L06: Fertilisation Flashcards
How does cumulus-oocyte complex enter + move along uterine tubes?
Follicular fluid contains chemoattractants that attract fimbrae to ovulation site
Muscular contractions of tube transports it through
Describe the journey of sperm through female tract in terms of numbers
~4-250 million sperm ejaculated
100,000 make it into uterus
1000 enter each tube via uterotubal junction
10-100 sperm actually reach egg
Describe the journey of sperm through the female tract
Seminal plasma is alkaline so buffers acidic vagina, PSA from prostate causes clotting of semen to prevent flowback
Other enzymes cause liquefaction after 1hr
Sperm must get through cervical mucus to enter uterus - least viscous around ovulation
Contractions of myometrium propel sperm towards tubes
Stem enters tube through uterotubal junction
Chemotaxis (maybe P4) attracts more sperm towards tube with egg in
Muscular actions of tube move sperm towards egg
Where in the female tract does fertilisation commonly occur?
Ampulla
What changes occur during ovulation that make it easier for sperm to get to egg?
Thinning of cervical mucus (day 9-26), effect of oestrogen
Contraction frequency of myometrium increases
Chemotaxis towards correct tube
What are the obstacles faced by sperm in female tract?
- Uterine cavity is v small
- Has to squeeze through muscular walls
- Folded lumen of uterine tubes cause sperm to get stuck
- Viscous fluid within tubes
What is the window of fertilisation determined by?
Spermatogenesis viable for 5 days, egg only for 24h
Hence fertilisation can occur in ~6 days
How does sperm interact with egg?
- Penetration of cumulus
- Binding to ZP + acrosome reaction
- Penetration of ZP and fusion to egg
How does sperm penetrate cumulus cells?
Cumulus cells embedded in hyaluronic acid matrix (holds cells together)
Sperm has hyaluronidase enzyme that breaks down + disperses cumulus
How does sperm penetrate ZP + fuse w egg
Hyper motility of sperm allows it to get through ZP after acrosome reaction
Sperm enters perivitelline space + equatorial segment of sperm head fuses w egg
Interaction if izumo protein + juno receptor
Egg plasma membrane engulfs sperm into vesicle
Causes large influx of Ca2+ from site of sperm entry
What occurs after sperm fuses w egg?
Influx of Ca2+ ACTIVATES oocyte
PLC zeta then leads to Ca oscillations every 3-15 mins for several hrs
What does activation of egg lead to?
- Release of meiotic block
2. block to polyspermy
How does block to polyspermy occur?
Ca influx —> depolarisation which inhibits further fusion
Cortical granules release protease —> zona reaction
- hardening of ZP preventing penetration
- cleavage of ZP2 (key for sperm binding)
Junk receptor lost from egg
What occurs once sperm in inside egg?
Sperm DNA decondenses
Make + female pronuclei move towards eachother, guided by sperm aster (forms from centriole)
Formation of spindles from centrosome
What is syngamy?
Male + female genetic material combine Pro nuclear membranes break down Chromatic intermixes New nuclear envelope forms End of fertilisation + start of embryogenesis = end of syngamy + beginning of cleavage
What day does implantation occur?
Day 6/7
What day does zygote usually enter uterus
Day 4, aided by relaxed musculature due to progesterone
What is a major instance zygote has to cross to enter uterus?
Isthmic sphincter of uterotubal junction - this is relaxed by progesterone
What occurs during cleavage?
Zygote divides until morula stage (16 cells), no formation of cytoplasm hence overall size stays the same, each cell gets smaller
What is the significant about the 8 cell stage of cleavage?
- embryo transfer most commonly occurs for IVF (embryonic genome is activated)
- genetic testing can be done pre implantation (PCR/FISH)
When does the embryonic genome get activated?
4-8 cells stage