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SEMBRYOLOGY Its Scope, History, and

Special Fields
Historical Background

Germ-cell line was all-important for


perpetuation of species; soma was
primarily vehicle for protecting and
perpetuating germ plasm

Galen
- learned about structure of relatively
advanced fetuses
- minute dimensions resisted serious
analysis
- development of microscope allowed
study of early stages

Special Fields in Embryology

de Graaf
- described ovarian follicles

Comparative Embryology
- Provided insight for concept that
ontogeny recapitulates phylogeny
- Recognition of different modes of
development
- Adoption of model organisms

Hamm and Leeuwenhoek


- first saw human sperm
Spermists vs Ovists
- spermists: sperm contained the new
individual in miniature and was
nourished in ovum
- ovists: ovum contained a minute body,
stimulated to grow by seminal fluid
o Bonnet:
discovered
parthogenetically
developed
insect eggs (supported ovists)
Spallanzani and Wolf
- Laid to rest spermist/ovist view
(preformation)
- Spallanzani: demonstrated that both
male and female sex products are
necessary
for
initiation
of
development
- Wolf:
Epigenesis,
embryonic
development
occurs
through
progressive remodeling and growth
von Baer
- von Baers law: the more general basic
features of any animal group appear
earlier in development than do the
special features that are peculiar to
different members of the group
- Germ layer theory: demonstrated
existence of germ layers in embryos
o Significance
could
not
be
grasped until cellular basis of
animal structure was known
Schleiden and Schwann
- Formulation of cell theory
Embryology
- Zygote has dual origin from two
gametes: spermatozoa and ovum
- Fertilization is starting point of life
history of individual (ontogeny)
- Period starting with fertilization until
metamorphosis, hatching, or birth
Weismann
- Germ cell theory
- Made distinction between soma and
germ-cell line

Descriptive Embryology
- Basic structural
embryonic body
- Serial
sections,
reconstructions

pattern
3D

wax

of

the
plate

Experimental Embryology
- Understand
causative
factors
in
development by posing hypotheses
and testing them by manipulation of
embryos
- Roux
o Experiment of 2 cell embryo
Destroyed
one
blastomere
Each cell is capable of
giving rise to complete
individual
Provided
proof
of
untenability
of
the
preformationist doctrine
Chemical Embryology
- Provided description about chemical
and physiological events in embryo
- Interaction between components and
how basic body pattern is laid down
Teratology
- Concerned
with
study
of
malformations
- Identify and eliminate genetic and
environmental factors that cause
congenital defects
Reproductive Biology
- Related to problems of conception and
contraception
- Emphasis
on
gametogenesis,
endocrinology, transport of gametes,
fertilization, embryonic development
Developmental Biology
- Embryonic development + postnatal
development and processes
- Focus on processes and concepts,
rather than specific morphological
structures
- Plant and animal systems included
Embryology in Contemporary Society
Test tube baby

In vitro
transfer

fertilization

and

embryo

The Cell and its Environment


Intracellular Synthesis and its Regulation
- Regulatory mechanisms restrict or
permit synthesis of specific proteins
and other macromolecules
- DNA (transcription) mRNA with
introns
and
exons
(mRNA
processing) definitive mRNA
(1,
formation
of
structural
proteins/enzymes) ribosome linkage
(2, secretory proteins) mRNA-rER
complex

Golgi
complex

membrane vesicles
- May
be
mediated
by
receptor
molecules located at cell surface
activated by binding of a ligand,
causing
stimulation
of
signal
transduction.
Cell Surface
- Junctional complexes
o Desmosomes
bind epithelial cells
focal
points
for
attachment of fibrillar
intracellular proteins
o Gap junctions
Mediates communication
and exchange of small
molecules
o Tight junctions
On surface of many
epithelia
Bind
adjacent
cells
together,
forming
impermeable barrier
Prevent
mingling
of
membrane proteins on
either side of junction
- Cell adhesion
o Experiment: disaggregated and
reaggregation of sponges
o Ca++ mediated adhesion
Glycoproteins
E-cadherin
(epithelium)
N-cadherin
(nerves,
mesoderm)
P-cadherin
(placenta)
o Heterophilic binding between
complementary saccharides
Occurs
during
mammalian fertilization,
when
head
of
spermatozoa encounters
zona pellucida
Extracellular Matrix
- Cells embedded on extracellular
matrix
- Collagen (glycoprotein with glycine)
o Basic unit is tropocollagen
o (I, II, III, IV, V, X)

Attachment glycoproteins involved in


attaching cells to other components of
extracellular matrix
o Fibronectin (I, III, V)
o Chondronectin (II, X)
o Laminin (IV)
Glycosaminoglycans
o Example: hyaluronic acid in
raising fertilization membrane
of the egg

Fundamental Processes and Concepts in


Development

than one type of tissue, depending on


inductive stimulus

Cell Division and the Cell Cycle


- Postmitotic cells further division does
not occur
- Maturation-promoting factor (MPF)
o Cdc2
Present throughout cell
cycle
Activation by cyclin and
dephosphorylation
Activation of MPF exerts
mitotic effects on cells
o Cyclin
Produced during G1
Broken down after mitosis

Differentiation
- Restriction and determination signify
progressive
limitation
of
developmental capacities
- Differentiation: actual morphological
or functional expression of a particular
cell or group of cells; the process
where cell is specialized
Morphogenesis
- Processes that mold external and
internal configuration of embryo
- Pattern formation: laying down of
morphological blueprint
- Morphogenesis: actual realization of
plans
- Homeotic
genes:
determine
the
regional
characteristics
of
each
segment (14 segments)

