Fertilization

Fertilization: Penetration of the ovum by a spermatozoon and completed by the fusion of the female and male chromosomes with subsequent formation of male and female pronuclei and expulsion of the second polar body, syngamy leading to cleavage to two blastomeres. Events: 1. Gamete Maturation 2. Sperm-Oocyte Encounter 3. Cumulus Penetration 4. Sperm Attachment 5. Zona Penetration 6. Gamete Fusion 7. Block to Polyspermy 8. Development of Pronuclei 9. Syngamy I. Ovum Maturation 1. Most species are in Metaphase II of the second Meiotic Division at Ovulation 2. Ovum Maturation and meiosis are not completed until after fertization. 3. Ovum Longevity A. Cattle 20-24 hr B. Horse 6-8 hr C. Sheep 16-24 hr D. Swine 8-10 hr II. Sperm Maturation 1. Capacitation A. Final maturation of spermatozoa in the female reproductive tract leading to fertilization. B. Cervix removes seminal plasma which inhibits capacitation. C. Process starts in Uterus. D. Major site--oviduct (isthmus) E. Sperm surface components modified and removed by female genital tract secretions. a. Removal of Cholesterol b. Alteration in glycoaminoglycans (GAGs) c. Changes in ions. F. Seminal plasma reverses process --Decapacitation G. Capacitation leads to acrosome changes that lead to sperm penetration of the ovum. H. Capacitation prevents premature acrosome activation --Acrosome reaction. 2. Acrosome Reaction A. Fusion of the the sperm plasma membrane with the outer acrosomeal membrane. --A false acrosome reaction also occurs during senenscence or degeneration of sperm

B. Fusion and Vesiculation of the acrosome allow for release of hydrolytic enzymes needed for ovum penetration 3. Sperm Lifespan A. Farm Species: Cattle 30-48 hr Horse 72-120 hr Sheep 30-48 hr Swine 34-72 hr B. Highest Conception a. If sperm are in oviduct just prior to ovulation III. Interaction of Sperm and Ovum 1. Sperm-oocyte encounter A. Fertilization requires three critical events: a. Sperm migration between cummulus cells b. Sperm attachment and migration through the zona pellucida c. Fusion of sperm and ovum plasma membranes 2. Cumulus Penetration A. Cumulus stimulates sperm activity (rabbits) a. Oviductal secretions cause hyperactivity too. B. Much if not all of cumulus removed by mechanical means as ovum travels through the oviduct. a. Cattle cumulus absent 3-4 hours after fertilization C.Hydrolytic enzymes let sperm migrate through cumulus a. Hyaluronidase Bull b. Arylsulfatase Boar 3. Sperm Attachment A. Attachment of the sperm head to the zona pellucida is mediated by a receptor a. ZP3 Sperm Receptor b. glycoprotein c. Only sperm with intact acrosome can bind d. "Lock & Key" mechanism starts enzyme release for penetration. 4. Sperm Penetration A. Occurs 5 to 15 minutes after sperm attachment. B. Enzymes released to penetrate zona a. At least 9 enzymes released b. Acrosin --major zona lysin --others act synergistically with acrosin c. Sperm motility also required for penetration d. ZP2 may also help in attachment/penetration 4. Gamete Fusion A. Once the zona is penetrated the sperm interacts with the vitelline membrane in the vitelline space. B. Sperm rotates vitelline membrane within the zona. C. Sperm interacts with dense microvilli of vitelline membrane D. Attachment of sperm occurs in the equitorial segment of the plasma membrane of the sperm head.

E. Sperm-Ovum plasma membranes become intermixed F. Sperm head undergoes decondensation 5. Block to Polyspermy A. Following fertilization, the ovum surface changes to prevent fusion of additional sperm B. Cortical Reaction a. Cortical granules are released into the perivitelline space. b. Results in extensive reorganization of the zona pellucida (and/or vitelline membrane) --Hardening of Zona --Inactivation of ZP3 & ZP2 C. Polyspermy a. Polyploid embryos--Death or Abnormal development b. Happens in 1-2% of mammalian fertilizations c. Increased by delayed insemination --Pigs & Sheep 6. Development of Pronuclei and Syngamy A. Fusion causes 2nd polar body expulsion B. Haploid female pronucleus forms C. Decondensation products of sperm head form male pronucleus in ovum cytoplasm D. Pronuclei migrate to ovum center E. Syngamy occurs--Nuclear envelopes disperse and chromosomes mix F. Chromosomes aggregate in Mitotic Prophase G. Cell Division begins--Diploid