Juvenile Rabbits’ Oocytes IVF And The Expression Of Related Gene In The Early Embryo

Juvenile in vitro embryo transfer (JIVET) is a technique which combines juvenile superovalation, in vitro maturation, in vitro fertilization, in vitro culture and embryo thransfer. It is greatly shortening the livestock breeding generation interval, exploiting the excellent dams’ breeding potential, improving the speed and efficiency of genetic improvement, and providing a rich genetic resource for livestock breeding. In vitro fertilization is an important part of JIVET. It can significantly improve the quality of postembryonic growth and development, and can effectively increase the productivity of JIVET by improving the effect of IVF.In this study, the juvenile New Zealand rabbits were as the main research objects. After superovulation and IVM, different concentrations of antioxidant EGCG were added to study the effects on sperm capaciation and IVF which compared with the effect of adult rabbits adding the same concentrations, aiming at establishing an efficient culture system and improving the effect of juvenile rabbits’ IVF. And then, the expression of OCT-4, GDF-9, Bcl-2in embryo were investigated by semi-quantitative RT-PCR in order to study the difference between the early embryos of juvenile and adult rabbits at molecular level. It could provide a theoretical basis for improving the quality of post-embryonic development. The results were showm as follows.1. The concentrations of0,5,10,15,20and25μM EGCG were added in capaciation cultures and the results were detected by using Coomassie Brilliant Blue Staining after progesterone induced acrosome reaction. The results showed that EGCG had an significant influence on capacitation (P<0.05). Sperm capacitation effect which adding10μmol/L EGCG was significantly higher than the other groups (P<0.05).2. The concentrations of0,5,10,15and20μM EGCG were added in IVF cultures. The results showed that EGCG had an significant effect on improving the IVF rate both for juvenile rabbits and adult rabbits (P<0.05). The fertilization rates which adding5,10,15,,20μM EGCG were significant higher than the fertilization rates which adding0μM for juvenile rabbits(P<0.05). The fertilization rates which adding5,10and15μM EGCG were significant higher than the fertilization rates which adding0and20μM for juvenile rabbits and adult rabbits (P<0.05).And the fertilization rates were no significant difference for juvenile and adult rabbits while adding the same concentrations of EGCG (P>0.05).3. The expressions of OCT-4, GDF-9, and Bcl-2were investigated by semi-quantitative RT-PCR both for juvenile and adult rabbits’ embryos. The results showed that the expressions of OCT-4and GDF-9in juvenile rabbits’ embryos were significantly lower than that of adult rabbits’ embryos (P<0.05), while the expression of Bcl-2gene was lower than that of adult rabbits’embryos, but the difference was not significant (P>0.05).