Gene Activation
- Derepression of heterochromatin or
repression of euchromatin
- Derepression of general genes from
zygote to blastula
- Derepression of tissue specific genes
from gastrula to organogenesis
Restriction and Determination
- Restriction:
the
reduction
of
developmental options permitted to a
cell
- At
gastrulation,
one
stage
of
restriction has occurred (endo, meso,
ectoderm)
- Part of ectoderm thickens and
undergoes Neurulation
- Determination
o restriction has proceeded to a
point where a group of cells
becomes committed to single
developmental fate
o final
step
in
process
of
restriction
- Inductions
(tissue
interactions)
precede
determination
(and
restriction)
Induction
- Form of embryonic signal calling
- Effect of one embryonic tissue on
another, so that the developmental
course of responding tissue is changed
from what it would have been in
absence of inductor
- First major inductive event: induction
of mesoderm in cleavage
- Primary
induction:
induction
of
nervous system during and shortly
after gastrulation
- Secondary induction: nervous system
induces other structures
- Permissive
induction:
inductive
signal
required
to bring about
development of structure
- Instructive induction: responding
tissue has options of forming more

Intercellular Communication
- Intercellular
communication
place in localized gap junctions

takes

Cell Movements
- Individual cells commonly migrate by
means of amoeboid movements
- Unique: in avian embryos, primordial
germ cells move from wall of yolk sac
into the bloodstream and are carried
via blood to the gonads
- Amoeboid movement examples:
o Ectoderm: migration of cells
away from neural crest
o Mesoderm: spreading out of
mesodermal cells
o Endoderm: migration of primary
germ cells from yolk sac to
gonads in mammals
- Movement as sheet seen in epithelial
cells
Cell Death (Apoptosis)
- Examples:
o Tail and opercular resorption
o Separation of digits
- May be hormonally controlled
o Male and female genital ducts
The Clonal Mode of Development
- Clones: group of cells arising from
single precursor
Regulation and Regeneration
- Regulation
o restoration of missing material
occurring before differentiation
of structure
o basis
for
development
of
identical twins
o subdivision of inner cell mass
- Regeneration
o Differentiation
of
structures
already occurred

Growth
- Differential growth: all parts of the
embryo do not grow at the same rate
- Determinate growth: body grows to
certain point that is characteristic of
species and sex (mammals)
- Indeterminate
growth:
ancestral
vertebrates (fish)
Recapitulation
- Biogenetic law of Muller and Haeckel
o Ontogeny is an abbreviated
recapitulation of phylogeny

REPRODUCTIVE
SEXUAL CYCLE

ORGANS

AND

THE

Reproductive Organs
Female Reproductive Organs
- Paired gonads (ovaries) located in
pelvic cavity
- Each ovary lies close to funnel-like
opening (ostium tubae) at end of a
uterine (fallopian) tube
- Around abdominal orifice of tube is
fimbriae
- Uterus:
thick,
vascular,
smooth
muscle, caudally continuous with the
cervix, will project into vagina
- Vagina: organ of copulation and birth
canal
Male Reproductive Organs
- Testes suspended in scrotum (with a
countercurrent
heat-exchange
system), with lower temperature
- Spermatozoa produced in seminiferous
tubules, then to tubuli recti, rete
testis, ductuli efferentes, epididymis,
ductus deferens
- Spermatozoa stored in epididymis and
ductus deferens
Sexual Cycle in Mammals
Estrous Cycle in Mammals
- Sexual cycle
o Estrus
(prepared
for
reproduction accompanied by
ovulation and sexual desire)
o Postestrum
(regression
of
preparations)
o Diestrum (period of rest)
o Proestrum (period of active
preparatory changes)
- Light is a critical factor: higher than
certain threshold causes hypophysis to
become active and produce enough
FSH
- Light Nervous transmission FSH
Follicular growth Estrogen in
ovarian follicle estrogen in blood
stream mating behavior, estrous
uterus, estrous vagina
Primate Menstrual Cycle
- Sexual cycle is characterized by
menstruation
- Commences
at
menarche
until
menopause
- Three phases
o Menses
o Proliferative (follicular)
o Secretory (luteal)
- Menstruation initiated by reduction of
blood flow into superficial uterine
blood
vessels,
resulting
in
deterioration and extravasation of
blood into tissue, bringing with it the
necrotic superficial tissue

Hormonal Regulation of the Female Sexual


Cycle
- Levels of hormonal control
- 1: Hypothalamus
o GnRH (stimulates LH and FSH in
hypophysis)
o PIH (inhibits prolactin release by
hypophysis)
- 2: Hypophysis
o FSH (stimulate follicle cells to
produce estrogen)
o LH (male: stimulate Leydig to
produce testosterone; female:
stimulate follicle to produce
progesterone)
o Prolactin
(promotion
of
lactation)
- 3: Ovaries (hormones secreted into
blood and placental tissues)
o Estradiol
o Progesterone
o Testosterone:
precursor
of
estrogen biosynthesis; induces
atresia
o Inhibin: inhibits FSH secretion
- 4: Ovarian steroid hormones into body
tissues
- Ovarian follicle maturation brought by
rise in pituitary FSH FSH and LH
stimulate follicle estrogen production
high estradiol production in ovarian
follicle causes LH and FSH peak LH
peak as final stimulus for follicular
maturation ovulation follicle
transforms into corpus lutem (actions
of LH) corpus luteum secretes
estradiol
and
progesterone

increased ovarian hormone levels and


inhibin cause feedback inhibition
inhibition results in low levels of LH
and FSH later regression of follicle
comes decrease in gonadotropin levels

stimulation
of
GnRH
and
gonadotropins
Hormonal Regulation of Reproduction in the
Male
- Testosterone: secreted by Leydig cells
(stimulated by LH)
- Sertoli cells take up FSH synthesis
of ABP

