中国农业科学 ›› 2005, Vol. 38 ›› Issue (05): 1029-1033 .

• 畜牧·兽医·资源昆虫 • 上一篇    下一篇

化学激活对猪体外成熟卵母细胞孤雌发育的影响

田见晖,蔡元,刘国世,曾申明,成文敏,朱士恩,刘焕营,吴常信   

  1. 中国农业大学动物科技学院
  • 收稿日期:2004-06-07 修回日期:1900-01-01 出版日期:2005-05-10 发布日期:2005-05-10
  • 通讯作者: 田见晖

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

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  1. 中国农业大学动物科技学院
  • Received:2004-06-07 Revised:1900-01-01 Online:2005-05-10 Published:2005-05-10

摘要: 探索了离子霉素结合细胞松驰素B(cytochalasin B, 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、30 mol·L-1的离子霉素处理40 min或电激活(1.3 kV·cm-1,80 s,1次脉冲),结果表明,20 mol·L-1组的激活率(69.93±5.80)%显著高于15 mol·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20 mol·L-1离子霉素分别激活10、20、30、40、50 min,再用2 mmol·L-1 6-DMAP处理6 h,其中,40 min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(20 mol·L-1、40 min)激活后,分别用7.5 g·ml-1 CB、10 g·ml-1 CHX、2 mmol·L-1 6-DMAP、7.5 g·ml-1 CB +10 g·ml-1 CHX和7.5 g·ml-1 CB +2 mmol·L-1 6-DMAP处理6 h,2 mmol·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7.5 g·ml-1 CB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(20 mol·L-1、40 min)激活后,用2 mmol·L-1 6-DMAP分别处理3.5、5.5、7.5 h,5.5 h组的卵裂率和囊胚率[(66.59±14.36)%和(25.40±10.16)%]高于另外两组,但差异不显著(P >0.05)。结果表明,离子霉素(20 mol·L-1、40 min)+6-DMAP(2 mmol·L-1 、5.5 h)为最佳激活方案。离子霉素激活卵母细胞后,CB与CHX、6-DMAP的互作对卵子的发育有抑制作用。

关键词: 猪, 卵母细胞, 体外成熟, 孤雌激活, 离子霉素

Abstract: 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 mol·L-1. Activation rates of 20, 25 mol·L-1 and 30 mol·L-1 treatments were higher (P<0.05)than that of 15 mol·L-1 treatment. In experiment 2, in vitro matured oocytes were activated with 20 mol·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 g·ml-1 CB,10 g·ml-1 CHX, 2 mmol·L-1 6-DMAP, 7.5 g·ml-1 CB +10 g·ml-1 CHX or 7.5 g·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 7.5 g·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 mol·L-1, 40 min) followed by 6-DMAP (2 mmol·L-1 , 5.5 h).

Key words: Porcine, Oocyte, in vitro maturation, Parthenogenetic activation, Ionomycin