The Effect Of The Bovine Oocytes Maturating In Vitro And The Activating Treatment To Parthenogenesis

The effects of preserving ovary temperature, exogenous hormone(LH,17β-E2), and various culturing time (22-25h,26-30h, and 31-34h)to the bovine oocytes maturating in vitro were analyzed in this research . The outcome suggested that , the effects of bovine ovary which maturated in vitro preserving temperature(25-29℃,30-35℃, and 36-40℃) to oocytes were not very clear(P>0.05),after statistically analyzed, and the bovine oocytes maturating rates were 55.3%,70.1%,and 60.7%,respectively;the maturity rates of the bundles which were not added hormone, added 17β-E2 , and added LH+17β-E2 were 57.8%,64.3% and 70.1% respectively, and the difference between the bundle which was added LH+17β-E2 and the bundle not added hormone was significance(P<0.05),after statistically analyzed; the mature rate of bovine oocytes which had been cultured in vitro for 31-34h was 68.4%, which was higher than the two bundles which had been cultured for 22-26h(56.3%) and 27-30h (65.4%) respectively, but the differences among the three bundles were not significance(P>0.05),after statistically analyzed. Bovine oocytes which were in maturation mephase (MⅡ)were treated by various concentration of ethanol (5%, 7%, 9%, 11%, and 13%) or calcium ionophore A23187 (2.5μM , 5μM, and 7.5μM)in this research , and observed its activating rate. The best concentrations of ethanol and A23781, which were required to activate ova ,were screened . The bovine oocytes were treated by the compound activity of cycloheximide , and observed their activity rates and the ability of parthenogenesis.The outcome suggested that , the bundles which contained the ethanol 7%, and 9% had differences with other bundles ,and the difference between the two bundles was not significance (P>0.05)after statistically analyzed .But the activity rate (53.1%) of the bundle which contained 9% ethanol was higher than that of the bundle which contained 7% ethanol , so the bundle which contained 9% ethanol was chosen in next step . The difference between the two bundles whose calcium ionophore A23187 concentrations are 5μM or 7.5μM was not significance (P>0.05), but the difference between the two bundles and the bundles whose concentration was 2.5μM was significance (P<0.05)after statistically analyzed, and the activity rate (51.8%) of the bundle whose concentration was 5μM was chosen in next step. Not only was the difference between the single activating treatment and compound activating treatment which contained 9% ethanol with cycloheximide or 5μM calcium ionophore significance, but also the activating rates (77.3%, 78.8% ,respectively) of compound treatment was higher than that of single treatment (53.1%, 51.8% ,respectively), and the difference between the formation rates of 2PN which were treated by single activating treatment and that of the 2PN treated by compound activating treatment was significance (P<0.05)(40.2%,72.9% respectively) after statistically analyzed. The ova which had been disposed by 9% ethanol with cycloheximide were compared in the ability of parthenogenesis ,and the embryos which were developed to 2-8 cells were 71.0% , 62.9% respectively, but the difference between two ways was not significance (P>0.05),after statistically analyzed.