绵羊去透明带卵母细胞的孤雌激活

为探索一种简化的绵羊核移植操作程序,比较了不同浓度离子霉素、两种不同激活方法对保留和去除透明带的绵羊卵母细胞孤雌发育的影响。结果,2.5μmol/L离子霉素+2mmol/L 6-DMAP激活去除透明带卵母细胞时,卵裂率为78.9%,囊胚发育率为13.7%;采用5μmol/L离子霉素+2mmol/L 6-DMAP方法激活保留透明带的卵母细胞时,卵裂率为88.5%,囊胚发育率为14.7%。在WOWs培养中,对去除和保留透明带的卵母细胞,用5μmol/L离子霉素+2mmol/L 6-DMAP激活时,卵裂率分别为70.4%和69.2%,用70mL/L无水乙醇+2mmol/L 6-DMAP激活时,卵裂率分别为33.3%和23.0%,离子霉素处理组的卵裂率显著高于无水乙醇处理组。结果表明,透明带的去除与否对卵裂率没有产生影响,但各处理组的卵裂卵均未发育为囊胚。

Parthenogenetic activation of zona-free ovine oocytes

In this research the effects of two activation methods and different doses of activators on zona-intact and zona-free oocytes parthenogenetic development were compared to establish a simple nuclear transfer procedure. Zona-free oocytes were treated with 2.5μmol/L ionomycin and 2mmol/L 6-DMAP for the parthenogenetic activation,respectively,the rate of cleavage was 78.9%,and the development rate to blastocysts was 13.7%.Zona-intact oocytes were treated with 5μmol/L ionomycin and 2mmol/L 6-DMAP,respectively,and the rate of cleavage was 88.5% and the development rate to blastocysts was 14.7%.Zona-intact and zona-free oocytes cultured in WOWs were treated with 5μmol/L ionomycin and 6-DMAP for parthenogenetic activation,and the rates of cleavage were 70.4% and 69.2%,respectively.Zona-intact and zona-free oocytes were treated with 7% ethanol and 6-DMAP for parthenogenetic activation,and the rates of cleavage were 33.3% and 23.0%,respectively.The cleavage rates of oocytes treated with ionomycin were significant high than that with 7% ethanol.The results indicated that cleavage rate was not affected by zona-intact or zona-free of oocytes,but the cleavage oocytes cultured in WOWs did not develop to blastocyst.