不同化学激活方法对小鼠卵母细胞孤雌激活效果的影响

Effects of Different Chemical Activations on the Development of Mouse Parthenogenetic Embryos

  • 摘要: 通过比较几种常用的化学激活方法对小鼠卵母细胞的孤雌激活和孤雌生殖胚胎体外发育的影响,从而筛选出小鼠卵母细胞孤雌激活的适宜方法.选取成熟小鼠卵母细胞,随机分为以下9组进行激活处理:① 离子霉素 (I) + 6-二甲氨基嘌呤 (D);② I+ D+细胞松弛素B (CB);③ I+放线菌酮 (C);④ I+ C+ CB;⑤ 7% 乙醇 (E) + D;⑥ E+ D+ CB;⑦ E+ C;⑧ E+ C+ CB;⑨ Sr2++ CB.比较了这9种化学激活方法以及氯化锶的作用时间和卵龄的不同对对小鼠卵母细胞激活的影响.实验结果表明:小鼠卵母细胞孤雌激活率与激活措施和卵龄有关;小鼠卵母细胞可以被I+D+CB或SrCl2+CB有效激活,激活率及孵化囊胚发育率均高于其他激活方法(P0.05),且以SrCl2+CB最为有效.hCG注射后20h 为小鼠卵母细胞进行SrCl2 激活的最适卵龄,其最佳处理时间为4h .对小鼠卵母细胞而言,6-DMAP比放线菌酮的激活效果好.

     

    Abstract: In order to find out the suitable activation protocol for mouse oocytes, the cleavage and development of mouse oocytes activated by several different chemical activation protocols were investigated. Matured oocytes were grouped into 9 treatments randomly: (1) inomycin + 6-DMAP; (2) ionomycin + 6-DMAP+ cytochalasin B (CCB); (3) ionomycin+ cycloheximide (CHX); (4) ionomycin + CHX + CCB; (5) 7% ethanol + 6-DMAP; (6) 7% ethanol + 6-DMAP + CCB; (7) 7% ethanol + CHX; (8) 7% ethanol + CHX + CCB; (9) Sr2+ + CCB. The parthenogenetic efficiencies of these activation protocols on mouse oocytes, the optimum treatment duration and the age of oocytes activated by Sr2+ + CCB were analysed. The resultus indicated that the parthenogenetic efficiencies of mouse oocytes are related with activation protocols and the age of oocytes. Mouse oocytes can be activated effectively by ionomycin + 6-DMAP + cytochalasin B or Sr2+ + CCB, the rate of activated oocytes and the development rate of hatched blastocysts in these groups are higher than that in other treatment groups (P0.05). Sr2+ + CCB protocol is the best method of mouse oocyte activation. Mouse oocytes at 20h post hCG injection are the optimum stage for parthenogenetic activation by Sr2+ + CCB and 4h is the best treatment duration for Sr2+ + CCB activation. 6-DMAP is more effective than CHX in activating mouse oocytes.

     

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