›› 2009, Vol. 36 ›› Issue (1): 73-76.

• 遗传繁育 • 上一篇    下一篇

不同化学激活剂对牛卵丘细胞重构胚发育的影响

王滇,张春燕,刘松柏,杨利国   

  1. 华中农业大学动物遗传育种与繁殖教育部重点实验室,武汉 430070
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-01-20 发布日期:2009-01-20

The Effect of Different Activation Methods on Bovine Reconstructed Embryos Produced Using Cumulus Cells

WANG Dian,ZHANG Chunyan, LIU Songbai,YANG Liguo   

  1. Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan 430070, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-01-20 Published:2009-01-20

摘要: 本研究主要研究了Ⅰ:Ionomycin+6甲氨基嘌呤(6DMAP)、Ⅱ:Ionomycin+放线菌酮(CHX)、Ⅲ:体积分数7%乙醇+6DMAP、Ⅳ:体积分数7%乙醇+CHX 4种不同化学激活方法对牛胞质内卵丘细胞全细胞注射法所获得重构胚的激活及前期发育的影响。Ionomycin和7%乙醇的处理时间为5 min, 6DMAP的处理时间为4 h,CHX的处理时间为5 h。结果表明,重构胚与颗粒细胞单层细胞共培养2 d后,各组卵裂率分别为52.6%、52.8%、53.8%和54.2%,差异不显著(P>0.05);培养8 d后,Ⅰ组激活的囊胚发育率(20.0%)极显著高于其他组(P<0.01),其他各组之间差异不显著,分别为8.5%、10.2%和6.1%;培养10 d后,Ⅰ组激活的囊胚孵化率(10.7%)极显著高于其他组(P<0.01),其他各组之间差异不显著,分别为2.3%、3.0%和1.8%。结果提示,4种激活方法均可以有效的激活重构胚,但Ionomycin+6DMAP法激活重构胚的囊胚率和囊胚孵化率均极显著高于其它3组,说明Ionomycin+6DMAP激活法有利于牛重构胚的发育,可以获得较理想的囊胚率,是胞质内全细胞注射法克隆的理想激活方法。

关键词: 牛; 卵母细胞; 胞质内直接注射; 重构胚胎; 化学激活

Abstract: The objective of this article was to investigate the effect of 4 different chemical activation reagents on the rate of development of bovine reconstructed embryos produced by intracytoplasmic direct injection. The reconstucted embryos were randomly divided into 4 groups to be activated:
Ionomycin+6DMAP; Ionomycin+Cycloheximide (CHX);7% ethanol+6DMAP;7% ethanol+CHX. The reconstructed embryos were activated for 5 min in Ionomycin and 7% ethanol. 6DMAP,4 h; CHX, 5 h. The reconstructed embryos were cocultured with cumulus cell monolayer. The rate of cleavage of the 4 groups were 52.6%,52.8%,53.8% and 54.2%, respectively. There was no significant difference among the 4 groups in terms of the rate of cleavage. The rate of blastocyst at day 8 were 20.0%,8.5%,10.2% and 6.1% for groupⅠ,Ⅱ,Ⅲ and Ⅳ, respectively. The rate of blastocyst of group Ⅰ was significantly higher than that of three other groups. The rate of hatching blastocyst of group Ⅰ showed very significant difference with that of group Ⅱ,Ⅲ and Ⅳ (10.7%,2.3%,3.0% and 1.8%, respectively, P<0.01). The results showed that all of the 4 kinds of activation reagents could successfully activated the reconstructed bovine embryos. The group Ⅰ gained the highest rate of blastocyst and hatching blastocyst, which demonstrated that Ionomycin+6DMAP benefited the development of bovine reconstructed embryos and could obtain an ideal rate of blastocyst. Ionomycin+6DMAP were ideal activation reagents for bovine reconstructed embryos produced by intracytoplasmic direct injection.

Key words: bovine; oocyte; intracytoplasmic direct injection; reconstructed embryo; chemical activation

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