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
Sperm motility also required for penetration d. Sperm Lifespan A. Sperm interacts with dense microvilli of vitelline membrane D. Cattle cumulus absent 3-4 hours after fertilization C. ZP2 may also help in attachment/penetration 4. C.
. ZP3 Sperm Receptor b. Enzymes released to penetrate zona a. Cumulus Penetration A. If sperm are in oviduct just prior to ovulation III. B. Fusion and Vesiculation of the acrosome allow for release of hydrolytic enzymes needed for ovum penetration 3. Farm Species: Cattle 30-48 hr Horse 72-120 hr Sheep 30-48 hr Swine 34-72 hr B.Hydrolytic enzymes let sperm migrate through cumulus a. Interaction of Sperm and Ovum 1. Acrosin --major zona lysin --others act synergistically with acrosin c. glycoprotein c. Gamete Fusion A. Sperm attachment and migration through the zona pellucida c. Attachment of the sperm head to the zona pellucida is mediated by a receptor a. Sperm-oocyte encounter A. Attachment of sperm occurs in the equitorial segment of the plasma membrane of the sperm head.B. 4. Sperm rotates vitelline membrane within the zona. "Lock & Key" mechanism starts enzyme release for penetration. Much if not all of cumulus removed by mechanical means as ovum travels through the oviduct. B. Occurs 5 to 15 minutes after sperm attachment. B. Sperm Attachment A. Oviductal secretions cause hyperactivity too. Once the zona is penetrated the sperm interacts with the vitelline membrane in the vitelline space. Highest Conception a. Hyaluronidase Bull b. Only sperm with intact acrosome can bind d. At least 9 enzymes released b. Sperm migration between cummulus cells b. a. Sperm Penetration A. Cumulus stimulates sperm activity (rabbits) a. Fertilization requires three critical events: a. Fusion of sperm and ovum plasma membranes 2. Arylsulfatase Boar 3.
Cortical Reaction a. Pronuclei migrate to ovum center E. the ovum surface changes to prevent fusion of additional sperm B.E. Increased by delayed insemination --Pigs & Sheep 6. Polyspermy a. Fusion causes 2nd polar body expulsion B. Decondensation products of sperm head form male pronucleus in ovum cytoplasm D. Haploid female pronucleus forms C. Block to Polyspermy A. Polyploid embryos--Death or Abnormal development b. Following fertilization. b. Syngamy occurs--Nuclear envelopes disperse and chromosomes mix F. Development of Pronuclei and Syngamy A. Chromosomes aggregate in Mitotic Prophase G. Cortical granules are released into the perivitelline space. Happens in 1-2% of mammalian fertilizations c. Cell Division begins--Diploid
. Results in extensive reorganization of the zona pellucida (and/or vitelline membrane) --Hardening of Zona --Inactivation of ZP3 & ZP2 C. Sperm head undergoes decondensation 5. Sperm-Ovum plasma membranes become intermixed F.