GAMETOGENESIS
Gametogenesis
- Germ plasm gametes + cells that
give rise to them
- Gametogenesis:
germ
plasm
is
converted to specialized sex cells
capable of uniting at fertilization and
producing a new being
- Four phases
o Origin of germ cells and
migration to gonads
o Multiplication of germ cells in
the gonads (mitosis)
o Reduction
of
number
of
chromosomes (meiosis)
o Maturation and differentiation of
gametes
The Origin of Primordial Germ Cells and Their
Migration to the Gonads
- Primordial germ cells of mammals,
reptiles, birds arise in epiblast take
up
temporary
residence
in
extraembryonic
tissue
before
returning
o Birds: in germinal crescent
o Mammals: posterior wall of yolk
sac (near allantois)
o Amphibians:
vegetal
pole
cytoplasm
- Note: PGCs do not produce gonads,
they produce gametes
- PGCs in extragonadal sites may
develop into teratomas
Proliferation of Germ Cells by Mitosis
- Mitotically active germ cells: oogonia,
spermatogonia
- Settling in the gonads induce a
proliferative phase
- Mammalian
oogonia
proliferative
phase
o Mitosis brings numbers to about
7 million at 5th month of
pregnancy
o Atresia causes sharp decline
o Primary
oocytes
formed
(suspended at prophase I)
Meiosis
- Genetic recombination occurs by
o Random
distribution
of
chromosomes to daughter cells
o Crossing over
- (2n, 4c) (Me I) (1n, 2c) (Me II)
(1n, 1c)
Spermatogenesis and Oogenesis Compared
- 4 functional spermatozoa vs 1 viable
ovum
- Arrests in meiosis
o Spermatogenesis: none
- Oogonesis meiotic arrest
o First
arrest
at
diplotene
prophase I in primary oocyte
o Broken by hormonal changes

o
o

Arrested again at metaphase II


Broken by fertilization (sea
urchins complete meiosis II at
once, no second arrest)

Spermatogenesis
- Transition from mitotically active PGCs
to mature spermatozoa
- Three phases
o Mitotic multiplication
(spermatocytogenesis)
o Meiosis (spermatidogenesis)
o Maturation and differentiation
(spermiogenesis)
- Spermatogonia
o Type A: stem cell population
Ad: long term reserves
Ap: mitotically active,
give rise to B
o Type B: committed to finish
spermatogenesis
- Spermatocytes
o Preleptotene spermatocytes
o Primary spermatocytes (Me I)
secondary spermatocytes (Me
II) haploid spermatids
- Differentiation
of
spermatid
to
spermatozoa
o Golgi
complex
forms
proacrosomal
granules, into
acrosome
o Centrioles as point of anchorage
for developing flagellum
- Intercellular
bridges:
facilitate
synchronous
differentiation
and
division of sperm-producing cells
- Sertoli cells:
o FSH target Sertoli cells
o Synthesis of ABP (to maintain
high testosterone levels)
o Maintain blood-testis barrier
o Create
environment
for
differentiation of sperm cells
o Facilitate release of mature
spermatozoa
o Degradation of residual bodies
- Blood-testis barrier (held by tight
junctions) responsible for preventing
bodys
immune
system
from
destroying
mature
sperm
cells
(antigenically different from body)
Gene Expression during Spermatogenesis
- Posttranscriptional control
Sperm Maturation
- Sperm coated with glycoprotein which
must
be
removed
in
female
reproductive tract before fertilization
can occur (activation, capacitation)
- Seminal fluid provides external energy
source (causes nonmotile sperm to
become motile)
Oogenesis
Oogenesis in Amphibians
- Mitotic phase of oogenesis does not
come to early halt
- New crop of eggs each year, 3 years
for maturation

Follicular epithelium, theca, ovarian


epithelium

Development of the Amphibian Egg


- Three phases
o Previtellogenesis (before yolk
deposition)
o Vitellogenesis (period of yolk
deposition)
o Maturation
(released
from
meiotic block by progesterone)
- Previtellogenic phase
o Period upto early diplotene
phase of meiosis
o Lampbrush
chromosomes:
spread
out
configuration
forming loops
Loops are where RNA
synthesis occurs
o Large numbers of nucleoli for
specific
gene
amplification
(ribosomes, rRNAs)
- Vitellogenic phase
o Principally concerned with yolk
formation
o Lipochondria stores lipids
o Glycogen granules stores
carbs
o Yolk platelets stores proteins
Yolk protein produced by
liver cells under estrogen
influence
Gonadotropin
release
from hypothalamus to
oocyte

estrogen
secretion from oocyte
into liver secretion of
vitellogenin from liver to
oocyte
o Yolk precursor vitellogenin
(phospholipoprotein)
Incorporated into oocyte
by micropinocytosis
Represented
by
phosvitin
and
lipovitellin (packed in
crystalline form to form
yolk platelets)
o Yolk formation was thought to
be function of Balbiani body
(yolk nucleus)
o Pigment granules concentrate
at animal hemisphere
- Maturation phase
o Hormonally induced release of
egg from first meiotic block
o Breakdown of germinal vesicle
o Completion of first meiotic
division
o Formation of first polar body
o Begins
with
secretion
of
progesterone (stimulated by
gonadotropin)

