BIO LEC EXAM 1 Specialized Fields of Embryology

Embryology and Genetics

Descriptive Embryology
Embryology and Genetics  Describes the processes and sequential events of development
 Merging of two sciences that are intricately intertwined to study the which transforms a single-cell zygote into a multicellular organism
production of living organisms  Gives a step-by-step account of the processes of development
 Unique among all other biological sciences, as it seeks to answer  Answers such questions as “What are the processes?” and “How are
the central paradox of life they formed?”
Central Paradox of Life Comparative Embryology
 How can a single fertilized egg give rise to a multicellular organism  Examines similarities and differences in the development of different
like me? vertebrate groups
 1+1=1  Study of how anatomy changes during the development of different
 How can that one cell develop into all the different cell types of the organisms
body?  “Ontogeny recapitulates phylogeny”  Recapitulation
 ~ 250 cell types, all derived from a single diploid cell (2N) Theory/Biogenetic Law by Haeckel
o Ontogeny is the history of the individual
Embryology and Its Scope o Phylogeny is the history of its race
 Von Baer’s Law – more general features appear earlier in
development, than those that are characteristic of a particular group
Embyrology
 Study of the origin and development of an organism from a fertilized  Phylotypic stage – Stage during embryological development wherein
egg to the period resembling an adult form different species seem almost identical
Importance of Embryology
1. One of the very important major courses in the entirety of biology
2. Foundation of modern sciences like anatomy, pathology, genetics,
evolution, cellular biology, and ecology
3. Enables one to understand much better other fields of biology, and
to appreciate higher biology
4. Explain the mechanisms of the development of organisms, which is
mainly responsible for the diversity of animals
5. Answers questions on what makes animals different
 What makes vertebrates different from invertebrates?
o Neural crest cells: highly migratory, undifferentiated cells  Note: Craniates = dorsal nerve cord, pharyngeal pouches, post-anal
perched between the epidermis and the neural tube tail, notochord
o Develop into facial features (cranium, facial muscles) Experimental Embryology
Developmental Biology  Test hypotheses and manipulate by experimentation
 Broader study that deals with organogenesis and postnatal  Pioneered by Wilhelm Roux
development  Involves tissue and organ transplants – excising a portion and
 Development continues on even at adulthood graphing it to another
 Examples of adult development o Homotransplantation involves same species
a. Neoplastic growth – abnormal proliferation of cells o Xenotransplantation involves different species from same
(tumors) due to malfunctioning of control points order
b. Regeneration – regrowth of body parts from pieces of an Reproductive Biology
organism, more prominent in lower forms of organisms  Investigating conception and contraception
such as starfishes, reptiles  Emphasis on the following
c. Metamorphosis – process in which an animal physically o Normal gametogenesis (prior to meeting)
develops after birth, transition from larvae into adult o Transport of gametes and fertilization
d. Tissue repair – repair tissues at levels of complexity o Endocrinology of reproduction – hormones
ranging from the molecular to the organismic level (e.g. o Early embryonic development
blood clotting); scar – tissues that have replaced o Implantation of mammalian embryo
destroyed ones Chemical Embryology
Developmental Genetics  Study of the chemical and physical events in development
 Studies the genetic mechanisms involved in the development of an  Interaction of chemical and physical factors greatly affect
organism development
 Deals with the manifestation of traits coded in genes, and how they Teratology
can be altered to produce amazing results  Study of birth defects
o 6-eyed cat  Deals with abnormalities and malformations in development due to
o Zebroid foal – result of gene-spliced zebra +donkey, 2x genetic events and exogenous factors
bigger than ordinary baby zebra
 How genotype is translated into phenotype
Genetic Exogenous
 Human Genome Project
Mutations Drugs
o International project aimed to determine the totality of the
Aneuploidy – change in chromosome Radiation
human genome
Translocation – change in postion of
o 25,000-30,000 genes comprising the human genome Alcohol
base pairs
o Wellcome Trust Sanger Institute
Bacteria
Viruses

 Affects the main stages in histological development

 o Proliferation
o Migration
o Differentiation
 Phocomelia
o Condition wherein long bones of limbs are either absent
or severly deficient
o Etymology from phoco (“seal”) and melia (“limb”)
o Caused by genetic inheritance and/or mutations due to
radiation and by oral intake of the drug thalidomide
prescribed as mild sedative for pregnant women
o Women should be careful especially in the 1st trimester
Embryology in Contemporary Science
 Test Tube Baby/ In Vitro fertilization – egg and sperm are allowed to
meet in a controlled environment (test tube)
 Embryo transfer – embryos transformed through in vitro fertilization
are transferred to surrogate mothers
 Animal cloning – process of producing genetically identical living
organisms
 Cloned animal is produced from a preexisting animal
 Dolly – first ever cloned animal
 Produced by Ian Wilmut and Keith Campbell through their studies at
the Roslin institute
 Produced from a single mammary gland cell
o Proves that even highly differentiated adult somatic cells
can revert back to its embryonic origin
 Biological clock of cells were not reset causing faster aging
History of Embryology
Aristotle
 “Curiosity is the mother of invention”
 Posed the question “how are the different parts of a developing
embryo formed?”
 Preformation
o Miniature adult-like embryo already existed and only got
bigger
o Spermists
 Sperm cell contains a new individual and is
merely nourished by a ovum
 Encasement Concept: a homunculus, a tiny
human in fetal position in head of a sperm
o Ovists
 Argues that ovum contained a miniature body
which is stimulated to grow by the seminal
gluid
 Charles Bonnet strengthened the ovist thought
by discovering that some insect eggs
developed in female species by
parthenogenesis
 Epigenesis
o Gradual formation of new structures and organs, like
knitting a net
o From a single cell arises a multicellular organism
Galen
 Studied the structure of relatively advanced fetuses
 Development was not studied well since the early stages are the
more critical points of development
 Study restricted by the miniature dimensions of early embryos
(prevented serious analysis)
 Resolved by the development of the microscope in the 17th century
Anton van Leeuwenhoek & Louis Hamm
 First to see human sperms with crude microscope
 From a drop of water they saw bacteria, protozoa, and sperm cells
Regnier de Graaf
 Founder of modern reproductive biology
 Described ovarian follicles and Fallopian tubes
o Mature ovarian follicles are called Graafian follicles
o Studied ectopic pregnancy, wherein the fetus does not
develop within the uterus
Lazzaro Spallanzani
 Italian priest, physiologist, and natural scientist
 Stated that in normal circumstances, male and female sex products
are necessary to initiate development
Marcello Malphigi
 Italian embryologist
 Published the first microscopic account of chick development in
1672
 Unconvinced of epigenesis because the unincubated egg had many
structures
Karl Ernst von Baer
 Russian zoologist
 Developed the science of comparative embryology
 Von Baer’s law: more general basic features of any animal group
appear earlier in development than special features peculiar to
different members in the group
 Germ Layer theory: existence of germ layers in embryos from where
all organs are derived
o Endoderm – gut, liver, lungs
o Mesoderm – skeleton, muscle, kidney, heart
o Ectoderm – skin, nervous system
 Investigated cleavage
 Discovered chick notochord and mammalian egg
Caspar Friedrich Wolff
 Embryo development occurs through progressive remodeling and
growth
 Supported the theory of epigenesist
Matthias Schleiden & Theodore Schwann
 Schleiden – botanist
 Schwann – zoologist
 Along with Rudolf Virchow, made discoveries that led to the
development of Cell Theory
1. Cells are the basic structural and functional units of life
2. Cells come only from preexisting cells (refuted
abiogenesis)
3. Cells contain genetic information which is inherited by
offspring through genes
 With the development of cell theory, it was proven that development
must occur via epigenesis
 Foundation of modern embryology was laid down, embryology
became a bona fide science
August Weismann
 Studied sea urchin egg fertilization & cleavage
 Developed the germ plasm theory
 Theorized that development follows a mosaic pattern
 Germ Plasm Theory
o Distinguishing somatic cells from germ cells
o Somatic cells are simply vehicles for protecting and
perpetuating the germ plasm
o Germ cells are the ones chiefly involved in the
perpetuation of species
o Germ cells five rise to both somatic and germ cells, but
inheritance is through germ cells only
o Mutations in somatic cells (as in the first generation) is
not inherited by offspring since it does not concern the
germ cells
o Mutations in germ cells (as in the second generation9 is
inherited by offspring and is reflected in both somatic and
germ cells
 1st Generation 2nd Generation 3rd Generation
Soma
Germ Cells
 Mosaic development
o Weismann’s Theory of Nuclear Determiantion
o Zygote contains 2 nuclei derived from both sperm and
egg
o Nuclei contains factors that are asymmetrically distributed
in daughter cells
o Fate of each cell predetermined in egg by factors it would
receive during cleavage
o 4 blastomeres w/ nucleus with different morphogens
o Mosaic model since egg is considered a mosaic of
discrete localized determinants, thus mosaic
development
Ernst Haeckel
 Recapitulation Theory/Biogenetic Law
o An animal in its individual development passes through a
series of stages like those in the evolutionary
development of the race to which it belongs
o Ontogeny recapitulates phylogent
 Embryonic development of an individual (ontongeny) parallels or
summarizes its species’ entire evolutionary development
(phylogeny)
Wilhelm Roux
 German Embryologist
 Performed the Ablation Experiment or Embryonic Extirpation
o At the2-cell stage, destroyed one of 12 blastomeres with
hot needle and allowed embryo to develop, determining
whether the remaining cell will give rise to only a half
embryo or could restore the deficiency during subsequent
development
o Only half embryo was formed
 Proved Weismann’s mosaic development, wherein character and
fate of each cell is determined at cleavage
Hans Adolf Eduard Driesch
 It is possible to remove large pieces from taking away some and
thus interfere in many ways, yet not affecting the resulting embryo
 Any cell monad in the original cell was capable of forming any part of
the completed embryo
 Led to the terms totipotent (total potential) and pluripotent cell
o Totipotent cells can give rise to all cell types (can produce
normal and complete embryo
o Pluripotent cells can only give rise to some cell types
 Regulative development
o Ability of the embryo to develop normally even if some
cells are removed or rearranged
o Early embryonic cells are totipotent
o In the experiment involving sea urchins, surviving cell
developed into a small (With altered orientation of inner
organs) but otherwise normal larva
 Different size, different arrangement of
organelles
 Totipotent but not highly potent at 4-cell stage
(not perfectly normal)
o This theory created friction between Roux and Driesch,
since accordin to Roux, if one of the two cells dire, the
remains develop into a half embryo
o Note: There is no such thing as pure mosaic or
reregulative development. It depends on particular
phase/time of development
Hans Spemann and Hilde Mangold
 Induction Experiment
o Dorsal lip of blastopore grafted from an unpigmented
species of newt to the blastocoel roof of a pigmented
species
o Second embryo was induced
o This means it can continue its own development and
induce adjoining cells to undergo development too
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 Transplantation Experiment
o Partial 2nd embryo can be induced by grafting a small
region of a newt embryo onto a new site on another
embryo
o Grafted tissue taken from dorsal lip of blastopore was
called the evocator since it was responsible for controlling
organization of complete embryonic body
o Triggers surrounding cells to participate in organ
formation

