化学激活对猪体外成熟卵母细胞孤雌发育的影响  

Effects of Chemical Activation on the Parthenogenetic Development of Porcine in vitro Maturation Oocytes

作　　者：田见晖[1] 蔡元[2] 刘国世[1] 曾申明[1] 成文敏[1] 朱士恩[1] 刘焕营[1] 吴常信[1] 

机构地区：[1]中国农业大学动物科技学院/农业部畜禽遗传育种重点开放实验室,北京100094 [2]甘肃农业大学动物科技学院,兰州730070

出　　处：《中国农业科学》2005年第5期1029-1033,共5页Scientia Agricultura Sinica

基　　金：北京市科委重点项目(H022020060420);高校博士点基金(20020019031)资助项目资助项目

摘　　要：探索了离子霉素结合细胞松驰素B(cytochalasinB,CB)、放线菌酮(cycloheximide,CHX)以及6-二甲基嘌呤(6-dimethylaminopurine,6-DMAP)等化学物质,对猪卵母细胞激活后发育的影响。试验1,体外成熟卵母细胞分别用15、20、25、30μmol·L-1的离子霉素处理40min或电激活(1.3kV·cm-1,80μs,1次脉冲),结果表明,20μmol·L-1组的激活率(69.93±5.80)%显著高于15μmol·L-1组(P<0.05),但与其它各组差异不显著(P>0.05)。试验2,卵母细胞采用20μmol·L-1离子霉素分别激活10、20、30、40、50min,再用2mmol·L-16-DMAP处理6h,其中,40min组的卵裂率、囊胚率[(72.40±13.02)%、(25.37±11.43)%]较高,但与其它各组差异不显著(P>0.05)。试验3,卵母细胞经离子霉素(20μmol·L-1、40min)激活后,分别用7.5μg·ml-1CB、10μg·ml-1CHX、2mmol·L-16-DMAP、7.5μg·ml-1CB+10μg·ml-1CHX和7.5μg·ml-1CB+2mmol·L-16-DMAP处理6h,2mmol·L-16-DMAP组的激活率、卵裂率和囊胚率[(86.05±4.29)%、(61.77±8.10)%和(21.62±3.31)%]显著高于7.5μg·ml-1CB组(P<0.05)与另外几组差异不显著(P>0.05)。试验4,卵母细胞由离子霉素(20μmol·L-1、40min)激活后,用2mmol·L-16-DMAP分别处理3.5、5.5、7.5h,5.5h组的卵裂率和囊胚率[(66.The objective of this study was to determine the effects of ionomycin combined with cytochalasin B (CB), cycloheximide (CHX), or 6-dimethylaminopurine (6-DMAP) on the activation of porcine oocytes. In experiment 1, in vitro matured oocytes were activated with ionomycin at 15, 20, 25 or 30 μmol·L-1. Activation rates of 20, 25 μmol·L-1 and 30 μmol·L-1 treatments were higher (P<0.05)than that of 15 μmol·L-1 treatment. In experiment 2, in vitro matured oocytes were activated with 20 μmol·L-1 ionomycin at 10,20,30,40 or 50 min and then incubated with 2 mmol·L-1 6-DMAP 6 h.Cleavage rates and blastocyst formation rates after 40 min treatments [(72.40±13.02)%, (25.37±11.43)%]was higher (P>0.05) than that of the either treatments. In experiment 3, matured oocytes were activated with ionomycin and then incubated with 7.5 μg·ml-1 CB,10 μg·ml-1 CHX, 2 mmol·L-1 6-DMAP, 7.5 μg·ml-1 CB +10 μg·ml-1 CHX or 7.5 μg·ml-1 CB +2 mmol·L-1 6-DMAP for 6 h. Activation rates, cleavage rates and blastocyst formation rates of 2 mmol·L-1 6-DMAP treatment [(86.05±4.29)%, (61.77±8.10)% and (21.62±3.31)%] was higher (P<0.05) than that of 7.5μg·ml-1 CB treatment. In experiment 4, matured oocytes were activated with ionomycin and then incubated with 2 mmol·L-1 6-DMAP for 3.5, 5.5 or 7.5 h. Cleavage rates and blastocyst formation rates of 5.5 h treatment [(66.59±14.36)% and (25.40±10.16)%] was higher (P>0.05) than that of other treatments. In conclusion, activation of porcine oocytes appears to be most successful using the combination of ionomycin (20 μmol·L-1, 40 min) followed by 6-DMAP (2 mmol·L-1 , 5.5 h).

关 键 词：化学激活 猪 体外成熟 卵母细胞 孤雌发育 离子霉素 

分 类 号：S828[农业科学—畜牧学]