Causes breakdown
of
germinal vesicle, meiotic
maturation
Meiosis
arrested
again
at
Metaphase II by CSF (cytostatic
factor)

Oogenesis in Birds
- Yolk is a single cell (the ovum)
o Gradually
accumulates
in
cytoplasm of ovum before it is
liberated from ovary
o All other noncellular secretions
(egg white, shell membrane,
shell) are contributed as ovum
passes reproductive tract
- Yolk still produced by the liver and
transported via blood to the follicular
cells
surrounding
ovum
(as
in
amphibians)
o 50% water, 33% fat, 16%
protein, 1% carb
o Water NaCl, Ca salts (bone
formation)
o Proteins lipovitellin (binds w/
lipids), phosvitin (binds w/
phosphorus)
- Protuberance containing ovum is
ovarian follicle
- With zona radiata (irregular striated
plasma membrane due to microvilli),
for increase in membrane surface,
enhancing metabolic interchanges
- Compared with mammalian ovum:
o No yolk in mammals, just liquor
folliculi
o Both have two layered CT theca
- Yolk release albumen secretion
shell membranes (isthmus) shell
(uterus)
Oogenesis in Mammals
- Primary oocyte (so called as it is
undergoing meiosis I) + flat follicular
cells = primordial follicle
- Phase I: pool of primordial follicles
developing into primary follicles (flat
cuboidal)
- Meiotic
arrest
follows
(diplotene
meiosis I)
- Both oocytes and follicular (granulosa
cells) develop microvilli, connected by
gap
junctions
(allow
high
MW
molecules to pass through)
- Zona pellucida beings to develop
- Phase II: growth of oocyte and
granulosa covering (under influence of
gonadotropic hormones)
- Overall growth of follicular covering
mediated by FSH receptors
- Secondary follicle when antrum is
identifiable
- LH
receptors
develop,
allowing
production of testosterone by theca
o Transported into granulosa cells,
wherein
granulosa
converts

testosterone to estrogen by
aromatase
Phase III: further follicular growth and
selection of one follicle (highest
receptivity to FSH) which will undergo
ovulation
o Begins late in follicular phase of
menstrual cycle
o After LH surge, angiogenesis
occurs, causing estradiol to spill
out into blood
o Before
ovulation,
ovum
is
released from first meiotic block
(diplotene), allowing meiosis I to
finish
o After that, follicle is now ready
to respond to preovulatory FSH
and LH surge, and release itself
(wherein it is now at metaphase
II, second block)
Ovulation
o Increased antral fluid pressure
within follicle causes bursting of
follicular wall
o Weakening of follicular wall by
lytic enzyme (stimulated by LH)
Corpus luteum
o Stratum granulosum and theca
interna involved in corpus
luteum formation
o Endocrine
organ,
secreting
progesterone and estrogen
o Granulosa
cells
swell
and
develop to secrete high levels of
hormones
o Formation of corpus lutem
require continuous presence of
LH from pituitary
regression happens with
decreased sensitivity to
LH receptors
corpus
lutem
of
pregnancy maintained by
chorionic
gonadotropin
(secreted by embryo)
o Corpus lutem produces large
amounts of progesterone and
estrogen
Progesterone
prepares
lining
of
uterus
for
implantation

Accessory Coverings of Eggs


Covering of the Sea Urchin Egg
- Inner to outer
o Plasma membrane
o Vitelline envelope (composed of
glycoproteins, contain speciesspecific
receptors
for
spermatozoa)
o Jelly
coat
(polysaccharides,
glycoproteins,
hydrates
and
expands when eggs are shed)

The Membranes Surrounding the Amphibian


Egg
- Plasma membrane: forms microvilli
- Follicular cells: form macrovilli
- Narrow space between oocyte and
follicular epithelium becomes filled
with noncellular basement membrane:
vitelline envelope (equivalent of zona
pellucida in mammals)
- Gap
junctions
join
the
villous
processes
- At ovulation, perivitelline space forms
between
vitelline
envelope
and
plasma membrane
- Coated with jelly coat as it goes
through oviduct (same function as sea
urchins)
Formation of the Accessory Coverings of Bird
Eggs
- At ovulation, ovum surrounded by
inner vitelline membrane
- Remainder of accessory coverings
secreted about ovum during passage
toward cloaca
- Outer vitelline membrane laid down
when it is in oviduct adjacent to ovary
- Albumen laid down in upper oviduct
o Rotation twists albumen into
spiral strands at two ends of
yolk: chalazae
Serve to suspend yolk in
albumen
- Egg white
o Ovalbumin and lysozyme by
estrogen
o Avidin secreted by goblet
cells by progesterone
- Shell membrane added farther along
oviduct
- Shell secreted as egg passes through
shell gland (at uterus)
The Coverings of Mammalian Eggs
- Noncellular zona pellucida (mostly
synthesized by oocyte)
o ZP-1
o ZP-2
o ZP-3 acts as sperm receptor
and plays a role in inducing
acrosome reaction
- Corona
radiate
still
surrounds
mammalian ovum (may continue to
secrete steroids and prostaglandins)

