不同激活方法对绵羊体外成熟卵母细胞孤雌生殖的影响  

THE EFFECT OF DIFFERENT METHODS OF PARTHENOGENETIC ACTIVATION ON IN VITRO MATURED OVINE OOCYTES

作　　者：张红琴[1] 周欢敏[1] 李宝山[1] 张东[1] 朱琳 

机构地区：[1]内蒙古农业大学动物科学与医学学院,呼和浩特010018 [2]内蒙古生物药品厂,呼和浩特010030

出　　处：《内蒙古农业大学学报（自然科学版）》2008年第1期78-81,共4页Journal of Inner Mongolia Agricultural University(Natural Science Edition)

基　　金：内蒙古人才基金

摘　　要：为确定绵羊体细胞核移植胚胎的激活条件,本实验探讨了不同激活方法对绵羊体外成熟卵母细胞的孤雌激活及其后的发育影响。选择体外成熟绵羊卵母细胞,随机分为3组:①电激活+6-DMAP4h;②I(5μm)5min+6-DMAP4h;③7%乙醇7min+6-DMAP4h,培养48h后,各组卵母细胞的卵裂率分别为60.93%,79.96%,78.28%;培养到7d时,各组的桑椹/囊胚发育率分别为17.03%,42.84%,39.39%。化学激活的卵裂率、桑椹/囊胚率显著高于电激活(p<0.05);乙醇与Ionomycin相比,其卵裂率、桑椹/囊胚率差异不显著,但Ionomycin处理组效果稍好于乙醇组。结果表明,电激活、Ionomycin、7%乙醇都可以有效激活绵羊卵母细胞,并孤雌发育到囊胚,Ionomycin的激活效果最佳;同时也比较了不同电压对绵羊体外成熟卵母细胞激活的影响,以1.300kV/cm-1.350kV/cm,脉冲时间10us,脉冲次数为2次,间隔0.5s效果较好。To obtain simple and effective methods of electrofusion and activation for ovine somatic cloning, in this study, the different experiments of parthenogenetic activation were examined. The selected in vitro matured ovine oocytes, divided into three parts, were tested to record the parthenogenetic activation rates of the 3 different treatments ： ①Electroactivation combined with 2mM 6 - DMAP for 4 hours; ②5μM Inomycin for 5 minutes combined with 2mM 6 - DMAP for 4 hours; ③7% ethanol for 7minutes combined with 2mM 6 - DMAP for 4 hours. The percentages of cell divisions for the three treatments after in vitro cultured 48 hours were 60.93%, 79. 96%, and 78.28% respectively; and the percentage of formation of morula/blastocyst after in vitro cultured 7 days were 17.03%, 42.84%, and 39.39% correspondingly. Following the treatments, the success rates of parthenogenesis of latter two groups were significant higher than the treatment of electroactivation combined with 6 - DMAP. There was no significant difference between latter two groups. However, the treatment of Inomycin combined with 6 - DMAP was slightly better than the treatment of ethanol combined with 6 -DMAP. The results indicate that the in vitro matured ovine oocytes were effectively activated by electroactivation, 5μM lnomycin for 5 minutes, 7% ethanol for 7minutes combined with 2mM 6 -DMAP for 4 hours unanimously, and parthenogeneticly developed to the morula and blastocyst. Also,in this study, the effects of different voltages used in electroactivation on in vitro matured ovine oocytes were compared. The results indicate that the voltage between 1300 - 1350 kV/ cm with duration of 10μs and 2 times of repeating are the best setting.

关 键 词：卵母细胞 孤雌激活 绵羊 

分 类 号：S826.2[农业科学—畜牧学]