 Received Nobel Prize in Physiology or Medicine in 1935
Wilhelm Johannsen
 Danish biologist
 Distinguished phenotype vs. genotype
o Genotype – genetic information or endowment of an
organism acquired from its parents
o Phenotype is the visible appearance, internal structure or
biochemistry at any stage of development
 Relationship between genotype and phenotype
o How is the genetic endowment translated/expressed
during development to give rise to a functioning
organism?
o Genotype  translation/expression of genes 
phenotype
o Genes can be manifested only when they are activated
depending on time and space
P. Salome Gluecksohn-Waelsch & Conrad Waddington
 British biologists
 Worked on mutant genes affecting posterior part of embryo
development in mouse and Drosophilia
 Genes encode proteins, which determine the properties of a cell
after development
 Integrated embryology and genetics into a unified science called
developmental genetics
 Epigenetics – mechanism that accounts for the development of an
embryo, a combination of genetics and epigenesis
o Development is brought about by a series of causal
interactions between the various parts
 Interaction of cells and tissues arising from the
movements/activity of the proteins encoded by
genes
 Involves the activation of some genes and the
inhibition of others
o Reminds on that genetic factors are among the most
important determinants of development
 Gene regulation modulates development
 Mutations in genes affects embryonic
development
o Other factors or exogenous factors include radiation,
viruses, etc.
Guidelines
 Preformed
- directive influence that determines the course of normal
development
- happens even before the actual embryogenesis
- depends on the time when it will become active
 Maternal Genes and Morphogens/Molecules found in the Egg
Cytoplasm
o direct initial developmental stages
o cytoplasmic factors/determinants are initially very active in
development
e.g. maternal mitochondria
 Zygotic Genes
o direct development after cleavage
 Progressively Formed
- products of the genes: proteins
- include the environmental factors which affect development along
the way
- has overlap with zygotic genes
- Environmental Factors: Anything outside the womb
e.g. drugs, pollutants, emotional state of the mother
Genomic Equivalence
Genomic Equivalence
- each cell in the body has the same genetic material
- all the information necessary to produce a complete organism is
present in every cell in the organism
Evidences:
1 Freddy = amphibian cloning (Rana pipiens)
2 Dolly = mammalian cloning
 Amphibian Cloning (refer to fig. 1)
o removal of frog oocyte nucleus (haploid) and insertion of adult
somatic cell nucleus (diploid)
o for experiment to be successful, nucleus from grafted tissue/cells
should be derived from embryo at early stages of development (from
cleavage to late gastrula)
o Restriction of Nuclear Potency: the ultimate test of whether the
nucleus of a differentiated cell has undergone any irreversible
functional restriction is to have that nucleus generate every other type
of differentiated cell in the body
 Possible because of genomic equivalence
o Totipotency
 total capacity of a cell to give rise to a complete embryo
 direct the entire development of the organism
 true for cells in the early stages of development
 o Pluripotency Cell Differentiation
 ability to give rise to several but not all types of cells
 gives rise to an incomplete embryo Cell Differentiation (refer to fig. 4)
- generation of cellular diversity
- the process by which cells become different in structure and function
- morphological and functional expression of a portion of a genome
- the different structures of cells give a glimpse of their function
- a cell specialized in one or few synthetic pathways
Cells Protein Synthesized Fxn
erythrocytes (has biconcave shape hemoglobin (from globin O2 carrier
and became enucleated to increase genes)
surface area for more efficient
attachment of haemoglobin; loss of
nucleus shortens life span)
muscle cells (striated, spindle- actin & myosin movement
shaped to accommodate tension)
epidermal cells keratin protection
lens cells crystallin sight
plasma cells (from B-lymphocytes) immunoglobulins immunity
neurons neurotransmitters (eg.
Acetylcholine,GABA)
pigment cells melanin

Fig 1. Amphibian Cloning

 Mammalian Cloning (refer to fig. 3)
o mammary gland cell nucleus fused with an enucleated oocyte, then
implanted in a surrogate mother (different breed of sheep)
o of the 434 sheep oocytes, one survived and gave rise to Dolly
o proves that the nuclei of adult somatic cells can be totipotent and
no genes necessary for development have been lost or mutated in
a way that would make them non-functional
o Dolly: experienced premature aging throughout her lifetime (refer Fig. 4 Cell Differentiation from Zygote to 3 Germ Layers
to fig. 2)
- the biological clock of the original adult cell used was not reset Housekeeping Genes VS Luxury Genes
in the cloning process o Housekeeping Genes: genes that are always expressed in all cells
- although the organism is “young”, the cells comprising it are and needed for vital function of cells
“old” o Luxury Genes: genes that are expressed only in specialized cells;
allows differentiation of cells in function and structure; allows the cell
to be more efficient.

