| 摘要: |
| 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)；②I + D + 细胞松弛素B(CCB)(IDC) ；③I + 放线菌酮(X)(IX)；④I + 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5 min,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51.0%；培养到196 h时，各组的囊胚发育率分别为18.2%，24.5%，7.2%,11.9%，11.1%，19.4%，6.7%，12.2%和2.0%。结果表明,在10 mmol/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-MAP +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。 |
| 关键词: 卵母细胞 孤雌激活 化学激活 激活方法 牛 |
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| 基金项目:国家自然科学基金资助项目(39830280) |
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| Effect of different chemical activations on the development of bovine parthenogenetic embryos |
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| Abstract: |
| 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. |
| Key words: oocytes parthenogenesis chemical activation activation procedure bovine |
