Zona Pellucida

glycoprotein shell ZP1, ZP2, ZP3, ZP4 (mouse only ZP1-3)

mechanical protection of egg

involved in the fertilization process

sperm binding

adhesion of sperm to egg

acrosome reaction

releases enzymes to locally breakdown

block of polyspermy

altered to prevent more than 1 sperm penetrating

may also have a role in development of the blastocyst

Fertilization Site

Fertilization usually occurs in first 1/3 of oviduct

Fertilization can also occur outside oviduct, associated with In Vitro Fertilization (IVF, GIFT,
ZIFT...) and ectopic pregnancy
The majority of fertilized eggs do not go on to form an embryo

Fertilization - Spermatozoa

Sperm Binding - zona pellucida protein ZP2 acts as receptor for sperm
Acrosome Reaction - exyocytosis of acrosome contents (Calcium mediated) MBoC - Figure
20-31. The acrosome reaction that occurs when a mammalian sperm fertilizes an egg
enzymes to digest the zona pellucida

exposes sperm surface proteins to bind ZP2

Membrane Fusion - between sperm and egg, allows sperm nuclei passage into egg
cytoplasm

Contact between spermatozoa and oocyte egg coat (zona pellucida [ZP]) glycoproteins
triggers increases in intracellular calcium ion (iCa2+) concentration in spermatozoa[8]

CATSPER channels on the distal portion of sperm (the principal piece) are required for the
ZP-induced iCa2+increases

iCa2+ increase starts from the spermatozoa tail and propagates toward the head

Store depletion-activated Ca2+ entry is thought to mediate the sustained phase

Fertilization - Oocyte

Membrane Depolarization - in non-mammalian species, caused by sperm membrane

fusion, acts as a primary block to polyspermy.

Cortical Reaction - IP3 pathway elevates intracellular Calcium, exocytosis of cortical

granules

enzyme alters ZP2 so it will no longer bind sperm plasma membrane

Meiosis 2 - completion of 2nd meiotic division
forms second polar body (a third polar body may be formed by meiotic division of the
first polar body)

Spermatozoa - Oocyte Interaction

Membrane Fusion
Spermatozoa and oocyte fusion in the membrane adhesion area requires the presence of 3
membrane proteins (spermatozoa Izumo1; oocyte receptor Juno and Cd9).[9]

Izumo
The sperm-specific protein Izumo is named for a Japanese shrine dedicated to marriage and is
essential for sperm-egg plasma membrane binding and fusion. It interacts with the spermatozoa
folate receptor 4 (Folr4).

Juno
folate receptor 4 (Folr4)
Folate receptors are also known as the folic acid (FA) binding protein and bind of 5methyltetrahydrofolate (5-MeTHF).

CD9
Oocyte cell surface protein acts as a receptor for pregnancy-specific glycoprotein 17 (Psg17).

Ovastacin
(Astacin-Like Metalloendopeptidase, ASTL) An oocyte enzyme (zinc-dependent metalloprotease)
involved in altering zona pellucida structure, by cleaving zona pellucida glycoprotein (ZP2), following
fertilisation.[10] This change in zona pellucida structure is one of the blockers to polyspermy.

Formation of the Zygote

Early human zygote showing Pronuclei

Pronuclei - Male and Female haploid nuclei approach each other and nuclear membranes
break down

chromosomal pairing, DNA replicates, first mitotic division

Spermatozoa contributes - centriole which organizes mitotic spindle

Oocyte contributes - mitochondria (maternally inherited)

Mitochondria

Mitochondria of the spermatozoa are specifically destroyed in early development by

proteolysis (mouse 4 to 8 cell transition).[11]

Metaphase II oocytes in rats have an average mitochondrial DNA (mtDNA) copy number of
147,600 (+/-3000) that only increases at the 8-cell stage.[12]

Sex Determination

based upon whether an X or Y carrying sperm has fertilized the egg, should be 1.0 sex ratio.

actually 1.05, 105 males for every 100 females, some studies show more males 2+ days
after ovulation.

cell totipotent (equivalent to a stem cell, can form any tissue of the body)
Men - Y Chromosome

Y Chromosome carries Sry gene, protein product activates pathway for male gonad (covered
in genital development)
Women - X Chromosome

Gene dosage, one X chromosome in each female embryo cell has to be inactivated

process is apparently random and therefore 50% of cells have father's X, 50% have mother's
X

Note that because men only have 1 X chromosome, if abnormal, this leads to X-linked
diseases more common in male that female where bothe X's need to be abnormal.