| 摘要: |
| 比较了Ionomycin(离子霉素)和乙醇及其与其他物质的组合激活牛卵母细胞的效果，并研究了去核时间、细胞松弛素B、末期去核等对去核效率的影响。结果表明，配合使用6-二甲氨基嘌呤(6-DMAP)或放线菌酮(CHX)可显著提高Ionomycin或乙醇激活牛卵母细胞的效率，Ionomycin +6-DMAP激活的牛孤雌胚在8/16细胞之后的发育率明显高于乙醇 +6-DMAP(或CHX)。牛卵母细胞的去核在成熟培养20 h或之前进行效果较好，此时有较高比率的卵母细胞中第一极体与染色体位置靠近。牛卵母细胞在不含细胞松弛素B的操作液中可进行去核，但所得核移植胚胎的卵裂率明显下降，而对8/16细胞胚之后的发育没有明显影响。牛卵母细胞的末期去核率显著高于中期，二者核移植胚胎的早期发育率基本相同。 |
| 关键词: 卵母细胞 牛 去核 激活 |
| DOI： |
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| 基金项目:国家自然科学基金项目(39830280) |
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| Bovine oocytes enucleation and artificial activation |
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| Abstract: |
| Activation efficiency of ionomycin,ethanol and their association with 6-DMAP or CHX on bovine oocytes was compared in the present experiment.Telophase (TⅡ) bovine oocytes enucleation,the effect of IVM time and cytochalasin B (CB) on metaphase (MⅡ) enucleation were also studied.The results indicated that:The activation efficiency of bovine oocytes by Ionomycin+6-DMAP or ethanol+6-DMAP/CHX was significantly higher than Ionomycin or ethanol alone.The parthenogenetic development was more stable when bovine oocytes were activated by Inonomycin + 6-DMAP as compared with ethanol+6 DMAP/CHX,especially from 8/16 cells stage to blastocyst.The distance of the first polar body from the chromosome became larger when oocytes in vitro maturation culture time increased.Bovine oocytes should be enucleated at/before IVM 20 h.The TⅡ enucleation rate was significantly higher than MⅡ enucleation;their nuclear transfer embryos had about same development ability.The enucleation manipulation could complete when bovine oocytes placed in media without CB,but the presence of CB in enucleation manipulation media could protect oocytes,the function was mainly exhibited in the early cleavage before 8/16 cells stage. |
| Key words: oocyte bovine enucleation activation |
