雷帕霉素对小鼠体外受精胚胎早期发育的影响  

Effects of Rapamycin on Early Development of Mouse InVitroFertilized Embryos

作　　者：周亮[1] 张喜凤[1] 周娜汝[1,2] 吴蓉花[1,2] 李运生[1,2] 张宇[1,2] 张运海[1,2] 

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]安徽地方畜禽遗传资源保护与生物育种省级实验室,安徽合肥230036

出　　处：《动物医学进展》2013年第12期101-105,共5页Progress In Veterinary Medicine

基　　金：大学生科技创新基金项目(2011188)

摘　　要：以小鼠为模型,探索了哺乳动物雷帕霉素靶蛋白(mTOR)特异性抑制剂雷帕霉素(RAPA)在小鼠体外受精胚胎早期发育过程中生理学作用。分别使用添加不同剂量(对照组0ng/mL;试验组0.1,1,10ng/mL和100ng/mL)RAPA的化学限定培养液连续培养小鼠体外受精原核期与2细胞期胚胎,检测其体外发育率(卵裂率、囊胚率)及囊胚质量(囊胚内细胞团细胞数、滋养层细胞数、总细胞数)。结果显示,在向原核期开始体外培养的受精卵培养液中添加不同浓度的RAPA,体外培养至24h时,各处理组的卵裂率与对照组差异不显著(P>0.05),受精卵卵裂未受影响;而在原核期及2细胞期开始培养的胚胎培养液中加入不同浓度的RAPA,培养90h^96h后,发现100ng/mL RAPA组的囊胚率显著低于对照组(P<0.05)和其他处理组(P<0.05)。囊胚细胞计数结果表明,100ng/mLRAPA处理组囊胚内细胞团细胞数、滋养层细胞数及总细胞数均显著减少(P<0.05)。研究结果表明,雷帕霉素添加量达到100ng/mL时开始出现对小鼠早期胚胎体外发育率和发育质量的显著抑制作用,同时表明在小鼠胚胎早期发育过程中,mTOR信号通路对维持小鼠胚胎囊胚的正常形成是必不可少的。This study was to explore the physiological function of rapamycin（RAPA）,the specific inhibitor of mammalian target of rapamycin（mTOR）,during early development of mouse embryos derived fromin vitro fertilization（IVF）.Chemically defined medium with different concentrations of RAPA（the control group：0ng/mL;the treatment groups：0.1,1,10 and 100ng/mL）was used for embryo culture,and the in vitro cleavage rate,formation rate and quality of blastocysts（the number of inner cell mass（ICM）cell,trophectoderm（TE）cell and total cell）of the embryos were examined.Results were as follows.After 24 hours of embryos cultured,the cleavage rate was found no significant difference when RAPA added from the pronuclear stage embryos,but after 90-96hours of embryos cultured,the blastocyst rate of 100ng/mL group was significantly lower than that of the other groups（including the control group）（P〈0.05）when RAPA added both from the pronuclear stage embryos and 2-cell stage embryos.The result of blastocyst cell counting showed,the 100ng/mL group was found obviously decreased compared to the control group（P〈0.05）on the number of ICM cell,TE cell and total cells,which indicated that the quality of blastocyst development was affected.Our research suggests the early development of in vitro mouse embryo was restrained by RAPA at the beginning concentration of 100ng/mL,and the mTOR signal pathway within the maintenance of mouse blastocyst formation is essential.

关 键 词：雷帕霉素 MTOR信号通路 囊胚形成 体外受精胚胎 小鼠 

分 类 号：S852.2[农业科学—基础兽医学]