| 刘京威,倪和民,郭勇,徐杨,鲍文玉,刘云海.2009.化学联合激活可明显提高小鼠孤雌胚胎发育率.动物学杂志,44(4):135-141. |
| 化学联合激活可明显提高小鼠孤雌胚胎发育率 |
| Murine Parthenogenetic Embryonic Development can be Obviously Improved by the Associated Chemical Activation |
| 投稿时间：2008-09-24 修订日期：2009-04-29 |
| DOI： |
| 中文关键词: 小鼠 孤雌激活 乙醇 氯化锶 6-DMAP |
| 英文关键词:Mouse Parthenogenesis Ethanol SrCl2 6-DMAP |
| 基金项目:北京市教育委员会科技发展计划重点项目(No.KZ200510020013);面上项目(No.KM200710020006);2007年度教育部留学人员科技活动择优资助项目(启动类) |
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| 中文摘要: |
| 为探讨一种高效的小鼠卵母细胞孤雌激活的方案,进一步提高孤雌囊胚发育率。用不同浓度的氯化锶及不同作用时间的乙醇,并分别联合6-DMAP对不同卵龄小鼠卵母细胞进行活化,统计小鼠卵母细胞卵裂率和体外发育状况。结果显示,15～16h、18～19h和20～21h卵龄组卵母细胞经6mmol/LSrCl2联合6-DMAP处理后,三组的激活率随卵龄增长而升高,其中20～21h卵龄组显著高于15～16h、18～19h组(P<0.05),激活胚胎的发育率以18～19h时最高;6mmol/L和10mmol/L的SrCl2联合6-DMAP均能有效地激活小鼠卵母细胞,激活率分别为76.4%和83.6%,桑葚胚率分别为50.0%和56.3%;70ml/L乙醇联合6-DMAP以处理7min组获得了较好的激活率和囊胚发育率,分别为77.1%和42.4%,囊胚率均显著高于4min和10min处理组(P<0.05)。6-DMAP与SrCl2或乙醇联合应用可以有效抑制第二极体的排出,提高激活胚的二倍体比率;孤雌囊胚的平均细胞数显著低于正常受精囊胚(P<0.05)。不同激活方案对孤雌活化胚的核型和发育能力的作用差异较大,小鼠卵母细胞孤雌激活率与卵龄和激活方案密切相关,其中以卵龄18～19h,70ml/L乙醇或10mmol/L SrCl2联合6-DMAP的激活效果较好。 |
| 英文摘要: |
| The purpose of this study is to establish an efficient way for mouse parthenogenetic activation.Oocytes were recovered from mice at different hours after hCG administration and were denuded of cumulus cells with hyaluronidase.The cleavage and blastocyst rates of mouse oocytes activated with different concentrations of strontium chloride,and different durations of ethanol stimulation with or without 6-dimethylamino-purine,were analyzed.The cleavage rate of oocytes collected at 15-16,18-19,20-21 hours were increased after treatment with 6 mmol/L SrCl2.The cleavage rate of oocytes collected at 20-21 hour was significantly higher than that of oocytes collected at 18-19 or 20-21 hour（P<0.05）.The blastocyst rate of oocytes collected at 18-19 hour was the highest among different groups.The cleavage rate of oocytes treated with 6 mmol/L and 10 mmol/L SrCl2 were 76.4%,83.6%,and the morula rates were 50.0% and 56.3% respectively.The cleavage rate of oocytes treated with 70 ml/L ethanol for 7 min was 77.1%,and the blastocyst rate was also significantly higher than that of the other two groups（P<0.05）.The combination of 6-DMAP and SrCl2 or ethanol,improved the extrusion of second polar body,and the diploid parthenogenetic embryonic rate.The average number of parthenogenetic blastocyst cells was significantly lower than that of normal blastocysts（P<0.05）.The cleavage of mouse parthenogenetic embryos depends on both the oocyte age and activation protocols.The parthenogenetic activation for oocytes collected at 18-19 hour by treatment with 10 mmol/L SrCl2 or 70 ml/L ethanol combined with 6-DMAP,was the most efficient approach. |
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