In the following sets of gene products in two cell types:

Cell types A & B share a common set of housekeeping gene products and a set
of luxury gene products that represent the A or B developmental program

e.g. If cell A was an erythrocyte and cell B was a plasma cell:

A A&B B
luxury protein housekeeping protein luxury protein
hemoglobin respiration/metabolic antibodies
enzymes

Selective Gene Expression/Differential Gene Expression

Fig. 2 Dolly and her mother
- Expression of certain genes occur only in certain cells at certain
Fig. 3 Mammalian Cloning periods
- Genes are activated differently in phase of development, kind of cell
- Derepression of genes in differentiated cells: Most genes are
repressed in cells
- Time (phase of development) & space (specific cell)
 Selective Gene Amplification Morphogenesis and Pattern Formation

Selective Gene Amplification Morphogenesis: a set of processes that mold the internal and external
- A certain gene is activated and its products are amplified. configuration of an embryo
- An increase in the number of certain genes even in the absence of
mitosis. Pattern Formation: spatial and temporal distribution or organization of
- Important: adaptive response for meeting the synthetic requirements differentiated cells; takes place under tight genetic control
for the developing egg e.g. development of the arms (upper, lower, and fingers)
- One of the mechanisms that occur before cleavage
- Genes are often the lampbrush chromosomes Pattern Formation Morphogenesis
the laying down of the the realization of the morphogenetic
Lampbrush Chromosomes morphogenetic blueprint or body blueprint or body plan
- Maternal genes in the oocyte undergo selective gene amplification; plan
these genes are the “lampbrush” chromosomes example: examples:
- External manifestation of the highly activated genes; the product of 1. defining the main body axes 1. limb formation
which are needed in the early phase of development of the embryo 2. establishment of the fundamental
o Start of cleavage: zygotic genes are not active yet there axes
is rapid cell division. 3. branching of ducts within glands
o The great demand for genes during the early stages of 4. formation of the loops and whorls
development must be met for normal course of of fingers (fingertips)
development; if not, development would be delayed or
the embryo would be abnormal Processes Involved in Morphogenesis
- During the diplotene stage (first meiotic arrest), up to 1500 nucleoli - following processes are not necessarily in order
could be seen at the periphery of the nuclear membrane - some processes may occur simultaneously
o Highly active cell in the synthesis of ribosomal RNA and
mRNA 1. Cell Proliferation
o characterized by rapid cell divisions which results in the increase in
Embryonic Induction number of cells
o achieved through mitotic division
Embryonic Induction
- one of the most important varieties of embryonic signal calling
- cells induce or influence adjacent cells, changing the cells’ behaviour

Induce change

- responding tissue must possess competence* to be able to respond

to the inducer
*For tissue to be competent, there must be compatible receptor cells.
Fig. 6 Mitosis
2. Cell Migration
e.g. Induction of Neurulation and Axis Development by Notochord or o a process wherein cells or a group of cells move from one part of the
Chordamesoderm (refer to fig. 5)
embryo to another
- in the experiment carried out by Otto Mangold, he obtained different o may involve short migration of individual cells or massive dislocation
regions of chordamesoderm after removing the overlying neural of groups or sheets of cells over a relatively great distance
plate from early neurulae and inserted the chordamesoderm pieces
o During gastrualtion
into blastocoels of early gastrula e.g. invagination and involution
Observations: 3. Cell Aggregation or Cell Adhesion
o if anterior chordamesoderm was inserted, the recipient formed a
o sorting of different kinds of cells according to their future/target
secondary head with balancers, forebrain, and eyes
function
o posterior chordamesoderm inserted in the same way induced a o cells of the same function aggregate together
secondary trunk and tail
4. Secretion of Extracellular Substances
o molecules in intercellular space bind cells
5. Change in Cell Shape
6. Apoptosis
o localized programmed cell death
o normal physiological death, as compared to necrosis (premature
death of cells caused by external factors such as disease)
o apoptosis is important for normal embryonic development
o it is responsible for the following:
 resorption of tadpole tail
 separation of the digits in the embryonic hand and feet
 selective cell death of neurons that leave specialized cell
connections

Fig. 5 Induction of Neurulation and Axis Development by Chordamesoderm

  Antennapedia
o a Hox gene first discovered in Drosophila which controls
the placement of legs
o a loss-of-function mutation in the regulatory region of a
gene converts the second pair of legs into ectopic
antennae
o a gain-of-function mutation convert the antennae into
ectopic legs

Fig. 7 Apoptosis
Morphogenesis and Pattern Formation
Homeotic Genes
o set of genes that specify the antero-posterior axis and segment
identity during the early stages of metazoan development
o critical for the proper placement of certain embryonic structures like
legs, antennae, and eyes
Homeobox Genes
o sequence of 180 base pairs which defines the homeotic genes
o the homeobox codes for the 61 amino acid protein known as the
homeodomain (responsible for proper positioning and binding to
DNA)
o many of the homeobox genes found in Drosophila melanogaster has
also been found in vertebrates (homologous genes)
- called homologous genes since there is a 1:1
correspondence between invertebrate and vertebrate
homeobox genes
o these genes are expressed in highly specified sites and stages of
development
 Bithorax Complex
o a Hox gene first discovered in Drosophila which controls
the differentiation of abdominal and posterior thoracic
segments
o mutation causes the development of a duplicated thorax
Fig. 9 Hox gene mutations
Fertilization
Definition
 Egg + sperm = zygote
 Reaction of three bodies: sperm, egg and secretions
Key Functions of Fertilization
 Combine two haploid cells (egg and sperm) to form a diploid
individual
 Contact of sperm with egg “initiates the onset of metabolic reactions
within the egg that trigger the onset of embryonic development”
Types
 External: gametes are released outside of the female body;
commonly found in invertebrates and in some vertebrates like fishes
and frogs
 Internal: sperms deposited into the female reproductive tract; found
in urodeles, reptiles, birds and mammals