The Sea Urchin


Gamete Release and Transport
- 100 billion spermatozoa and 4 billion
eggs

Sperm
Penetration
of
the
Egg
in
Invertebrates and the Acrosome Reaction
- When spermatozoa encounter egg,
former undergoes changes
- In presence of egg cells, spermatozoa
will cluster and increase motility
- Direct contact with jelly coat increases
motility and stimulates acrosome
reaction
- Speract is responsible for increased
motility and activated respiration that
occur when sperm contacts with jelly
coat
o Increase in permeability of
plasma causes influx of Na and
Ca, efflux of H+
Raises intracellular pH,
stimulating
flagellar
activity
- Contact w/ jelly coat stimulates
acrosome reaction
o Begins with breakdown and
subsequent fusion of outer
acrosomal
membrane
and
plasma membrane
o Polymerization of G-actin to Factin
(forming
acrosomal
process)
Tip of process is covered
with bindin, mediating
sperm binding to surface
of eggs
o Spermatozoa digest through
vitelline membrane by lysins
Binding of Sperm to the Egg
- Sperm receptor molecule (on microvilli
of egg)
o Intracellular: remains constant
among species
o Extracellular: differs accdg to
species
- After sperm-egg fusion, fertilization
cone forms by microvilli engulfing
sperm head
Blocks to Polyspermy
- Fertilization of egg by more than one
sperm
- Fast block

Membrane event, set in place


within 2-3 seconds, and lasts for
60 seconds
o Acrosomal process and plasma
membrane
fusion
causes
depolarization
of
plasma
membrane (by influx of Na+)
From -70mV to +10mV
Positive potential does
not permit fusion of other
spermatozoa to plasma
membrane
Slow block
o Mobilization of Ca from within
egg
o first released at site of sperm
entry
o wave of released Ca initiates
cortical reaction (rupture of
cortical granules and release of
these contents into perivitelline
space)
o cortical granules move to inner
surface of plasma membrane,
fuse with it, and open up
o contain
sulfated
mucopolysaccharides (GAGs),
which have high water affinity,
causing swelling and forcing
vitelline envelope away from
plasma membrane (raising the
fertilization membrane)
o Fertilization
membrane:
name
given
to
vitelline
envelope after changes by
cortical reaction
o Hydrated mucopolysaccharides
form hyaline layer (between
plasma
and
fertilization
membrane)
o As fertilization membrane is
elevated, an enzyme alters it,
causing attached sperm to drop
off
o Final
step:
release
of
ovoperoxidase from cortical
granules (for breakdown of
H2O2, results in hardening of
fertilization membrane)
H2O2 released by egg
during cortical reaction
(spermicidal)
Polyspermy is normal in urodele
amphibians and birds
o

FERTILIZATION
- Initial contact between egg and sperm
- Entry of sperm cell into egg
- Prevention of polyspermy by egg
- Metabolic activation of egg
- Completion of meiosis by egg
- Formation and fusion of male and
female pronuclei

Metabolic Activation of the Egg


- Other events that prepare egg for
fusion of genetic material:
o Increased
oxygen
consumption
o Activation of NAD kinase
(facilitate biosynthesis of new
membrane lipids)
o Another influx of Na+ (with
efflux of H) causing increased
intracellular pH

Increased pH leads to increased


protein synthesis and initiation
of DNA synthesis

Penetration of the Spermatozoon into the


Egg and Fusion of the Genetic Material
- Sperm nucleus begins to interact with
egg cytoplasm, and chromatin relaxes
- As
chromatin
dispersion
nears
completion, new membrane forms
around what can now be called a male
pronucleus
- Sperm centrioles persist, and provide
basis for formation of sperm aster,
important in getting male and female
pronuclei together
- Pronuclear fusion occurs at center
of egg
- After fusion, chromosomes replicate in
preparation for cleavage

Mammalian Fertilization
Sperm Transport in the Female Reproductive
Tract of Mammals
- Barriers to fertilization:
o Natural
acidity
of
vagina
(bacteriostatic)
Seminal fluid acts as
buffer (raises vaginal pH)
Seminal fluid may cause
contractions
in
upper
vagina, helping propel
spermatozoa
Orgasms cause uterine
contractions
o Entrance
to
uterine
tubes
(ovulation can sometimes only
occur on one tube)
- Positive rheotactic response (face
an oncoming current generated by
uterine ciliary movement)
- Capacitation:
removal
of
glycoprotein covering spermatozoa,
enabling better penetration of egg
Egg Transport
- Ciliary currents and smooth muscle
contractions transport egg into uterine
tube
- Corona radiata adds mass faster
movement down tube
Union of Gametes
- Mammals: occurs in upper part of
uterine tubes
- Spermatozoa must penetrate corona
radiata cells and then zona pellucida
before contact with plasma membrane
- Zona pellucida: molecules on sperm
head bind with species-specific sperm
receptors (consist of exposed part of
ZP-3)
o Further contact with other core
regions
stimulate
acrosome
reaction
(capacitation
is
prerequisite,
so
that
lytic
enzymes
are
released
to
facilitate passage of sperm
through zona pellucida)
- Acrosome reaction:
o Localized fusion and breakdown
of outer acrosomal membrane
and plasma membrane
o Acrosin
bound
to
inner
acrosomal membrane digests
through zona pellucida
o Fertilization cone bulged out
when sperm makes contact with
egg
Development and Fusion of Pronuclei
- With sperm entry, block to second
meiotic division is lifted, and second
polar body is released
- Breakdown
of
sperm
nuclear
membrane and decondensation of

