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Fertilization

The process Assessment IVF ICSI

The Process - sperm

capacitation hyperactivation zona binding acrosome reaction zona penetration sperm-oocyte membrane fusion sperm engulfed decondensation of sperm nucleus formation of male pronucleus

The Process - Egg

Ovulation Sperm-egg membrane fusion Egg activation increase in Ca++ cortical reaction - zona block completion of meiosis (2nd PB) formation of female pronucleus Syngamy - fusion of both pronuclei

Capacitation



process of sperm becoming fertile occurs in uterus must be removed from seminal fluid in vitro, wash sperm capacitated sperm have the ability to fertilize the egg

Hyperactivation

facilitates sperm-oocyte contact occurs in oviduct motility pattern becomes frenzied linear to non-linear motion

Zona Binding

Zona proteins: ZP1, ZP2, ZP3 Sperm bind ZP3 specifically Initiates acrosome reaction

Acrosome Reaction

Fusion of two membranes spermatazoal plasma membrane outer acrosomal membrane Acrosin released allows sperm to penetrate zona Exposes equatorial segment for fusion with egg

Sperm-Egg Fusion

Equatorial segment of sperm Plasma membrane of egg Initiates egg activation

Egg Activation

Cortical Reaction exocytosis of cortical granules Zona block biochemical changes eliminates sperm binding

Assessment of Fertilization

Check for PN 15-18 hrs post insemination Normal Fertilization 2 PN in ooplasm 2 PB perivitelline space

Abnormal Fertilization



> 2 PN in ooplasm polyspermic: >1 male PN, 1 female PN, 2PB polygynic: >1 female PN, 1 male PN, 1PB failure to extrude second PB Spontaneous oocyte activation 1 female PN, 2 PB Condensed sperm: 1 female PN, 2 PB Oocyte activation (digynic) >1female PN, 1 PB

IVF: Why?

Humans: overcome infertility male factor: low motility, low count female factor: tubal, ovulatory Animals: end of reproductivity more superior animals Research: IVM with IVF many embryos to study techniques: transgenics, es cells, cloning

Intracytoplasmic Sperm Injection

Overcome fertilization problems: no penetration ASAB, anti-sperm antibodies very low count (<500,000/sample) very low motility (<20% motile)

ICSI - process

Add sperm to PVP inert, slows sperm to catch Mature egg w/holding pipet PB at 12:00 or 6:00 Catch sperm and kink tail Load sperm in ICSI pipet Break plasma membrane Deposit sperm






رفعت المحاضرة من قبل: Abdalmalik Abdullateef
المشاهدات: لقد قام 7 أعضاء و 124 زائراً بقراءة هذه المحاضرة








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