不同化学激活方法对山羊卵母细胞激活效果的研究

激活是体细胞核移植技术的关键步骤。本研究对常用化学激活剂乙醇、钙离子载体A2 3187和离子霉素以及添加 6 二甲基氨基嘌呤 (6 Dimethylaminopurine ,6 DMAP)和放线菌酮 (cycloheximide ,CHX)对山羊卵母细胞的激活效果以及不同卵龄的卵母细胞激活后体外发育效果进行了比较。结果表明 :离子霉素处理组激活率显著高于乙醇和A2 3187处理组 (89.6 %vs 6 3.5 % ,6 8.7% ;P <0 .0 5 ) ;离子霉素 +6 -DMAP处理组激活率显著高于离子霉素 +CHX和离子霉素 +CHX +细胞松弛素D处理组 (90 .1%vs 5 8.9% ,6 3.7% ;P <0 .0 1) ;体外成熟 2 4、2 6、2 8h组桑椹胚率分别为 16 .0 %、2 0 .6 %和 33.3% ,30h组桑椹胚率下降 (13.8% )。以上结果说明 ,在本研究的激活条件下 ,山羊卵母细胞适宜的激活方法为 :成熟起始后 2 8h使用离子霉素和 6 DMAP进行联合激活。

Effects of chemical methods on activation of goat oocytes

Activation is a key step of somatic cell cloning. In this study, the effects of several chemical activation reagents such as ethanol, calcium ionophore A23187 and ionomycine followed by either phosphorylation inhibitor 6-dimethylaminopurine (6-DMAP) or cycloheximide (CHX) on activation of goat oocytes were compared., In addition ,the development ability of aging oocytes after activation was also studied . The results show: 1)ionomycine group had higher activation rate than ethanol and A23187 groups (89.6% vs 63.5%,68.7%; P<0.05). 2)the activation rate of ionomycine + 6-DMAP group was significantly higher than ionomycine + CHX or ionomycine + CHX + cytochalasin D group(90.1% vs 58.9%,63.7%; P<0.01). 3)the morula rates of 24,26,28 hour groups were 16.0%, 20.6% and 33.3% respectively, while decreased in 30 hours(13.8%). These results suggest that ionomycine combined with 6-DMAP at 28 hour after maturation is the optimal condition for goat oocytes activation.