chromatin and protamine replacement


with histones
- New pronuclear membrane forms
around decondensed material
- DNA synthesis occurs as male and
female pronuclei migrate toward each
other (as opposed to sea urchins,
where chromosomes condense to
prepare for metaphase, and DNA
synthesis occurs after fusion of
pronuclei)
Parthenogenesis
- Activation of unfertilized eggs and
development into viable individuals
- Mammals: all female because females
are homogametic (XX)
- Birds, reptiles: males and females
(heterogametic females)
Sex Determination
- Occurs at fertilization, determined by
o Mammal sperm
o Bird/reptile egg
Establishment of Polarity in the Embryo
- 3 polar axes:
o Craniocaudal (anteroposterior)
o Dorsoventral
o Mediolateral
Establishment of Polarity in Amphibians
- Primary polarity of egg by animal
and vegetal poles
o Denser pigment concentration
at animal pole
o Nucleus located near animal
pole
o Gradient of increasing density
of ribosomes and glycogen
granules towards animal pole
o Size and concentration of yolk
platelets
increase
toward
vegetal pole
o Region of animal pole = head
o Region of vegetal pole = tail
Anterocaudal axis
- Fertilization is next establishment of
polar axes
o 2 major reorganizations
o General
convergence
of
cytoplasm beneath the thin
cortical region toward the
sperm entry point
o 30
degree
shift
between
subcortical
cytoplasm
and
overlying cortex
o These changes cause reduction
in density of pigment granules
in the region of the animal
hemisphere along equatorial
zone opposite to sperm entry
point
Reduced
pigmentation
called gray crescent
Midpoint of gray crescent
is middorsal point of

body
(determines
dorsoventral axis of
future embryo)
Determination of two axes determines
third
Even before cleavage, the three axes
are
established
and
secondary
polarization is completed

Establishment of Polarity in Birds


- Cleaving embryo represented by flat
disk of cells on yolk surface: cells on
outer surface become dorsal part,
those closest to yolk become ventral
- Cells are shed from part of blastoderm
uppermost (with respect to gravity).
The area from which these cells fall
become the caudal end of embryo

CLEAVAGE AND FORMATION OF THE


BLASTULA
- Cleavage: waves of cell division
following one another almost without
pause
- Many of changes and differences in
cleavage patterns among embryos of
various species are related to the
amount of yolk present in egg
- Cleaving embryo develops a central
cavity (blastocoel) and enters blastula
stage
The Cell during Cleavage
- Cleavage
division
consists
of
karyokinesis followed by cytokinesis
- Cleavage furrow first forms in the
region of cortex nearest to mitotic
spindle, and then moves around cell
- Position of cleavage furrow gets fixed
or established at anaphase
- Asters (composed of microtubules)
interact with cell cortex to stimulate
cleavage furrow formation
o Asters are the effective agent
in initiating cleavage furrow
formation (not the mitotic
spindle)
o Mitotic spindle asters cell
cortex
Distribution of Yolk and its Effect on
Cleavage
- Blastomeres: cells that arise from
cleavage
- Holoblastic cleavage: characterized by
complete division of cells
o Oligolecithal
eggs
produce
blastomeres of equal size
o Mesolecithal
eggs
displace
nucleus towards animal pole
Net result is appearance
of
later
and
larger
blastomeres at vegetal
pole
- Meroblastic cleavage: newly formed
plasma membrane does not separate
inner borders of dividing cells from the
underlying yolk
o Telolecithal eggs: displaces the
embryo-forming cytoplasm into
a small disk on one edge of
ovum
Cleavage and Formation of the Blastula
Amphioxus
- Radial holoblastic equal cleavage,
microlecithal eggs
- Meridional meridional equatorial
meridional equatorial
Sea Urchins
- Early cleavage only consists of S and
M phase, with alternating periods of
cyclin synthesis and degradation

G1 and G2 phases evident only later in


cleavage
Microlecithal
Meridional meridional equatorial
o Lower tier undergoes unequal
equatorial division
Micromeres
(becomes
primary mesenchyme)
Macromeres
o Upper tier undergoes equal
meridional division, forming 8
mesomeres
During entire cleavage, embryo is
enclosed in fertilization membrane
and closely associated with hyaline
layer
Blastomeres later form motile cilia,
penetrating
hyaline
layer
and
secreting hatching enzyme into
perivitelline space, digesting the
fertilization membrane
End
product
is
mesenchyme
blastula

Amphibians
- Meridional (begins at animal pole,
bisects gray crescent) meridional
equatorial double meridional
double equatorial
- Amphibian blastocoel
o Formed from specialization of
cleavage furrow of animal
hemisphere
o Filled with Na+ ions, then water
enters
to
maintain
ionic
balance, causing expansion
- Amphibian blastula has three main
regions
o Region around animal pole,
including cells forming roof of
blastocoel
Future ectodermal germ
layer
o Region around vegetal pole,
including large cells in interior
Future endodermal cells
o Marginal ring of cells in the
subequatorial region of the
embryo, including gray crescent
Embryonic mesoderm
- Nieuwkoop
discovered
basis
for
mesodermal induction
o Experiment
1:
directly
apposed sheet of cells from
animal
hemisphere
above
blastocoel to a yolk mass from
vegetal hemisphere
Inductive
influence
from yolk mass caused
animal pole cells to form
mesodermal
structures
Therefore,
blastocoel
may
function to restrict
interaction

between
future
endodermal
and
ectodermal cells
o Experiment 2: isolated pieces
of ectoderm induced to form
mesoderm by transforming
growth factor-beta
Source of mesodermal induction
(Nieuwkoop center or dorsalizing
center
or
mesodermal
inducing
center), resides in a number of vegetal
endodermal
cells
located
in
prospective dorsal midline
Nieuwkoop center
o Stimulates
formation
of
mesoderm
o Establishes dorsal properties of
induced mesoderm
Dorsal induced mesoderm is direct
forerunner of what is called the
Spemann organizer (dorsal lip of
blastopore), the dominant organizing
region of amphibian embryo during
gastrulation

