电脉冲与化学激活联合处理对猪卵母细胞孤雌发育的影响

Effects of electrical pulses combined with chemical activation on the parthenogenetic development of porcine oocytes

探讨了猪卵母细胞电激活后间隔不同时间用化学药物处理对猪卵母细胞孤雌发育的影响。结果表明 :1)电激活后间隔 0 ,3,4 ,5和 6h再分别用 6 二甲基氨基嘌呤 ( 6 Dimethylaminopurine ,6 DMAP)、放线菌酮 (cyclohex imide ,CHX)处理 6h ,各组卵裂率、囊胚细胞数差异不显著 (P >0 0 5 ) ,但在 6 DMAP组中对照组囊胚率极显著高于其余各组 (P <0 0 1) ,在CHX组中对照组囊胚率极显著高于间隔 5和 6h组 (P <0 0 1) ,而其余各组之间差异不显著 (P >0 0 5 ) ;2 )电激活后间隔 4h再分别用 6 DMAP ,CHX和细胞松弛素B(CytochalasinB ,CB) + 6 DMAP处理 6h ,各组卵裂率差异不显著 (P >0 0 5 ) ,对照组囊胚率极显著高于其余各组 (P <0 0 1) ,其余各组囊胚率显著性均无差异 (P >0 0 5 )。对照组中无单倍体囊胚 ,二倍体囊胚为 88 2 4 % ,但 6 DMAP ,CHX ,CB + 6 DMAP中产生的囊胚大部分是单倍体 ,且三者之间无显著性差异 (P >0 0 5 )。以上结果说明 ,猪卵母细胞电激活后间隔一段时间再用DMAP和CHX激活 ,激活作用随时间的延长而减弱 ;猪卵母细胞电激活后 4h再用 6 DMAP ,CHX ,CB + 6 DMAP激活 ,产生的囊胚大部分为单倍体 ,该方法适于猪卵母细胞ICSI的激活。

The objective of this study was to determine the effects of chemical treatment after electric activation in different intervals on the parthenogenetic development of porcine oocyte. Porcine oocytes were cultured in vitro for 0 (control), 3, 4, 5 and 6?h after electrical activation, they were then incubated in 6 DMAP/CHX. There were no significant differences ( P >0 05) in the mean numbers of nuclei per blastocysts and cleavage rate. But the blastocysts rate of the control group with 6 DMAP was significantly higher than other groups ( P <0 05). Blastocyst rate of the control group with CHX was higher than the groups with 5 and 6?h intervals, ( P <0 05). In the parallel study, there were no significant differences in the cleavage rates ( P >0 05), but the blastocyst rate of the control group was higher than other groups ( P <0 05) The blastocysts cultured in control almost were diploids, while most blastocysts in other groups were haploids, and there was no significant difference ( P >0 05) . These results demonstrate that the activatiing effect gradually declines as the interval time gets longer, and most blastocystes activated by electricity and 6 DMAP, CHX, CB+6 DMAP (with 4?h interval between them) mostly are haploid.