituitary,
hypothalamus, and ovary.
 The majority of the oocytes within the adult human ovary are
mainly in the prolonged diplotene stage of the first meiotic
prophase (often
referred to as the dictyate state. Each oocyte is envelopped by a
primordial follicle consisting of a single layer of epithelial
granulosa cells and a less-organized layer of mesenchymal thecal
cells

Periodically, a group of primordial follicles enters a stage

of follicular growth. During this time, the oocyte undergoes a 500-
fold increase in volume (corresponding to an increase in oocyte
diameter from 10 micro-meter in a primordial follicle to 80 micro-
meter to a fully developed oocyte.
 the secondary stage of follicle development is marked by the
appearance of the zona between the granulosa cells and the ovum.

 antrum: a follicular cavity is formed by creation of a cleft within the

granulosa cells; this is called antrum

 tertiary follicle; a follicle with an antrum is called tertiary follicle

 further increase in size -, antrum enlarges, oocyte still attaches to
granulosa on one size.
 cumulus - the granulosa investing the ovum is now called cumulus.
 theca cells - as the follicle grow, a framework of connective
tissue is laid around the ovum, outside the granulosa cells:
this is the theca layer outside the parietal granulosa.

 oocyte, still encased in the zona pellucida and remnants of

the cumulus, stretches and floats in the antral fluid; the
release of the cumulus from the theca-granulosa causes
them to extend out into the follicular fluid in a mass of
projections called corona radiata.
 Ovulation:
 the mature follicle, called the Graafian follicle, finally bursts open to
release the oocyte. This process is called ovulation.

 in most mammals ovulation is followed by completion of the first

meiotic division and extrusion of the first polar body.

 in uniparous mammals, such as the human female, only one follicle

develops into maturity and only one follicle ovulate, generally.
 in litter-bearing mammals (pluriparous) several follicles
attain maturity, and several ova are eleased at ovulation.

 * Considerable variability in timing of ovulation in mammals

may occur hours before or after heat: in human female
ovulation may occasionally occur almost a any time in the estrus
cycle.

 Ovulation is not spontaneous in all mammals. In rabbit, cat, mink

ovulation takes place only after nervous stimulation derived from
the act of copulation.
- these are called induced-ovulators
 The Estrus Cycle:
 the breeding pattern and behavior among the vertebrate are
remarkably variable.
this is dependent on whether or not the breeding cycle is seasonal or
intra-seasonal.
Some mammals mate periodically throughout the year;
others mate only during a restricted breeding season.
except the higher primates, all female mammals permit mating only
at definite times called “heat” or estrus.

 Estrus = a period in the estrus cycle during which the female mammal
is in a physiological and psychological readiness for mating, and hence
sexually accepts the male.
 Repetition of the estrus period at a set interval constitutes the
estrus cycle.

Estrus cycle length; - varies remarkably in different vertebrate

groups:

 Rat and Mouse - 4 to 5 day interval

 Dogs - every 6 months (twice yearly)

 Domestic cow and pig - 3 weeks (18 - 25 days)

 Deer family and wild sheep - mating only once a year.

 Phases of the Estrus’s cycle

 a) Estrus = a period of sexual desire and acceptance of mating
 b) Metestrus = a period of preparation for pregnancy - increasing
P4 from
developing CL anestrus  proestrus estrus etc