Birds
- Mitotic spindles align themselves so
subsequent cleavage furrows form at
right angles to the preceding one (first
3)
- Fourth
cleavage
furrow
is
a
circumferential one
- Blastomeres formed by first few
divisions are dorsally bound by plasma
membrane but basal surfaces are
open to underlying yolk
- Further cleavage of blastoderm
results in radial extension of embryo
- 32-cell
embryo
shows
cleavage
parallel
to
surface,
establishing
several strata of superficial cells
- At around 100 cells, blastoderm is
underlain by a subgerminal cavity
o pH of subgerminal cavity 6.5,
while albumen: 9.5, leading to
establishment of transepithelial
potential of 25mV. Electrical
gradient
determines
dorsoventral
axis
of
blastoderm.
- Shedding of cells begins from
undersurface of area of blastoderm
that is farthest away from source of
gravity (this area becomes caudal
end of embryo)
o Area pellucida: central portion
of blastoderm thinned out by
shedding of cells
o Area opaca: region where
blastoderm cells still abut
directly onto yolk
- Primary hypoblast: aggregates of
cells shed from lower surface of
blastoderm
by
a
process
of
polyingression/delamination

First occurs at posterior end of


embryo
o Separated from epiblast by a
blastocoel
o Polarity and location of primary
hypoblast determine location
and
direction
of
future
primitive streak by inductive
interaction
o EXTRAembryonic endoderm
Comparison with amphibians
o Two layered blastoderm is
compared
to
a
flattened
blastula
(epiblast:
animal
hemisphere::
hypoblast:
vegetal)
o Both hypoblast and vegetal pole
have ability to induce formation
of mesoderm in epiblast and
animal
pole
by
inductive
interaction
o

Mammals
- Equal holoblastic cleavage of an
isolecithal egg
- Persistence of traits characteristic to
large-yolked
embryos
(later
in
development)
- Second cleavage division may not
occur
simultaneously
in
both
blastomeres
o Mitotic
spindle
of
one
blastomere may rotate 90
degrees, causing a rotational
pattern of cleavage
(as
opposed
to
radial
in
echinoderms, Amphioxus)
- 16-cell stage still is contained in zona
pellucida
- Morula: internal secretion of fluid by
blastomeres leads to formation of
blastocoel, or blastocyst cavity
o Similar to amphibian (Na-KATPase system brings in Na, and
then water)

Blastocyst has two populations of cells


o Trophoblast:
cells
that
constitute
outer
wall
of
blastocyst
maternal X chromosomes
are
preferentially
expressed
assumed
epithelial
properties
(tight
junctions, microfilaments)
forms
large
part
of
placenta
o Inner
cell
mass:
joined
together by gap junctions,
retains ability to reaggregate if
separated
Comparison with birds
o Embryos of mammals form a
layer of cells beneath inner cell
mass,
called
primitive
endoderm
equivalent
to
primary hypoblast of avian
embryos
Note
that
primitive
endoderm
does
not
contribute
to
embryo
proper,
as
primary
hypoblast does not too

GASTRULATION AND THE FORMATION


OF THE GERM LAYERS
Gastrulation as a Process
- Well-ordered rearrangements of cells
in embryo
- Morphogenetic movements result in
reorganization
- Rearrangement of blastula to a stage
characterized by presence of three
germ layers
Table 6-1
Type
movement
Invagination

of

Evagination
Involution

Epiboly

Ingression

Polyingression
(delamination)

Ameboid motion

Description
Inpocketing
sheet of cells

Example
of

Outpocketing of
sheet of cells
Rolling around a
corner
of
an
expanding outer
layer of cells and
spreading over an
internal surface
Spreading of a
cell sheet

Sinking
of
individual
cells
from a surface
into an area
Separation
of
second
sheet
from an original
single sheet
Migration of cells
as
single
individuals
through their own
motility

Archenteron
formation
in
Amphioxus
Exogastrulation
Cell movements
through
the
amphibian
blastopore

Spreading
of
outer
cells
towards
amphibian
blastopore
Primary
mesenchyme
formation in sea
urchin embryos
Formation
of
primary
hypoblast
of
avian embryos
Migration
of
neural crest cells

Two main strategies for gastrulation:


o Carry
out
gastrulation
movements within context of a
sphere
(Amphioxus,
amphibians)
o Elaboration of three germ layers
as two-dimensional sheets upon
one sector of an enormous
sphere (birds, reptiles, even
mammals)
Blastopore: opening from outside
into the archenteron

Gastrulation in Sea Urchin Embryos


- Separation of primary mesenchyme
from vegetal plate of blastula signifies
start of gastrulation
- Primary mesenchymal cells develop
filopodia, moving along the basal
lamina until forming a ring-like
structure near base of invaginating
archenteron
- Three
stages
of
formation
of
archenteron
o Invagination of cells at vegetal
pole
o Presence
of
secondary
mesenchyme (will later make
contact with animal pole by
filopodia)

As tip of archenteron makes


contact
with
animal
pole,
secondary
mesenchyme
undergoes final determination
(can no longer dedifferentiate
into primary mesenchyme cells)
Pluteus larva
o

