| The process that sperm and oocyte integreated with each other was called by in vitro fertilization(IVF); IVF was the technology that sperm and oocyte were cultivated under the appropriate condition in vitro; It was generally speaking that IVF include in vitro maturation,in vitro fertilization and in vitro cultivation. It was usually that the polyspermy occur in porcine. The chrosome of polyspermic embryo was triple.The development of zygote was hampered after IVF.the majority of scholar studyed from the aspect of the extent of oocyte's maturation,the abnormity of zona pellucida,follicular secretion and oviduct secretion. And these aspects come from the aspect of female.Although some scholars achieved some achievements.they couldn't solve the problem ultimately. In fact, the fertilization was the result that sperm and oocyte interact each other.In vitro capacitation include decapacitation, when to capacitate,how fast to capacitate, the extent of capacitation,acrosome structure and condition.these facts relate to polyspermy. According to the fact of production.The text aimed at studying the influence of the source of semen and whether or not freeze.the author used ejaculated semen, frozen-thawed ejaculated semen,epididymis semen and frozen-thawed epididymis semen as material.Then four kinds of semen were fertilized with oocyte maturated in vitro. The author observed motility, acrosome integrity, male pronucleus formation, polyspermic penetration rate and clevage rate of embryo.the author studyed some factors affecting IVF and the reason of polyspermy from the aspect of male.these results afforded the proof to solve the problem of polyspermy.In accordance with analysis of the result of study,the conclusion was followed by:1,The author compared the motility and the rate of acrosome integrity from four kinds of semen:the author used four kinds of semen as material and fertilized with oocytes.The author detected motility and the rate of acrosome integrity.the study discuss the change of ejaculated semen and epididymis semen suffered from frozen-thawed.the result of study indicated that the motility of frozen-thawed epididymis semen(65.2%,2.6×10~4/4×10~4) was similar to the motility of epididymis semen(82.3%,3.3×10~4/4×10~4) (p＞0.05).The rate of acrosome integrity of frozen-thawed epididymis semen(43.2%, 1.7×10~4/4×10~4) was lower than the rate of acrosome integrity of epididymis semen(62.3%,2.5×10~4/4×10~4) (p＞0.05). the motility of frozen-thawed ejaculate semen(45.5,1.8×10~4/4×10~4) was lower than the motility of ejaculate semen(85.0%,3.4×10~4/4×10~4) (p＜0.05).The rate of acrosome integrity of frozen-thawed ejaculate semen(40.8%,1.6×10~4 /4×10~4) was lower than the rate of acrosome integrity of ejaculate semen(66.4%,2.6×10~4/4×10~4) (p＜0.05).The epididymis semen was lower sensitive to cold and conserved more easily.The membrane of frozen-thawed semen acrosome was less destrory.2,The difference of IVF among four kinds of semen:the author make semen capacitate separately and adjusted the semen concentration to 10~6.The author used four kinds of semen fertilized with oocytes in vitro.after 8～10h(mTBM).the author transfer the zygote to the IVC-Pyrlac-CM.The IVC-Pyrlac-CM was prequilibrated 12h.The zygote was cultivated 48h in IVC-Pyrlac-CM.and the author statisticed the clevage rate.the result of study indicated that the clevage rate from ejaculated semen(43.7%,34/78) was similar to the clevage rate from epididymis semen(42.3%,33/80) (p＞0.05).The clevage rate of frozen-thawed epididymis semen(40.5%) was higher than the clevage rate of frozen-thawed ejaculated semen(13.5%) (p＜0.05). the clevage rate of ejaculated semen(43.7%,34/78) was lower than the clevage rate of frozen-thawed epididymis semen(40.5%,34/84) (p＞0.05),The ability of in vitro fertilization of frozen-thawed epididymis was similar to the ability of in vitro fertilization of ejaculate semen.3,The difference of polyspermy among four kinds of semen in porcine: the author make semen capacitate separately and adjusted the semen concentration to 10~6.The author used four kinds of semen fertilized with oocytes in vitro.after 8～10h(mTBM).The author observed zygote penetrated by semen and statisticed polyspermic penetration rate.after 18h.The zygote was fixed and dyed with H33342.the male pronucleus fromation was observed.the author statisticed the monospermic penetration rate.The result of study indicated that the monospermic penetration rate of frozen-thawed epididymis semen(7.2%,6/84) was lower than the monospermic penetration rate of ejaculated semen(35.5%,27/78) (p＜0.05).The polyspermic penetration rate of frozen-thawed epididymis semen(70.2%,59/84) was higher than the polyspermic penetration rate of ejaculated semen(46.2%,36/78)(p＜0.05). the monospermic penetration rate of epididymis semen(28.5%,22/80) was lower than the monospermic penetration rate of ejaculated semen(35.5%,27/78) (p＞0.05 ).The polyspermic penetration rate of epididymis semen(51.8%,41/80) was higher than the polyspermic penetration rate of ejaculated semen(46.2%,36/78) (p＞0.05 ). The monospermic penetration rate of frozen-thawed epididymis semen(7.2%,6/84) was higher than the monospermic penetration rate of frozen-thawed ejaculate semen (5.2%, 4/78) (p＞0.05).The polyspermic penetration rate of frozen-thawed epididymis semen(70.2%,59/84) was higher than the polyspermic penetration(65.7, 51/78) rate of frozen-thawed ejaculate semen (p＞0.05).In conclusion:the motility of the frozen-thawed epididymis semen,epididymis semen,the frozen-thawed ejaculated semen and the ejaculated semen was 65.2%,82.3%,45.5%,85.0%.The rate of acrosome integrity of the frozen-thawed epididymis semen,epididymis semen,the frozen-thawed ejaculated semen and the ejaculated semen was 43.2%,62.3%,40.8%,66.4%.The result of study indicated that the frozen-thawed epididymis semen was lower sensitive to cold.The membrane of the acrosome was destrory seriously.The author detected the clevage rate of four kinds of semen.the clevage rate of the frozen-thawed epididymis semen, epididymis semen,the frozen-thawed ejaculated semen and the ejaculated semen was 40.5%,42.3%,13.5%,43.7%.The epididymis semen has the ability of nomal fertilization.and the clevage rate of epididymis semen was similar to the clevage rate of ejaculated semen (p＞0.05).The monospermic penetration rate of frozen-thawed epididymis semen was 7.2%.The monospermic penetration rate of frozen-thawed ejaculate semen was 5.2%(4/78),So the monospermic penetration rate of frozen-thawed ejaculate semen was lower than the monospermic penetration rate of frozen-thawed epididymis semen (p＞0.05).The rate of acrosome integrity of unfrozen ejaculate semen and epididymis semen were higher.The polyspermic penetration rate of these kinds of semen were lower.
|
PDF Full Text Request