External Fertilization
 Paired spawning in fishes
o Milt (sperm cell + secretions) + egg = fertilization of egg
o Fertilized eggs float away in the current or sink to the
bottom, where they lie with the substrate
Fig. 8 Homologue genes of Drosophila and mouse embryos o E.g. Characins and cyprinids
Hox Genes  Mass spawning in Coho salmon
o an example of a homeobox gene complex o Change in coloration (becomes red),
o these genes are expressed along the craniocaudal axis of the body o Die shortly after secretion
in the same way as they are arranged in the chromosome
 Amplexus and multiple amplexus
o mammalian Hox genes can be arranged in four clusters: A-D
- within a given cluster, there are 13 subfamilies or
Internal Fertilization
paralogous groups of genes
 Urodeles
o paralogous chromosomes are arranged in strict order along their
o Less risky method employed by the salamanders
respective chromosomes, and transcribed in the same order for the
whereby the male deposits packet of sperms called a
5’ end to the 3’ end
spermatophore onto the ground
- possess colinearity: the linear order is directly correlated
o Parts of a spermatophore include a stalk and a sperm
to the order of the regions they affect as well as the
cap
timing in which they are affected
- due to colinearity, when a Hox gene segment is lost, the  Reptiles
o Most reptiles produce sexually
segment develops into a more anterior one while a
mutation that leads to a gain of function causes a o Males depositing sperm within the femals’ genital tract
segment to develop into a more posterior one o Egg cell reaches the outside environment surrounded by
the shell of the egg
  Birds
o Cloacal kiss: used by most birds; male and female touch
their cloaca for a few seconds – enough time to transfer
the sperm from male to female
o Male gets on female’s back and twists his tail under the
female
o Mating behavior of chickens:
 Male makes advances, which the female could
ignore or respond by crouching
 Male mounts female and female everts cloaca
 Male then everts it cloaca, and their vents
meet
o Parthenogenesis may also occur, but rarely in nature
(can be done via selective breeding)
 Mammals
o Exclusively internal, relies on copulation
o Mating behaviors of whales
 Gray whales: a “third party” props the female
against the mating male // #3rdwhale
 Humpback whales: pairs surface vertically
face-to-face
o Structure of the gametes: “Successful fertilization
requires that both gametes undergo maturation”
 Sperm must deliver its half of the genome
 Egg must prevent polyspermy (more than one
sperm has fertilized the egg; zygote usually
inviable), and start the development of the
new organism
Hemipenis
 Male copulatory organ
 Usually held inverted within the body
 Everted for reproduction (via erectile tissue; when engorged with
blood)
 Found lateral to the cloaca
 Surgaces often have folds, papillae or spines (used in anchoring
the male within the female)
How Normal Fertilization and Parthenogenesis Differ
 Normal Fertilization
o Precursor egg cell divides into four
o Egg is fertilized with sperm
o Baby created with one chromosome from each parent
 Parthenogenesis or Virgin Birth
o Precursor egg cell divides
o Egg doubles and divides its genetic material
o Cells and genome combine with two sets of identical
chromosomes from the mother
GAMETOGENESIS
 No cells are immortal yet sex cells are immortalized upon fusion:
they die and yet they live by passing all their genetic information
Oogenesis
Role: Prevent polyspermy and start development of new organism
Frog oocytes consist of:
 Nucleus / Germinal vesicle
 Animal pole
 Nucleoli
 Around 1500
 Where rRNA is transcribed
 Lampbrush chromosomes - Corresponds to mRNA
 Transitory structures that exist during meiotic arrest at
diplotene stage of meiosis 1
 Present in female gametocytes of most animals except
mammals
 Produce large amounts of mRNA for the developing oocyte
 Plasma membrane
 Vitelline membrane
 Mostly protein fibers and receptors for sperm binding
 Outside PM
 Jelly coat
 Prevents desiccation, mechanical injury and infection
 Allows rapid absorption of water, making the egg larger and
preventing predation
 Cytoplasm (where a lot of yolk is found, hence known as vegetal pole)
 Vitellogenin
 Refers to both the gene and its product
 A glyco-lipo-protein which is a precursor to egg yolk in female
insects, amphibians, reptiles, birds and platypi
 Mainly made up of lipovitellin and phosvitin and other
morphogens
 Synthesized in liver, delivered to ovary
 Mitochondria – Balbiani Bodies
Mammalian follicular growth
 45 years or so and starts prior to birth (oogonia is undergoing mitosis)
 Consists of oogonia (now oocyte) and its adjacent follicle or nurse cells
Pre-antral Phase – hormone independent development
 Primordial follicle
 Originally 7 to 2 million at birth
 Found at the cortex
 Contain immature oocytes surrounded by flat granulosa cells
 Quiescent; shows little to no biological activity
 Arrested at prophase 1 (diplotene) – before puberty
 Recruitment: process by which primordial cells “wake up”
 Eccentrically located nucleus
 Stromal cells: connective tissue surround the developing follicle
 Stock piles on nutrition
 Primary/Unilaminar follicle
 Granulosa cells change from flat to cuboidal
 Oocyte genome is activated
 Zona pellucida starts to form around oocyte, separating it from
granulosa cells
 Secondary/Multilaminar follicle
 The moment zona pellucida appears, follicle is known as
secondary follicle
 Second layer of granulosa cells is formed
 Theca cells surround the basal lamina and become the theca
interna and theca externa
 Formation of a network of capillaries
 Well-formed zona pellucida present
 Non-cellular
 Glycoprotein
 Prevents polyspermy
 Prevents premature egg release
 Prevents tubal or ectopic pregnancy
Antral Phase – hormone-dependent development
 Antral follicle
 The antrum, a fluid-filled cavity next to the oocyte, starts to
form from fusion of intercellular spaces
 Antrum is the result of coalescence of intercellular spaces
 Components in order of increasing distance from oocyte
 fully grown oocyte
 zona pellucida
 prevents premature release of the egg
 barrier to sperm entry
  around 9 layers of granulosa cells (membrane
granulosa surrounds antrum directly)
 basal lamina/basement membrane
 theca interna (immediately surrounding granulosa)
 capillary net
 theca externa (connective tissue)
 Other cells (granulosa and theca cells) continue to increase via
mitosis while antrum volume increases
Figure 1: Stages of Oogenesis
 Mature/Graafian follicle
o Well-formed antrum is present and it takes up most of the
egg’s volume
o Granulosa cells start to differentiate:
 Corona 9embran: attached to zona pellucida;
supplies vital proteins to cell
 Membrana: interior to basal lamina
 Periantral: adjacent to the antrum
 Cumulus oophorus: connects 9embrane and corona
9embran; anchors the ovum to the 9embrane
granulosa
 A relatively low number of mature follicles may
cause or indicate fertility
 Egg is ready to be released/ ovulated
 Corona radiata
 Cells are seemingly interdigitating
 Due to presence of microvilli on apical surface
Spermatogenesis
 Ultimate goal: to deliver the other half of the genome
 Spermiogenesis – trend: gets smaller and smaller
 Process by which developing sperm mature into motile
spermatozoa
 Sperm are 50-70 micrometers long and are ovoid-shaped
 About 200 million sperms are ejaculated into the human uterus
prior to fertilization
 Cytoplasmic bridges link groups/cluster of maturing sperm so
that their maturation is synchronized
 Sertoli cells
 “Nurse” cells for the developing sperm cells
 Control the entry and exit of hormones, nutrients,
and other chemicals
 Phagocytize residual bodies
 Their junctions form the blood-testis barrier
 Sperm structure – streamlined to allow efficient locomotion
 Head: contains the genetic material in its nucleus,
and the acrosome, which contains the hydrolytic
enzymes needed for penetration into the egg
 Some enzymes in the acrosome:
hyaluronidase, acrosin (a protease
used to create a hole in zona pellucida),
galactosyl, transferase, and glucose
amyladase
 Neck: contains mitochondria which provide ATP
needed by flagellum on its way to the egg, in spiral
configuraiton
 Tail/Flagellum: for locomotion and rapid movement
Figure 2: Stages of Spermatogenesis
 Components
 Type A1 spermatogonia: undergo mitosis and give
rise to types A2, A3, A4 spermatogonia or light and
dark
 Type B spermatogonia: undergo meiosis
 Primary spermatocyte
 Secondary spermatocyte
 Spermatids
 stll joined by cytoplasmic bridges that
allow synchronous development
 undergoes spermiogenesis to form
spermatozoa
 Residual bodies – remains of cytoplasm after
spermiogenesis
 Capacitation
 Process by which sperm are “primed” for contact
with the egg
 Functional maturation occurs in the epididymis
 Involves the alteration of molecules on the surface
of the sperm and the increase in sperm motility
 Occurs inside the female reproductive tract and
takes up to 7 hours
 Glycosyltransferase is unmasked on the sperm
plasma membrane and prevents premature release
of enzyme
 Only a few hundred sperm can reach the egg due to
presence of WBC and acidic conditions
Mammalian Fertilization
 Mammalian Fertilization
o Spermatozoa penetrate the cells of cumulus oophorus and
corona radiata
 Via hyaluronidase enzyme (found in acrosome)
which dissolves the extracellular matrix around the
cells of the corona radiata
 CO and Corona radiata cells have to be dispersed
  Strongly aided by the swimming movement of the
sperm
 Follicle cells act as guard
o Zona Pellucida + Sperm
 Zona pellucida allows only sperm of the same
species
 ZP3: primary receptors
 N-acetylglucosamine
 Galactose
 Zona Receptor Kinase
 ZP2: maintains binding of the sperm (inner
acrosomal membrane)
 ZP1: cross link the ZP3 and ZP2
 Sperm adhesion proteins
 Galactosyltransferase
 SP 56
 P95
 Process: ZP3 receptors --> cascade of transduction
pathways --> G-protein and kinase --> IP3 synthesis
and release of calcium ions from ER --> Calcium
ions release other acrosomal proteases
o Acrosome Reaction
 Localized fusion of the outer acrosomal membrane
and the plasma membrane of the spermatozoon
 This leads to the opening of ion channels in
the egg plasma membrane, allowing the entry
of sodium ions.
 The entry of sodium ions causes the
depolarization of the membrane.
 *Fast Block to Polyspermy /Transient
 another term used for depolarization
(change in charges), which occurs
within about 1 to 3 seconds after a
sperm binds to the vitelline membrane
 prevents more than one sperm from
fusing with the egg
 Release of soluble enzymes from acrosome. The
acrosome acts like an enzymatic drill that allows the
passage of the sperm into the egg cytoplasm.
 The tip of the acrosomal process is coated
with a protein that adheres only to specific
receptors on the egg vitelline membrane.
 “Lock-and-key” system of molecule
recognition ensures that only sperms of the
same species will fertilize the egg.
o Fusion of Sperm and Egg Plasma Membrane
 Due to Fertilin (from sperm) and Integrin (from
egg)
 Point of primary fusion called equatorial domain is
parallel to the sperm PM
o Cortical Reaction
 Refers to the changes that happen in the cortex
of the egg cytoplasm
 Cortical granules are exocytosed and release
proteases that cleave the proteins that link the
vitelline envelope to the cell membrane
 Mucopolysaccharides also released by the granules
make a concentration gradient, causing water to
flood in and swell the space between the vitelline
envelope and the cell membrane
 Other enzymes released from the granules harden
the vitelline membrane (now called the fertilization
envelope after separation from the oocyte) and
 Other Features of Mammalian Fertilization
release sperm bound to it (in the case of sea
urchins)
 Egg is fertilized with sperm. Fusion of the egg and
sperm triggers the release of Ca+ ions into the egg
cytosol.
 In response of the increase in Ca+
concentration, the cortical granules in the
egg fuse with the plasma membrane
 The cortical granules release their
contents...
- Enzymes – separate the vitelline
membrane from the plasma membrane
- Mucopolysaccharides – draws water
into the perivitelline space, pushing the
vitelline membrane further away
- Enzymes, then, harden the vitelline
membrane, which now becomes the
fertilization envelope.
 Baby created with one chromosome from each
parent
o Slow Block to Polyspermy
 Slow but permanent block to the entry of other
sperms
 The two blocks account for the Principle of Double
Assurance or Synergistic Mechanism
 CG + PM  exocytosis of cortical granule
molecules
 Cortical granules + plasma membrane promotes
exocytosis. Process:
i. GAGs (glycoaminoglycans) create
osmotic gradient. Water enters. Space
between plasma membrane and vitelline
membrane swells.  perivitelline space
+ fertilization membrane formed
ii. Proteases cleave protein links between PM
and VM which become the same thing.
iii. Peroxidase promotes hardening of zona
pellucida and inactivates receptors,
leaving runner-up sperms out in the cold.
o Amphimixis
 Fusion of pronuclei – enlarged, with diffusion of
DNA
 Before the sperm enters, the egg is arrested at
metaphase II. Meiotic division then resumes as the
sperm enters, resulting to release of 2nd polar
bodies
 Sperm enters completely
 DNA in sperm, bound mostly by protamine instead
of histones because protamine is used for more
intense condensation, starts undergoing
decondensation, and threadlike chromosomes start
to appear. Head is now male pronucleus.
 The egg is also decondensed and forms the female
pronucleus
 Nuclear membranes of both pronuclei fuse into one
and then disintegrate. Fusion of pronuclei then
occurs.
 Chromosomes are recondensed (by histone
replacement) in mitotic spindle.
 Therefore, a diploid nucleus has already appeared
in the 2-cell stage, preceding the zygote. Both
specialized cells “die,” yet they live on in the zygote.
 Sperms provide genes and centriole only
 Microvilli around the egg take the whole sperm into the
egg
  The sperm flagellum divides, forming two centrosomes
 These centrosomes are used for cell division;
unfertilized mammalian eggs have no
centrosomes.

