| The experiment studied in four areas, including factors affecting rabbitsuper-ovulation, oocytes matured in vitro system, parthenogenetic activation ofoocytes and parthenogenetic embryos cultured in vitro. Thus optimizing the systermof rabbit oocytes culture in vitro, parthenogenetic activation and parthenogeneticembryos culture, the foundation is laid by the establishment of in vitro oocytesdevelopment technology platform for the study of rabbit oocytes and parthenogenesisrelated mechanism. Getting the following results:1The test studied the effects of rabbits months of age, hormone dosage andseason three factors on rabbit superovulation. The results indicate that, superovulated4,6,8,12-month-old females respectively, There was no significant (P>0.05) effecton different months of age in the average number of eggs ovulated. This test selected4-month-old females performed superovulation. Using the combination of PMSG andhCG for4months old females superovulation, the average number of oocytesobtained in80IU was significantly (P <0.05) higher than60IU and100IU. Thenumber of eggs ovulated was significantly (P <0.05)higher in spring than both in thewinter and summer, as much as (94.7±2.51) pieces, The rates of oocyte maturationand parthenogenetic activation in spring were significantly (P <0.05) higher insummer and winter; autumn came second. But the difference was not significant.2The test studied the influence of the maturation of rabbit oocytes in vitro todifferent types of oocytes and adding different concentrations of PMSG in vitroculture system. The results indicate that,4different types of oocytes were cultured invitro maturation. Oocyte of grade A and B had a significantly(P<0.05) highermaturation rate. Rabbit oocytes were matured in vitro by adding different concentrations of PMSG (0,0.5,1,1.5,2IU/mL) in the maturation medium. Theoocyte maturation rate of the1IU/mL PMSG was the highest, as much as70.08%,significantly(P <0.05) higher than all other treatment groups.3Comparison of different methods on parthenogenetic activation of matureoocytes activated. The results indicate that, the rabbit oocytes could be activated byionomycin treated for5min and by ethanol for7min and the cleavage rates ofionomycin-treated group were higher than that of ethanol-treated group. But bothgroups all had low blastula rates, and there was no significant(P>0.05) differencebetween them. Using the combination of6-DMAP+ionomycin, the cleavage rate ofionomcine and6-DMAP group was significantly(P<0.05) higher than that ofethanol+6-DMAP group. With the combination of ionomycin, the group of5.0htreatment with2.0mmol/L6-DMAP got the highest cleavage (75.6%) and blastocystformation rate (45.5%).4Comparison of the effect of different medium and culture methods forparthenogenetic embryos in vitro. The results showed that, rabbit parthenogeneticembryos were cultured in vitro in three different medias (M-199, DMEM andcleavage solution), the cleavage rate and blastocyst rate of parthenogenetic embryowhich were cultured in liquid group were significantly(P <0.05) higher than the othertwo group. The rate of parthenogenetic embryo cleavage in the cleavage culturemedium adding0.1%mercaptoethanol was significantly(P <0.05) higher, but the rateof blastocyst was lower (P <0.05). The parthenogenetic embryos were placed incleavage liquid adding0.1%mercaptoethanol for12h, then transferred to thecleavage liquid medium, the culture medium was changed every other day, thecleavage and blastocyst rate of parthenogenetic embryos increased significantly, asmuch as89.55%and47.58%. |
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