两种激活处理促进小鼠孤雌胚胎原核形成

不同人工处理方法激活哺乳动物卵母细胞的机理相似,但其激活效率存在差异。本研究以昆明（KM）、129/Sv×KM F1和C3H×KM F1雌鼠来源的卵母细胞为对象,利用氯化锶（SrCl2,Sr2＋）联合细胞松弛素B（cytochalasin B,CB）（Sr2＋＋CB）和离子霉素（ionomycin,Ion）联合6-二甲胺基嘌呤（6-dimethylaminopurine,6-DMAP）（Ion＋6-DMAP）两种激活方法处理下对比分析不同品系小鼠卵母细胞的激活效率,并以卵母细胞原核形成率、原核数量和孤雌胚胎体外发育来评价两种激活剂的激活效率。研究结果表明,Ion＋6-DMAP激活卵的1原核比率显著高于2原核（p〈0.05）,Sr2＋＋CB激活卵的2原核比率显著高于1原核（p〈0.05）;KM、129/Sv×KM F1和C3H×KM F1各组孤雌胚胎卵裂率和激活率没有显著差异（P〉0.05）,但129/Sv×KM F1和C3H×KM F1囊胚发育率显著高于KM组（p〈0.05）。3种小鼠品系的卵母细胞用Sr2＋＋CB处理的孤雌胚胎发育率显著高于Ion＋6-DMAP。结果证明,Sr2＋＋CB处理小鼠卵母细胞的激活效率明显优于Ion＋6-DMAP;129/Sv×KM F1和C3H×KM F1的孤雌胚胎体外发育率显著高于KM小鼠,为研究小鼠遗传背景影响孤雌胚胎发育的机理提供参考。

Two Activation Methods Promote Pronuclear Formation of Parthenogenetic Embryos in Mice

The activation mechanism is similar for mammal oocyte with artificial activation methods,by which the activation efficiency was different.To explored the relations of activate methods on activation efficiency of oocyte,oocyte derived from Kunming（KM）,129/Sv×KM F1 and C3H×KM F1 was used as research objects to investigate the activation efficiency.The two methods were that SrCl2（Sr2＋） was combining with cytochalasin B（CB）（Sr2＋＋CB） and ionomycin（Ion） was combining with 6-dimethylaminopurine（6-DMAP）（Ion＋6-DMAP）,respectively.Activation efficiency was evaluated by the formation rate and number of pronuclear and in vitro development of parthenogenetic embryo.The results showed that the ratio of one pronuclear in 1-cell stage and the fragment rate of cytoplasm in 2-cell stage for Ion＋6-DMAP was higher than that of Sr2＋＋CB（p0.05）;the ratio of the two pronuclear in 1-cell stage for Sr2＋＋CB was higher than Ion＋6-DMAP（p0.05）.For the two methods,the efficiency of activation and cleavage were no significant different（p0.05）,but the rate of blastocyst development for 129/Sv×KM F1 and C3H×KM F1 was more than that of KM（p0.05）.For each strain,the rate of blastocyst development for Sr2＋＋CB was more than that of Ion＋6-DMAP（p0.05）.The results demonstrated that the activation effect of Sr2＋＋CB was better than Ion＋6-DMAP obviously;the development rate of 129/Sv×KM F1 and C3H×KM F1 parthenogenetic embryo was higher than that of KM mice,which provides reference for the research of genetic background on parthenogenetic embryos in mice.