Establishment of Polarity

 For amphibians:
 Before fertilization, primary polarity (presence of animal and
vegetal pole) is already present in the amphibian egg.
 Rotation of cortical cytoplasm with respect to internal cytoplasm
(around 30 degrees) occurs due to sperm entry
 The point opposite sperm entry becomes the gray crescent – an
area of reduced pigmentation
 Gray crescent defines the dorsoventral and craniocaudal /
anteroposterior axes and with these two established, the
mediolateral axis can be geometrically determined – secondary
polarity is established.
 Gray crescent is on the dorsal and posterior half of the embryo.
 The gray crescent is also the site where gastrulation begins.
Figure 1: Cleavage in different organisms

Cleavage  Types of eggs

o Based on amount of yolk:
 Definition  Macrolecithal: large amount of yolk; amount of yolk
o A series of rapid cell divisions is greater than the cytoplasm
o Blastula: hollow ball of cells  Teleosts, birds, monotremates,
o Blastocoel: cavity inside a blastula gymnophiona (legless amphibians)
o Morula: solid ball of cells  Microlecithal: small amount of yolk; amount of yolk
o In a somatic cycle: is much less than the cytoplasm
 G1: 0 hours – 5 hours  Some embryologists describe
 S: 5 hours – 12 hours microlecithal eggs as alecithal (no yolk)
 G2 : 12 hours – 15 hours or oligolecithal/miolecithal (little yolk)
 M: 15 hours – 16 hours  Amphioxus, marsupials, therian
o In a cleavage cycle (30 minutes), there are no G1 and G2 mammals
phases, hence there is no cell growth  Eggs of mammals contain very little yolk
that they are sometimes called alecithal
 Mesolecithal: moderate amount of yolk
 Lampreys and amphibians
 Alecithal: no yolk present

o Based on distribution of yolk:

 Telolecithal: concentrated in one region of the egg
(reptiles and birds, macrolecithal and mesolecithal
eggs)
  Isolecithal: equally distributed throughout the egg  Second cleavage: perpendicular to the first
(microlecithal eggs)  Third cleavage: radial and synchronous
 Centrolecithal: concentrated in the center of the  Fourth cleavage: circumferential and asynchronous
egg (insects, macrolecithal egss)  Fifth cleavage: centrifugally

Notes: Continuous cleavages divide the blastoderm into a mass of 5 to 6 layers

thick until they reach 60,000 or so; Cells are linked together by tight junctions
(zonula occludens)

