DEVBIOL
LECTURE 3: FERTILIZATION
Gliceria B. Ramos| 1st TERM | A.Y. 2022-2023
OUTLINE ○ Actual fusion of the plasma membranes of
I. A IV. B
the oocyte and sperm
II. ● Movement of sperm head into the cytoplasm of the
oocyte
prevents polyspermy; preventing other sperms from
● Causes cortical reaction binding to and entering the egg
○ Bursting of the cortical granules releasing
their chemical contents into the perivitelline
space
■ Space between the vitelline layer
and plasma membrane
○ Vitelline layer is lifted off from the plasma
FERTILIZATION membrane
● Beginning of a new organism ■ Forms the fertilization envelope
TYPES
● External (ex vivo)
○ Aquatic vertebrates (fish and amphibians)
○ Spawning is the release of eggs into the
aquatic environment
○ Challenge: ensure species-specific attraction
between sex cells
○ Solution: chemoattractant released by jelly
coat of oocyte (for sperm attraction and
activation)
● Internal (in vivo)
○ Avians and mammalians
○ Insemination is the deposition of sperm
cells into the female reproductive tract
○ Fertilization takes place in the oviduct or CORTICAL REACTION
uterine/fallopian tube ● Formation of the fertilization coat
● Fusion of the plasma membranes
MAJOR EVENTS IN FERTILIZATION ● Cortical reaction
● Contact and recognition between sperm and oocyte ○ Releasing of the contents (proteases, leuko
● Regulation of sperm entry into the oocyte polysaccharides, peroxidases)
○ Prevention to polyspermy ■ Proteases - break down molecular
■ Prevention of the entry of more than bonds between the vitelline
one sperm into the oocyte envelope and plasma membrane
● Fusion of the genetic materials of sperm and oocyte ■ Leuko polysaccharides -
○ Amphimixis or pronuclear fusion carbohydrate type, produce
● Activation of oocyte metabolism to start development osmotic gradient causing water to
○ Phases: Early Response of the oocyte and rush between the vitelline envelope
Late Response of the oocyte and the plasma membrane
■ Peroxidases - enzymes that
harden the fertilization membrane
FERTILIZATION PROCESS by cross linking tyrosine residues of
● Most knowledge was based on sea urchin fertilization adjacent proteins, hardening
● The Hertwig Brothers - Oscar and Richard prevents the attachment of other
○ Discovered fertilization using sea urchin as sperm to the oocyte
model organisms ● In summary, it is the breakdown of cortical granules
● Vitelline envelope leading to the release of chemical substances forming
○ Outer the cell membrane the fertilization membrane
● Jelly coat
○ Secrete the chemoattractants for
species-species recognition of sex cells in
the aquatic environment
1.CONTACT AND RECOGNITION BETWEEN SPERM
AND OOCYTE 2.REGULATION OF SPERM ENTRY INTO OOCYTE
● Noted structures: ● 1st block
○ Egg plasma membrane ○ Electrical in nature
○ Vitelline layer ○ Fast
■ Contains the species specific ○ Not sufficient enough
receptor for the sperm
○ Jelly coat ○ Plasma membrane fusion happens firstly
○ Sperm ○ The sperm acting under receptors on the
○ Sperm head plasma membrane of the oocyte, the oocyte
○ Acrosome is then activated
● Acrosome reaction ■ Caused by membrane
○ Starts with the breakdown of the acrosomal depolarization
cap, releasing the hydrolytic enzymes ■ Na+ channels open during
○ Hydrolytic enzymes degrade the jelly coat activation → Na+ influx into the cell
○ Jelly coat is digested ○ Cell at rest has a membrane potential of
○ Fibrillar protein (-50) to (-70) mV
■ Further penetrate the vitelline layer ■ More negative inside the cell than
● Fertilization cone outside
○ Activation (Na-Ca influx)
1
■ Fast reversal of the electrical
potential
■ becomes (-70) to (+10) inside the
cell – fertilization potential
■ Inside of cell is now more positive
than outside
● 2nd block (slow block to polyspermy)
○ Chemical in nature
○ Lasts longer
○ Backs up the 1st block to polyspermy
○ Starts w/ fusion
○ Activation of phospho-lipase C stimulation
and egg cell plasma membrane →
production of inositol trisphosphate and
diacylglycerol
■ Inositol trisphosphate
● Ca2+ is released →
cortical reaction
(breakdown and release of
4.METABOLIC ACTIVATION OF THE OOCYTE
substances) → formation
● Based on sea urchin studies
of fertilization cone
● fast block and cortical reaction are an early response
■ Diacylglycerol
● fusion → phospholipase C activation → Ca2+ release
● Early metabolic activation of the oocyte
→ NAD+ kinase → phosphorylated to become
○ 1st block
NADP+
○ 2nd block
○ NADP+ is a coenzyme involved in the
synthesis of lipids
■ is a preparation for the fertilized
oocyte for the formation of the
plasma membranes for the onset of
cleavage, the next stage of
embryogenesis
● Ca2+ release and protein kinase C
○ stimulate protein synthesis, DNA replication,
and cytoplasmic movements of the
morphogenetic material – the later metabolic
responses of the oocyte
● Fast movement of calcium ions
● movement is traced by a fluorescent dye
○ fast spread of Ca2+ ions
- basically similar pathway in mammals EXCEPT the
formation of hyaline layer
- hyaline layer is characteristic to aquatic
mammals
● For mammals:
○ protein kinase C activation → synthesize
proteins needed in cell cycle regulation (e.g.
