热应激对牛卵母细胞克隆胚体外发育的影响  

Effect of heat shock on nuclear transferred bovine oocytes development in vitro

作　　者：张弛[1] 曹俊伟[1] 张涌[1] 

机构地区：[1]西北农林科技大学生物工程研究所,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2007年第12期6-10,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"高技术研究与发展计划项目(2001AA21380)

摘　　要：为了了解牛卵母细胞经短时间(30 min)热应激和采用不同供体细胞对核移植后重构胚发育的影响,对成熟前牛卵母细胞进行了不同温度(38.5(CK),39.5,40.5,41.5和42.5℃)热应激处理,以牛胎儿成纤维细胞为供体细胞,比较了重构胚的卵裂率和囊胚率;将成熟前牛卵母细胞于41.5℃热应激30 min,以未进行热应激处理的牛卵母细胞为对照,比较了3种不同供体细胞(牛胎儿成纤维细胞、牛卵巢上皮细胞、牛颗粒细胞)核移植后的重构胚囊胚细胞数。结果表明,成熟前牛卵母细胞经41.5℃热应激处理30 min,其卵裂率(72.5%)和囊胚率(18.3%)与对照组(57.6%和13.6%)相比均有显著提高(P<0.05);当温度升高至42.5℃后,虽然卵裂率有所提高,但囊胚率与对照组相比显著降低(P<0.05);其他温度处理的卵裂率和囊胚率与对照无显著差异(P>0.05)。牛卵母细胞经41.5℃热应激处理30 min,3种供体细胞形成的重构胚囊胚细胞数差异不显著(P>0.05),而且与其各自对照的囊胚细胞数差异也不显著(P>0.05)。说明41.5℃可以作为牛卵母细胞热应激处理的最佳温度,成熟前牛卵母细胞经41.5℃热应激处理30 min,能有效提高核移植效率。To understand the effect of short-time （30 min） heat shock with bovine recipient oocytes and various donor cell on development of early nuclear transfer embryos, with the bovine fetal fibroblast cell as donor cell,the cleavage rate and blastocyst rate of cloned embryos were compared by heat shocked bovine oocytes before maturation at different temperature（ 38.5,39.5,40.5,41.5 and 42.5 ℃）;the number of blastocyst cell was compared after reconstructing of three various donor cells（the fibroblast cell,the epi- thelial cell and the granulose cell） with the bovine oocytes which did not have a heat-shock treatment at 41.5 ℃ in 30 min. Experimental results showed that a heat-shock treatment at 41.5℃ could improve the cleavage rate（72.5%） and blastocyst rate（18.3%） of cloned embryo at the early stage in comparison with the control group （57.6 %and 13.6%） （P〈0.05） ;after the heat-shock treatment at 42.5 ℃ ,the cleavage rate increased,however, the blastocyst rate decreased significantly in comparison with the control group （P〈0.05） ;the cleavage and the blastocyst rate did not have change in comparison with the control group （P〉0.05） under other temperate heat-shock. Using the bovine oocytes under a heat-shock treatment at41.5 ℃,the number of blastocyst cell formed by three various donor cells （P〉0.05） did not have signifi cant difference. As a result,it was concluded that 41.5 ℃ was the best temperature for heat-shock treat- ment on recipient oocytes,and cloned embryo at the early stage under a heat-shock treatment at 41.5 ℃ for 30 min could lead to high bovine nuclear transfer efficiency.

关 键 词：牛 卵母细胞 热应激 细胞克隆 核移植 

分 类 号：S823.36[农业科学—畜牧学]