Gastrulation in Amphibian Embryos


- Cortical rotation, stimulated by sperm
fusion, leads to generation of an early
organizing
center
(Nieuwkoop
center) in dorsal cells of vegetal
hemisphere
o Again, major activity of vegetal
organizing
center
is
mesodermal induction
- As embryo enters late blastula,
organizing activity shifts to a more
superficial dorsal location
o Under genetic influence, cells of
dorsal marginal region migrate
resulting
in
formation
of
blastopore
o Upper margin of blastoporal
groove is known as dorsal lip
of blastopore and will later
become the major organizing
region (Spemann organizer)
- Initial formation of blastoporal groove
is related to a change in cell shape in
this area; cells elongate inwardly,
bottle cells, associated with inward
pulling movement, results in formation
of blastoporal groove.
- Prospective
endoderm:
around
ventral margins of blastopore and
extending to ventral part of embryo,
and lines archenteron
- Chordamesoderm:
passes
over
dorsal lip of blastopore and gives rise
to notochord and cephalic mesoderm
- Cell movement in dorsal lip of
blastopore:
o Surface cells are underlain by a
deeper marginal zone consisting
of several layers of cells
o Movement
associated
with
convergence and extension
phenomena
Account
for
overall
elongation of embryo
o Early gastrulation: cells of deep
layer
of
marginal
zone
interdigitate
by
radial
intercalation to form single
layer
o Late gastrulation: mediolateral
intercalation, where lateral
cells insert processes between
cells in medial part of layer and
later
become
interposed
between them
o Involuted cells from marginal
zone will form mesoderm of
embryo
- In surface layers, epiboly happens by
cell division, flattening, and spreading.
As these surface cells cross dorsal lip,
they become endodermal lining of
archenteron.
- Ectoderm:
neural
ectoderm
and
general cutaneous ectoderm

At end of gastrulation, all future


endoderm and mesoderm lie within
- Spemann and Mangold: demonstrated
that dorsal lip of blastopore possesses
ability to organize future development
o Early, it initiates convergent
extension movements
o Late, it exerts dorsalizing effect
on
lateroventral
region
of
marginal zone
o Organizer signals neighboring
cells of animal cap to form
neural plate
Gastrulation in Birds
- Review:
o Blastula
as
two-layered
structure
(epiblast
and
hypoblast, with blastocoel in
between)
o Embryo proper occupies area
pellucida and surrounded by
area opaca
- Kollers sickle (thin sickle-shaped
mass of cells) at posterior end of
embryo,
where
a
secondary
hypoblast pushes anteriorly, folding
primary hypoblast ahead of it
o Distribution of primordial germ
cells along anterior border of
blastoderm due to compression
of primary hypoblast (PGCs are
found in primary hypoblast)
- Gastrulation
begins
with
condensation of cells in posterior
part of epiblast, gradually assuming
cephalocaudal elongation, eventually
forming primitive streak
- Appearance of primitive streak is
result of inductive interaction of
epiblast with hypoblast
- Primitive streak now contains:
o Primitive groove: central furrow
o Primitive
ridges:
thickened
margins
o Hensens node: cephalic end
of streak
Begins to regress after
18hrs, with corresponding
elongation of notochord
above it (head process)
- Embryonic germ layers are formed
by migration of cells in epiblast
towards
primitive
streak,
and
subsequent
ingression
(not
delamination) to form middle and
lower
germ
layers
(embryonic
mesoderm and endoderm)
o First cells to pass are the future
embryonic endodermal cells
They displace primary
hypoblast outward and
cephalad toward area
opaca
o Mesodermal
cells
migrating
through:

Hensens
node:
notochord
Cranial part of primitive
streak:
embryonic
mesoderm
Caudalmost
part
of
streak: extraembryonic
mesoderm

Comparison of Avian and Amphibian


Development
- Blastula
o Bird:
epiblast
contains
endodermal and mesodermal
germ layers
o Amphibian:
surface
layers
contain
endoderm
and
mesodermal parts
- Induction
o Effect of hypoblast on epiblast
o Mesoderm induction by vegetal
yolk
- Blastopore
o Pre-primitive streak thickened
area of chick blastoderm
o Blastopore in amphibians
- Cell migration:
o Cells
that
constitute
endodermal layer migrate to
interior first, and mesodermal
layers follow later
Origin of the Germ Layers in Mammals
- Review:
o Mammalian
blastocyst
segregated into embryo forming
inner
cell
mass
and
trophoblastic cells
- Hypoblast: first cells to segregate out
from ICM
o Forms
extraembryonic
endoderm, similar to birds
o Forms lining of yolk sac
- Remainder of ICM is now called
epiblast
o Contains future ectodermal cells
o Also contains cells that will
migrate
through
primitive
streak and become definitive
endodermal and mesodermal
germ layers
- Remaining ICM now called embryonic
disk
o One margin of disk becomes
thickened, becomes caudal
part of embryo
From caudal thickening,
cephalad expansion of
cells results in primitive
streak
Origin of the Germ Layers in Rodents
- Early hypoblast is called primitive
endoderm in mouse
o Cells from this layer spread out
beneath
trophoblast

(trophectoderm)
to
form
endodermal layer of parietal
yolk sac
o Parietal endoderm cells create
basement membrane called
Reicherts membrane
o Polar
trophectoderm:
overlying ICM
can
undergo
mitosis,
daughter cells become
mural
o Mural
trophectoderm:
surrounding blastocyst cavity
Mitosis
results
in
polyploidy giant cells
ICM
undergoes
transformation
different from other mammals
o Protrudes deeply into blastocyst
cavity (like a tongue-like lobe)
o Cavity forms within the lobe
(proamnion),
and
cells
surrounding it are primitive
ectoderm (or epiblast)
Called an inverted egg
cylinder

NEURULATION AND THE FORMATION OF


AXIAL STRUCTURES
Primary (Neural) Induction

The Mesoderm of the Early Embryo


Secretion of Extracellular Materials in
the Early Embryo

Neurulation in Amphibians
Formation of the Neural Tube
The Neural Crest

The Formation and Differentiation of


Somitomeres and Somites