3.FUSION OF THE GENETIC MATERIALS OF THE
cyclein, histones)
SPERM AND OOCYTE ● Fertilization
● Male pronucleus and female pronucleus ○ a wake up call for oocyte to a fast paced
● Sperm penetrates → depolarization of egg’s plasma action
membrane (1st block) → cortical reaction (2nd block) ■ rapid DNA replication
→ hardening of zona pellucida ■ rapid DNA synthesis of DNA
● initially, sperm head is compact building blocks
● once it is inside the oocyte cytoplasm, it ■ rapid for formation of histones
decondensation (nuclear breakdown) → forming the ■ rapid synthesis of cyclins and cyclin
new male pronucleus dependent kinases
○ metaphase II is lifted off at this point ○ oocyte is fully activated already
○ second polar body is released
● pronuclear fusion a.k.a. amphimixis → forms the
zygote nucleus
○ 2nd polar body is seen meaning fertilization
is complete
● protamines replaced by histones
2
TRANSPORT OF GAMETES & FERTILIZATION IN *correction in figure (lower right text): FEMALE ← MALE
MAMMALS ● capacitation
● Ovary ○ period of conditioning for sperms as they
● ostium (opening of fallopian tube) pass thru the female reproductive tract
○ surrounded by finger-like structures, the ○ removal of glycoprotein coat and other
fimbriae seminal proteins that cover the acrosome
■ captures oocyte when is is first cap
released ○ duration vary from species to species
● fallopian tubes ■ mouse - 1hr.
○ ampulla - site of fertilization ■ rabbit - 6hrs
● uterus ■ humans - 5-8hrs.
○ oocytes takes about 4-5 days to reach the ● how about in vitro fertilization?
uterus from the fallopian tubes ○ no capacitation
○ at day 7-8 it has reached the uterus ○ the collected sperms are kept in a
○ oocyte travels to uterus, regardless of capacitating medium to remove the coat
fertilized or not
TRANSPORT OF THE FERTILIZED OOCYTE
● Day 1 - fertilization
● day 2 - first cleavage
● day 3-4 - 4 cell stage
● day 4 - 8 cell stage
● day 6-7 – late stage blastocyst
● day 8-9 – implantation of the blastocyst
SPERM TRANSPORT IN THE FEMALE
REPRODUCTIVE TRACT
● Insemination
○ process of deposition of sperm in female
reproductive tract
○ upper vaginal canal → cervix → transverse UNION OF GAMETES
the muscular uterus → utero-tubal junction ● In mammals:
→ ampulla of fallopian tube (final side of ○ hyaluronidase is released from the
fertilization) acrosomal cap; digest the corona radiata
○ there is hyperactive motility of sperm in the ■ sperm head penetrates the zona
ampulla in order to penetrate the oocyte pellucida, where the sperm specific
● upper vaginal canal receptors are found
○ site of insemination in humans (and common ● acrosine - proteolytic
mammals) enzyme that helps
● uterus penetration and digestion
○ site of insemination for mice and rats of the ZP
● Barriers in the reproductive tract: ■ sperm head and midpiece
○ natural vaginal acidity penetrates the plasma membrane
■ pH 3.5 → pH 7.2 ○ sperm-specific receptor present in the zona
● due to the natural buffering pellucida
capabilities of semen ○ ZP3 proteins in rodents
○ thick cervical mucus
■ if insemination happens during
ovulation, due to hormonal changes
usually, mucus will be watery
● helps swimming of sperm
○ wide uterus
■ like passing the Pacific ocean for
them
○ utero-tubal junction
■ in common mammals, it is not as
constricting
● unlike with mice and rats
3
ACCOMPLISHMENTS OF FERTILIZATION
● Completion of the second meiotic block
○ metaphase II arrest
● restores normal diploid number of chromosomes
○ 46 chromosomes
● sex of the future embryo is determined
● genetic variation
● metabolic activation of the egg
○ ready for the fast paced cleavage
FERTILIZATION OF AMPHIBIANS
● Progesterone
○ lifts off diplotene block
● at fertilization
○ metaphase II is lifted off
● Ca2+ is released and combines with calmodulin
○ leading to the activation of
calmodulin-dependent kinase
■ which degrades cytostatic factor
(CSF)
● Meiosis II and fertilization is further completed
○ polar bodies are released
TOPIC W/O ADDTNL INFO
SUBTOPIC
● A
○ B
■ C
SUBSUBTOPIC
● A
○ B
■ C
Column head Column head Column head
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