Figure 2: Types of Eggs and Cleavage Patterns
Notes: The blastoderm contains about 60, 000 cells by the time the egg has
been laid
Cleavage Patterns
Embryo forms as a cap of cells sitting on top of the massive yolk
Marginal zone in between area pellucida and opaca
 Holoblastic: complete cleavage
 Isolecithal egg: sparce, evenly distributed yolk
 Radial cleavage: echinoderms, amphioxus
o Displaced radial cleavage: division of the
egg is complete but cells are unequal in size;
occurs in mesolecithal eggs
 Spiral cleavage: annelids, molluscs and flatworms
 Bilateral cleavage: tunicates
 Rotational cleavage: mammals and nematodes;
with no obvious polarity
 Cleavage furrow passes through the entire egg
 Equal (resulting cells have the same amount of yolk; in
microlecithal eggs) or Unequal (some resulting cells contain
more yolk than the others; in mesolecithal eggs)
 Results in the formation of blastomeres surrounding
blastocoels
 Meroblastic: incomplete cleavage (only in area with living cytoplasm,
other parts are inert)
 Telolecithal egg: dense yolk throughout most of cell
 Bilateral cleavage: cephalopods, molluscs
 Discoidal cleavage: fish, reptiles and birds
(division of egg is confined to disc of cytoplasm)
 Cleavage happens only in a disk at the animal pole
 Results in the formation of a blastodisk atop the yolk
 In macrolecithal eggs
Avian Cleavage
 Discoidal cleavage: division of the egg is confined to a disc of
cytoplasm (occurs in strongly telolecithal eggs due to their yolk)
 Discoidal cleavage in Avians
 Meroblastic, macrolecithal
 Bulk of oocyte consists of yolk
 Cleavage occurs in the blastodisc (a small region of active
cytoplasm at the animal pole of the egg)
 Cleavage does not extend in yolky cytoplasm
 Bases of early cleavage cells are open to the underlying yolk
 Direction of cleavage
 First cleavage: furrow appears centrally in blastodisc to
produce a single layered blastoderm (meridionally)
 Subgerminal cavity
 The space between the blastoderm and the yolk
 Appears when embryo is about 100 cells thick
 Produced when blastoderm cells absorb fluid from the albumin
and secretes it between themselves and the yolk
 Albumin – more fluid than yolk
 Area pellucida
 Clear central region, one cell thick
 Produced when deep cells in the center of the blastoderm are
shed
 Shed cells appear to die
 Area opaca
 Peripheral ring of blastoderm cells that are not shed
 Avian blastoderm
 Blastoderm: disc of cells sitting on top of the yolk
 Dorsoventral axis depends upon the dorsal side
forming away from the yolk while the ventral side is
next to it.
 Initially radially symmetric after egg is laid
 Dense cells form at the posterior marginal zone
(PMZ)
 Primitive streak: develops from the posterior
marginal zone
 Avian embryos
 Due to the presence of highly dense and inert yolk in the eggs,
cleavage is either slowed or blocked
 Complete division is restricted to the least yolky region of the
egg
Mammalian Cleavage
 Holoblastic, equal, not much yolk, no yolk to be seen
 First division
 Extremely slow cleavage (24-36 hours)
 Normally meridional
 Second cleavage
 Asynchronous in the two blastomeres resulting in
crosswise arrangement of the resulting blastomeres
 Unique orientation of mammalian blastomeres in
relation to one another
 One cell divides vertically/meridionally, the other
equatorially  4 blastomeres (cross wise)
 Cortical Rotational: 90 degree shift of the mitotic
spindle in one blastomere
 Early 8-cell stage
 Cells are arranged loosely
 Mid 8-cell stage
 Compaction: cells become flattened, compacted
and tightly joined to allow clustering of the cells
 Promotes adhesion of cells (promoted by E-
cadherin), causing cells to form a compact ball
 Stabilized by tight junctions (zonula occludens)
and laminin
  Blastocyst:
 Inner cell mass and trophoblast
 Morula stage: flattening of cells maximize contact between
cells
 Morula is made up of 16 blastomeres
 Cells undergo cavitation
 Trophoblast cells secrete fluid into morula to create
a blastocoel (cavity)
 Gap junctions (zonula nexus) allow ions and molecules to flow
between cells
 In heart – as intercalated discs  allows
synchronous beating)
 Rapid communication between adjacent cells
undergoing development
 Tight junctions prevent exchange of fluid between embryo and
environment to maintain the integrity of the cells
 Microvilli are also present as another source of cell adhesion
 Fluid secretion in the morula (fluid filled cavity)
 The inner cell mass is positioned on one of the ring of
trophoblast cells
 Trophoblast gives rise to the chorion
 Secretes hormones that regulate the mother’s
immune system which prevents rejection of the
embryo --- embryo can be considered foreign
because it is partly composed of materials from the
father, and is therefore not completely identical to
the mother
 Inner cell mass gives rise to
 Embryo proper: embryo at the end of cleavage is
called a blastocyst (comparable to a blastula in
amphibians and blastodisc in birds)
 Extra embryonic membranes amnion, allantois
and yolk sac
 Also secretes proteins that influences the
division of the trophoblast
 Differentiation of embryos can wait until after the attachment or
implantation of the blastocyst into the wall of the uterus
 After the 6th or 7th day, the embryo begins to bury
itself into the endometrium
 Mammal: Blastocyst :: Bird: Blastodisc :: Frog:Blastula
Note: Evagination – outward folding, vs. Invagination – inward folding
Totipotent – capacity to produce complete embryo, vs. Equipotent – involves
genomic equivalence, each cell will have similar genetic makeup as other cells
Summary:
 2-cell – meridional
 4-cell – meridional and equatorial
 8-cell – compaction
 Morula – caviation
 Blastocyst – differentiation
Escape from the Zona Pellucida
 Blastocyst contained in the ZP expands via Na+/K+ ATPase pump of the
trophoblast; draws water osmotically into the blastocoels
 Embryo enlarges due to increase in number of cells; action of the
Na+/K+ - ATPase pump
 Zona pellucida: prevents premature interaction and adhesion of the
blastocyst with walls of the oviduct
o Otherwise, ectopic pregnancy would occur.
 Mice: protease called strypsin lyses a hole in the ZP; compare to the
bird’s egg-tooth
 Endometrium: catches the released blastocyst on an extracellular
matrix
o ECM: contains collagen, laminin, fibronectin, hyaluronic acid,
heparin sulphate receptors
 Trophoblast cells contain integrins that will bind into the uterine
collagen, fibronectin and laminin; only released at the time that it is
most needed
 Trophoblast cells also synthesize heparin sulphate proteoglycan
precisely prior to implantation
 Once in contact with the endometrium, the trophoblast secretes another
set of proteases, which digest endometrial lining; allows blastocyst to
bury itself into the wall
o Collagenase
o Stromelysin
o Plasminogen activators
 Blastocyst: initially asynchronous division
 Summary of stages:
o Fertilization
o Cleavage
o Attachment
o Hatching (creation of hole in ZP) and Implantation
 Doushantuo Formation in South China: 600M year old fossilized
embryo
Amphibian Cleavage
 Cleavage Pattern: displaced radial cleavage
 Amount of yolk: Mesolecithal (moderate vegetal yolk disposition)
 Holoblastic complete cleavage: Complete division of the egg
 Cleavage period and blastula formation -> rapid, complete at 24 hours
 Sequence of Amphibian Cleavage:
o 1st division: meridional; starts at the animal pole; slowly
extends down the vegetal pole and bisects the grey crescent
o 2nd division: meridional; begins at the animal pole as well;
plane perpendicular to the 1st cleavage
o 3rd division: equatorial; horizontal cleavage plane passing
nearer to the animal pole resulting to four smaller blastomeres
at the animal hemisphere and four larger blastomeres at the
vegetal hemisphere; becomes unidentical with respect to size
due to pressure of yolk
o 4th division: double meridional axis of cleavage perpendicular
to each other
 Axolotl salamander: cleavage furrow extends
through the animal hemisphere
o 5th division: double equatorial; above and below the 3rd
 Unequal holoblastic cleavage: establishes two majpr embryonic regions
o Micromere: rapidly dividing near the animal pole
o Macromere: slowly dividing near the vegetal pole
 Blastocoel is found in the animal pole and its shape is affected by yolk
 Chick: Slit :: Frog: Hemisphere :: Amphioxus : Sphere
 As cleavage progresses:
o Animal region: becomes packed with numerous small cells
o Vegetal region: contains a relatively smaller amount of larger
and yolk-laden macromeres
 Amphibian embryo
o 16 to 64 cell stage: Morula (pl. Morulae; from the Latin
“mulberry”, whose shape it vaguely resembles); solid ball of
cells
o 128 cell stage: blastcoel becomes apparent, and the embryo
is now considered a blastula
Blastulation
Formation of the Amphibian Blastocoel
 First cleavage furrow: small cleft; later develops into the blastocoels;
widens in the animal hemisphere to create a small intercellular cavity
 8-cell stage: small blastocoel at the junction of the cleavage furrows
 Result of expansion of cleavage furrow
 Importance: permits cell migration during gastrulation and prevents the
cells beneath it from interacting prematurely with the cells above it
o Because mesodermal tissues are normally formed from those
animal cells that are adjacent to the vegetal endoderm
precursors, it is plausible that the vegetal cells influence the
adjacent cells to differentiate into mesodermal tissues
[induction process]
 o Delamination: Splitting or migration of one sheet into two
sheets (e.g. mammalian and bird hypoblasts)
o Epiboly: The expansion of one cell sheet over other cells
(e.g. ectoderm formation in amphibians, sea urchins and
tunicates)
Note: Gastrulation of any particular organism is an ensemble of several of these
movements

Figure 1. Amphibian Blastulation

 Why are factors needed to achieve target form of cells? Figure 2. Morphogenetic Movements Involved in Gastrulation
o Because of the totipotency of the cells, they need to be
guided Gastrulation in Amphioxus
 Blastocoel appears to prevent contact of vegetal cells destined to
become endoderm with those cells fated to give rise to the skin and the  Invagination of presumptive mesoderm (endomesoderm)
nerves  Inward movement or involution of deep mesodermal cells or Inner
 While these cells are dividing, numerous cell adhesion molecules Marginal Zone Cells
(CAM) keep the blastomeres together o Eventually line the roof of the archenteron
o EP-cadherins: cell adhesion molecules that keep blastomeres  Formation of archenteron displaces blastocoels; yolk plug from
together, found in between adjacent cells endodermal yolky cells future posterior region of the embryo
o Integrins: connect cytoskeleton of the cells to the extracellular  Cell proliferation and elongation of embryo
membrane (ECM)
 Gap junctions or nexus made up of connexons (building blocks of Gastrulation in Frogs
each connexon: 6 connexins)
o Allow communication between adjacent cells via passage of  Starts at marginal zone
ions and molecules  synchronous development
 Blastopore: for marginal cells' inpocketing (blastopore's dorsal lip =
 Flower-like arrangement
Spemann organizer)
 Hexagonal cavity
 Invagination of less yolky cells -> bottle cells -> pharyngeal endoderm
o Occludin – part of zonula occludens
-> foregut (pharynx, esophagus, stomach)
 The amphibian blastula can be divided into 3 main regions:
o Bottle cells: so-called due to necklike region; less yolky cells
o Roof of blastocoel: region around the animal pole (future
that are first to invaginate
ectodermal layer)
 Archenteron displaces blastocoels; yolk plug occludes blastopore
o Yolk mass: region around the vegetal pole including the large
from endoderm
cells in the anterior (future endoderm layer)
o Marginal ring of cells: found in subequatorial region of the  Involuting inner marginal zone cells (IMZC)/ deeply involuting marginal
embryo, including the grey crescent (future embryonic zone cells – enter into blastocoel, forms thin narrow layer of cells
mesoderm)  Mediolateral intercalation: cells from a single layer rearrange to form
a longer and narrower column which results to convergent extension
 Goosecoid & Noggin genes: activated after vegetal shifts dorsally in o roof of archenteron (single layer)
late blastula; for dorsal marginal region's differentiation & migration  Radial Intercalation: cells from 2 layers pack into a single layer;
 Nieuwkoop center: early organizing structure formed after cortical increase SA
reaction  Epiboly of animal cap and non-involuting marginal zone cells ->
ectoderm; vegetal cells -> endoderm
Gastrulation
 Mesodermal induction: for vegetal organization
 Fate of other cells:
“It is not birth, marriage or death but Gastrulation, which is truly the most
o Remaining vegetal cells -> endoderm
important time in your life”
o Blastopore -> slit -> crescentic -> ring -> dorsal, ventral, and
-Lewis Wolpert 1986
lateral lips of the blastopore
o Yolk plug: internalized
 Migration of cells leading to the formation of the 3-germ layered o Blastocoel: obliterated, replaced by archenteron (primitive gut)
embryo, the gastrula
 Completion bridge – between archenteron and displaced blastocoel
 Hihgly orchestrated movement of cells that establish the vertebrate
 Blastopore – opening, lip - cells
body plan
 Utilizes various morphogenetic movements:
o Invagination: infolding of cell sheet into the embryo (e.g.
sea urchin embryo)
o Involution: inturning of cell sheet over the basal surface of the
outer layer (e.g. amphibian mesoderm)
o Ingression: migration of individual cells into the embryo (e.g.
sea urchin mesoderm, Drosophila neuroblasts)
  Epiblast cells  cuboidal/columnar
 Mesenchymal  bottle-shaped
 Formation of the three germ layers
o Head mesenchyme
o Head process (found where hensen’s node is) and
notochord: supports the embryo with the somites at its either
side
o Somites: muscle cells; differentiate to
 Epimere
 Mesomere
 Hypomere (lateral plate mesoderm): differentiates
to somatic and splanchnic mesoderm; gives rise to
the coelom
o Endoderm: lining of the digestive tract
o Ectoderm: skin epidermis
Figure 3. Gastrulation in Amphibians
Areas around Primitive Streak
Gastrulation in Birds  Area pellucida: lucid or clear area surrounding the primitive streak
 Area opaca: darker area next to area pellucida
 Occurs with less processes
 End point: formation of primitive streak (initial structure; crucial for
gastrulation)
 Migrating cells have different shapes (from columnar to
mesenchymal/flask-shaped/bottle-shaped)

