6-DMAP对延边黄牛末期卵母细胞核移植的影响

Effect of 6-DMAP on Nuclear Transplantation of Yanbian Cattle Telophase Oocytes

为探讨6-二甲基氨基嘌呤(6-DMAP)在延边黄牛末期去核的体细胞克隆中的作用,本试验主要研究了单独使用离子霉素处理与离子霉素联合添加6-DMAP处理对体外成熟的老化延边黄牛卵母细胞的激活率的不同影响;以及不同时期添加6-DMAP对重构胚后期发育能力的影响。试验结果显示,体外成熟的老化卵母细胞,使用离子霉素单独处理后91%被激活,而在离子霉素联合添加6-DMAP处理后却没有细胞被激活,也就是没有第二极体的排出。另外,融合后使用6-DMAP处理,所得重构胚的囊胚发育率最低,而激活后的卵母细胞立即用6-DMAP处理,所得重构胚的发育能力与无6-DMAP处理的相近。综上所述,离子霉素联合添加6-DMAP可抑制老化的延边黄牛卵母细胞的激活,阻止第二极体的排出,但对重构胚的后期发育没有影响。而融合使用添加6-DMAP的培养液,抑制了重构胚的后期发育。

In order to researching the effect of 6-dimethylaminopurine （6-DMAP） on the nuclear transplantation of Yanbian cattle telophase oocytes, we studied the different effect on the activation rate of Yanbian cattle telophase oocytes which were treated with Ionomycin alone and followed with 6-DMAP, and the effect of different period treatment with 6-DMAP on the development of their reconstructed embryos.The result showed that 91% in vitro matured aged oocytes which were treated with Ionomycin alone could be acted, but none would be acted which were treated with Ionomycin and followed with 6-DMAP, and the polar body extrusion was inhibited. Additionally, the blastula rate of reconstructed embryos which were treated with 6-DM-AP treatment followed fusion was the lowest, however the developmental capacity of reconstructed embryos of the oocytes which immediately treated with 6-DMAP followed the activation was close to those which had no treatment with 6-DMAP. These results demon-strated that the treatment of Ionomycin combining with 6-DMAP can inhibit the activation of Yanbian cattle telophase oocytes, and the polar body extrusion was inhibited, but had no effect on the developmental capacity of reconstructed embryos. But adopting the culture fluid which added 6-DMAP can inhibit the developmental capacity of reconstructed embryos.