几种不同化学激活方法对牛卵母细胞孤雌激活的影响

为确定牛体细胞核移植胚胎的激活条件 ,比较了几种不同化学激活方法对牛卵母细胞激活的影响。选择明显具有第一极体的成熟卵母细胞 ,随机分为 9组 :1Ionomycin(I) +6 -甲氨基嘌呤 (D) (ID) ;2 I+D+细胞松弛素 B(CCB) (IDC) ;3I+放线菌酮 (X) (IX) ;4 I+X+CCB(IXC) ;5体积分数 7%乙醇 (E) +D(ED) ;6E+D+CCB(EDC) ;7E+X(EX) ;8E+X+CCB(EXC) ;9Sr2 + +CCB(SC)。Ionom ycin和乙醇的处理时间为 5min,6 - DMAP± CCB的处理时间为 4 h,放线菌酮± CCB的处理时间为 5 h,Sr2 + +CCB的处理时间为 10 h。培养36 h后 ,各组卵母细胞的卵裂率分别为 72 .0 % ,75 .5 % ,81.3% ,85 .1% ,81.7% ,82 .4 % ,74 .4 % ,78.1%和 5 1.0 % ;培养到 196 h时 ,各组的囊胚发育率分别为 18.2 % ,2 4 .5 % ,7.2 % ,11.9% ,11.1% ,19.4 % ,6 .7% ,12 .2 %和 2 .0 %。结果表明 ,在 10 mm ol/ L Sr2 +内培养 10 h不能有效地激活牛卵母细胞 ,Ionom ycin+6 - DMAP± CCB以及乙醇 +6 -DMAP+CCB结合使用可以有效地激活牛卵母细胞。Ionom ycin比乙醇对牛卵母细胞的激活作用强 ;6 - DMAP比CHX的激活效果好 ,激活液中 CCB的存在对牛孤雌胚胎的发育有利

Effect of different chemical activations on the development of bovine parthenogenetic embryos

The cleavage and development of bovine oocytes activated by different chemical activative procedures were investigated.Matured oocytes were grouped into 9 treatments randomly:(1) Ionomycin(I)+6-DMAP (ID);(2) Ionomycin+6-DMAP+ cytochalasin B(CCB) (IDC);(3) Ionomycin+Cycloheximide(CHX) (IX);(4) Ionomycin+CHX+CCB(IXC);(5) 7% ethanol+6-DMAP (ED);(6) ethanol+6-DMAP+CCB (EDC);(7) ethanol+CHX (EX);(8) ethanol+CHX+CCB(EXC);(9) Sr²⁺ +CCB(SC).The treatment time for Ionomycin or 7% ethanol,6-DMAP±CCB,CHX±CCB and Sr²⁺ +CCB were 5 min,4 h,5 h and 10 h,respectively.The cleavage rates of each group were 72.0%,75.5%,81.3%,85.1%,81.7%,82.4%,74.4%,78.1% and 51.0% respectively after cultured for 36 h and blastocyst rates were 18.2%,24.5%,7.2%,11.9%,11.1%,19.4%,6.7%,12.2% and 2.0% respectively after cultured for 8 days.These results indicate that bovine oocytes can be activated by ID,IDC and EDC effectively,whereas oocytes cannot be stimulated effectively by 10 mmol/L Sr²⁺ for 10 h.Ionomycin has strong activation effect when compared with ethanol,while 6-DMAP is more effective than cycloheximide in activating bovine oocytes.Cytochalasin B is advantageous for the development of bovine parthenogenetic embryos.