Formation of Hypoblast
 Ingression  primary hypoblast (cluster of cells), also called
polyinvagination islands (misnomer)
 Primary hypoblast (from posterior marginal zone) + secondary
hypoblast (definitive hypoblast)
 Condensation of hypoblast cells  primitive streak
 Stages:
o Congregation of hypoblast cells around the Koller’s sickle
(posterior marginal zone cells)
o Hypoblast island cells coalesce to form the primary hypoblast
layer, which meets the endoblast cells and primitive streak
cells at Koller’s sickle
o Secondary hypoblast cells migrate anteriorly
o Primitive streak cells form a layer between the hypoblast and
epiblast (from blastoderm)
o Primitive streak becomes a defined region of the epiblast, with
Figure 4. Gastrulation in Birds
the cells migrating through it (mesoderm and endoderm)

Cell Movements of the Primitive Streak of the Chick Embryo

 3-4 hours (after fertilization): thickening of the blastoderm anterior to
the Koller’s sickle
 7-8 hours: area pellucida enlarges; primitive streak takes shape
 15-16 hours: definitive primitive streak stage; Hensen’s node and
primitive groove are distinct
o Thickening of anterior portion of primitive streak  Hensen’s
node (cluster of cells in the anterior part)
o Primitive pit: minute cavity of Hensen’s node
Formation of Germ Layers
1. Ingression from epiblast via Hensen’s node  pharyngeal
endoderm; germinal crescent  primordial germ cells (PGC); head
mesenchyme, prechordal plate and chordamesoderm  head
process and notochord
2. Lateral migration  endoderm and EEM
3. Epiboly of epiblast  ectoderm
 Spindle-shaped  forms layer  assumes shape of epithelium
 Formation of the primitive streak (3rd – 18th hour)
1. 3rd – 4th hour: condensation of epiblast cell in the mid-dorsal region
2. 7th – 8th hour: elongation of primitive streak towards anterior region
3. 12th – 17th hour: formation of the primitive groove, primitive
ridges, primitive folds and primitive knot (Hensen’s node) in most
anterior region (Primitive pit  found in Hensen’s node)
4. 18th hour: maximum length of primitive streak (definitive)
Gastrulation in Mammals
1. Delamination of inner cell mass of the blastocyst (ICM + trophoblast)
 epiblast and hypoblast
o Hypoblast: first cells to segregate out of ICM; forms only
extraembryonic endoderm -> lining of yolk sac
o Epiblast: remainder of ICM; contains future ectodermal cells;
also, cells that will migrate through primitive streak ->
endoderm and mesoderm
2. Differentiation of trophoblast into cytotrophoblast and
syncytiotrophoblast
o Syncytium: no cytokinesis results in multi-nucleated cells
o Syncytiotrophoblast: multi-nucleated (division without
cytokinesis) big cells; invades uterine lining producing
trophoblastic lacunae
o Cytotrophoblast: nuclei separated by plasma membrane;
active in cell division; proliferates to create chorionic villi; fully-
fledged cell, separated plasma membrane individually
3. Formation of the primitive streak
o Primitive Streak: condensation of epiblast cells in mid-dorsal
region; elongates towards anterior region
4. Formation of the three germ layers
5. Extra-embryonic mesoderm before definitive mesoderm
6. Primary yolk sac replaced by secondary yolk sac
7. Later stages of gastrulation: formation of somites
 o Epimere, mesomere, lateral plate mesoderm (somatic and
splanchnic)

Figure 6. Extraembryonic Membranes

Nidation/Implantation
 Bilaminar: at 14th day (2 weeks)
 Trilaminar: at 21st day (3 weeks)
o Craniocaudal folding with lateral folding
 Placenta: site of oxygen exchange of fetal and maternal blood
Figure 5. Gastrulation in Mammals o Umbilical cord: former connecting stalk; connection between
mother and offspring containing arteries and veins from fetus
Extraembryonic Membranes o Chorion: fetal component; gives rise to blood vessels which
 Amnion connects mother and offspring
o Filled with amniotic fluid o Decidua cells (endometrium): maternal component;
o Innermost sac; for protection from mechanical factors (shock functional cells; start to become enlarged as they accumulate
absorber), desiccation and adhesion glycogen and lipids; allows nourishment of embryo
o Allows for movement of embryo as well (“private swimming  Decidual reaction
pool”) o Trophoblastic lacunae: glandular secretion of mother
o Derived from the ectoderm and parietal mesoderm  Enzymes involved:
 Chorion o Stromelysin
o Outermost sac from cytotrophoblast o Collagenase
o Attached to the mother; has villi (primary, secondary, tertiary)  Embryo (an antigen to the mother’s body due to presence of paternal
to increase the chorionic surface area for nutrition of the rapidly DNA) can attach to mother’s uterus without getting attacked by the
proliferating cells of the embryo and drawing gases and mother’s immune system by secreting interleukin-2 from helper T-cells
nutrients from maternal tissue (endometrium)  Diapause: delayed implantation
o Highly porous  Roe deer: embryo at 30-cell stage release PAG to maternal receptor
o Derived from ectoderm and splanchnic mesoderm
o Respiration for birds: free diffusion of gases
 Allantois
o For excretion
o Becomes part of umbilical cord
o Ventral evagination of splanchnic mesoderm
o Derived from endoderm and splanchnic mesoderm
o May fuse with chorion and form chorioallantoic membrane
o In birds (balut), rich in uric acid which leads to arthritis
 Yolk sac
o From evaginations (outward folding) of hindgut and PMG
o Source of primitive blood cells (hematopoietic organ)
o Primary yolk sac (large) is replaced with secondary yolk sac
(small, definitive and final)
o Morphology is more conservative than physiology
 Presence of yolk sac even without the yolk
 All the nutrients, as well as function of excretion, is
provided by the mother
 Derived from endoderm and splanchnic mesoderm

 Allantois and yolk sac are non-functional in humans because

nutrients are acquired from the mother; they may have other
functions
 In birds, chorion + allantois = chorioallantoic membrane

Figure 7. Implantation of Mammalian Embryo

Summary Intrinsic Factors of Cellular Migration
 Zygote -> Gastrulation via segmentation -> Ectoderm, Mesoderm,
Endoderm
 Ectoderm
o Epidermis
o Central nervous system (neural tube)
o Peripheral nervous system (neural crest cells)
 Mesoderm
o Dermis
o Kidneys
o Reproductive Organs
o Bones
o Muscles  Lamellipodia – cytoskeletal actin projections for initiating movement; much
o Vascular System broader (broad sheet)
 Endoderm  Filipodia – threadlike, cytoplasmic processes that guide/aid movement;
o Intestines much thinner and longer
o Lungs  Extracellular Matrix (ECM) – rich in fibronectin, laminin, type IV collagen,
o Liver and integrin
o Laminin (and Fibronectin) acts as migration pathway
Assignment: o Type IV collagen is a unique type of collagen found in the basal
What mechanisms are involved in gastrulation? Give the structural and lamina, between the epithelium and the connective tissues
molecular bases of gastrulation. o Integrins bind cells with molecules
 Intrinsic factors: cytoplasmic processes (lamellipodia, filipodia)
 Lamellipodia – leading edge will migrate towards target, *Rapid cell division gives rise to
flat Frog: blastula
 Filipodia – thread-like Human: Blastocyst
 Extrinsic factors Chick: Blastoderm
 ECM rich in fibronectin – migration highway
* “Cells in blastula are made up of a mass of cells that are more or less, uh,
 Laminin - endometrium
yung totality of the size is similar to the original zygote because there is
 Collagen type IV – usually found between epithelium & continuous repeated division of cells. Greater in number (of cells ata), but size
CT (ideal for migrating cells) (of each cell) becomes smaller and smaller”
 Integrin – binds to laminin (functions in cell binding)
 E-cadherin, scatter factor *Unique property of cells that make up the blastula: totipotent and equipotent
 Lose cell adhesion molecules – to allow cells to migrate inward (can give rise to all cell types and exhibit genomic equivalence).
 Hyaluronic acid – highly poler (N-acetylglucoseamine, d-
glucoronic acid) * Cells in blastula are totipotent and equipotent, whereas cells of gastrula are
What are the molecules involved in cleavage? qualitative; totality of size is similar to zygote due to continuous division
 Microfilaments – for the cleavage furrow resulting only in increased number of cells
 Tubulin – component of microtubules – bring chromosome to
opposite poles Mass morphogenetic movement (3 to 4 types)
 Cyclin and histone – produced by mother for breakdown of nuclear  Amphibian – involution
membrane  Avian/Mammalian – ingression

*lead to formation of 3 germ layers (blueprint for organogenesis)

Diapause
 Delayed implantation

Fate Maps of Xenopus (South African clawed frog)

 shows prospective locations of future regions/organs in adult frog
 Example:
o Ectoderm (differentiated under the influence of notochord) 
Neural ectoderm  Neural Tube CNS
o Mesoderm (undegoes involution)  somites
(chordamesoderm), blood, kidney, heart
o Endoderm(made up of vegetal/yolky cells)  inner Organogenesis
lining/epithelium of digestive tract, organs derived fom primitive
gut  second most important journey of cells!, includes morphogenesis
 directed by factors such as Spemann organizer (dorsal lip of blastopore) (structuring of the embryo), give rise first to primary organ rudiments
 somehow manifested by the formation of 3 very important vertebrate
* The gastrulation is made possible simply due to the change in shape of cells. structures: notochord, somites, neural tube surrounded by prospective
Also, in the assumed certain shapes they exhibit cytoplasmic processes. epidermis.
 Types of Neurulation

Neurulation  Primary Neurulation

 formation of neural tube (forerunner of the CNS, precursor of brain and o performed by most vertebrates
spinal cord) o anterior portion of the CNS in all vertebrates
 CNS has)longest development (from week 3 to 16, and even after birth or o origin: prospective epidermis
postnatal)) even if structural components (neurons, glial cells, etc.) are o Steps:
already positioned prior to birth. First trimester/3 months= very crucial 1. initial unilaminar, simple cuboidal epithelium
 development of neural tube starts in the first trimester making it a very 2. columnarization or change in cell shape to simple columnar
important period form
3. rolling / folding / bending via invagination of propspective
neural ectoderm
4. closure by the joining and fusion of cells
5. completion of the neural tube
 Secondary Neurulation
o done by fishes and some vertebrates
o posterior region (sacral/tail) of CNS in all vertebrates
o origin: mesenchyme
o Steps:
1. dispersed, loosely arranged mesenchyme (highly
undifferentiated cells)
2. mesenchymal condensation or aggregation
3. formation of solid mass of cells called medullary cord or
neural rod
4. epithelial transition / cavitation by hollowing out of the central
portion
5. completion of neural tube
*Dorsal Surface View:
Neuropore -opening on top of neural tube in Late Neurula stage.

*Sagittal View:
Epidermis also moves to replace invaginated cells/columnar cells that have
undergone folding.

* Neurenteric Canal – temporary canal connecting posterior part of neural tube

and archenteron. Between the notochord and mesoderm: protects overlying
archenteron and blastopore.
*Union of neural folds at the middorsal region: starting point = slightly anterior
part of midbrain.
Chick Neurulation
 appear near the embryo’s dorsal surface. They will give rise to most of the
skeleton’s axial portion, skeletal muscles, and much of the dermis.
 Days 24 to 25 – By now, some embryonic cells have given rise to
pharyngeal arches. These will contribute to the formation of face, neck,
mouth, nasal cavities, larynx, and pharynx.
Closure of Neural Tube

 has multiple sites; closes about the midbrain first, then proceeds craniad
and caudad
 Anencephaly – results from failure to close by the anterior neuropore
 Spina bifida – results from failure to close by the posterior neuropore; also
caused by failure of vertebrae to fuse
 as anterior part is undergoing organogenesis to form primary organ
rudiments, posterior part’s primitive streak is undergoing regression  Test Your Skills!
anterior part more advanced
 Steps:
1. columnariation of neural plate (NP)
2. bending of NP
3. meeting of neural folds (NF)
4. union of NF
5. closure of neural tube (NT)
6. dispersion of neural crest cells

Purse-String Effect - constriction of apical region

 changes in cell shape to produce wedge-shaped neural ectoderm
 involves MT and MF molecules 1 Neural plate, 2 Primitive streak, 3 Primitive nodes, 4 Neural groove, 5
 involves median hinge point cells (MHPC) and dorsolateral hinge point Somites, 6 Cut section of the amnion, 7 Neural folds
cells (DLHPC)
 MHPC – shorter cells closely apposed to the notochord that allows
invagination of neural ectoderm and more bending undergo change in cell
shape
 DLHPC – longer cells accounting for shorter bending; flexibility allows
inward folding and actual fusion of cells towards middorsal region. The
one at the right and left, at the junction bet. Ectoderm, the prospective
epidermis and the neural ectoderm which undergoes invagination.
 folding will not occur until purse-string effect occurs for each individual cell

Mechanisms of Neurulation
 Induction
o Notochord  MHPC to decrease their height and become columnar 1 Neural tube, 2 Neural fold, 3 Neural groove, 4 Somites, 5 Neural crest, 6
then wedge-shaped, acts as anchoring point to allow NP cells to Protrusion of the pericardium, 7 Cranial neuropore, 8 Caudal neuropore
elevate
 Extrinsic Factors
o Ectodermal cells are pushed toward the midline
 Changes in cell shape via
o Microtubules – maintains their columnar shape (through its tubulin
component); displaces parallel to the long axis of the cells
o Microfilaments (actin) – promotes apical constriction; disposed on
the apex of the cell (perpendicular)

Human Neurulation
 Day 15 – A faint band appears around a depression along the axis of the
embryonic disk. This is the primitive streak, and it marks the onset of
gastrulation in vertebrate embryos
 Days 18 to 23 – Organs start to form through cell divisions, cell
migrations, tissue folding, and other events of morphogenesis. Neural
folds will merge to form the neural tube. Somites (bumps of mesoderm)
 Blastocyst

Inner Cell mass Trophoblast

Epiblast Hypoblast Cytotrophoblast

Extraembryonic
Embryonic Epiblast Amniotic Ectoderm Syncytiotrophoblast
Endoderm

Embryonic Ectoderm Primitive streak Yolk Sac Endoderm

Embryonic
Notochordal Process Mesoderm
Endoderm

Extraembryonic
Allantoic Endoderm
Mesoderm

Embryonic
Mesoderm