p66Shc对绵羊早期胚胎发育的影响  

Effects of p66Shc on the Early Embryo Development of Sheep

作　　者：赵勇超 张通 王兆琛 刘春洁 张文广[1] 张家新[1] ZHAO Yong-chao;ZHANG Tong;WANG Zhao-chen(College of Animal Science,Inner Mongolia Agricultural University/Inner Mongolia Key Laboratory of Animal Genetics,Breeding and Reproduction,Hohhot,Inner Mongolia 010018;College of Medicine,Shanxi Datong University,Datong,Shanxi 037009)

机构地区：[1]内蒙古农业大学动物科学学院/内蒙古自治区动物遗传育种与繁殖重点实验室,内蒙古呼和浩特010018 [2]山西大同大学医学院,山西大同037009

出　　处：《安徽农业科学》2021年第20期108-112,133,共6页Journal of Anhui Agricultural Sciences

基　　金：内蒙古自治区科技重大专项(2020ZD0003)。

摘　　要：[目的]探讨p66Shc对绵羊早期胚胎发育的影响。[方法]以受精后28 h为节点,将早期胚胎分为早卵裂胚胎和晚卵裂胚胎,观察不同时期卵裂胚胎的进一步发育情况,同时通过实时荧光定量PCR检测不同来源的4-8细胞阶段胚胎中p66Shc基因的表达水平;利用siRNA分子干扰技术将特异性siRNA分子通过显微注射到绵羊合子细胞质中,以敲减早期胚胎p66Shc基因;通过免疫荧光技术检测p66Shc基因对绵羊早期胚胎发育过程中活性氧(ROS)和氧化应激标记物(8-OHdG)表达的影响。[结果]早卵裂胚胎的囊胚发育率显著高于晚卵裂胚胎(P<0.05);晚卵裂胚胎中p66Shc mRNA表达水平显著高于早卵裂胚胎(P<0.05);将绵羊早期胚胎中的p66Shc基因敲减后,与对照组相比,其胚胎内p66Shc mRNA水平、p66Shc蛋白水平显著降低,ROS和8-OHdG水平显著低于对照组(P<0.05);基因敲减后桑椹胚发育率显著升高(P<0.05)。[结论]低水平的p66shc可以提高绵羊早期胚胎对氧化应激的抵抗力,并进一步提高其发育能力。[Objective]To explore the effects of p66Shc on early embryo development of sheep.[Method] The early embryos were divided into early cleavage(EC) embryo and late cleavage(LC) embryo at 28 h after fertilization(28 hpi).The further development of cleavage embryos at different stages were observed.At the same time, the expression level of p66Shc gene in 4-8 cell stage early embryos from different sources was detected by real-time quantitative PCR.Using siRNA molecular interference technology, specific siRNA molecules were microinjected into the cytoplasm of sheep zygotes, so as to knock down p66Shc gene of early embryos.The effects of p66Shc gene on reactive oxygen species(ROS) and oxidative stress marker(8-OHDG) during the early embryo development process of sheep were investigated by using immunofluorescence technique.[Result]The blastocyst development rate of EC embryos was significantly higher than that of LC embryos(P<0.05).The expression level of p66Shc mRNA in LC embryos was significantly higher than that in EC embryos(P<0.05).After knocking down p66Shc interference, p66Shc mRNA and protein levels in early sheep embryos were significantly decreased than those in control group(P<0.05),the expression levels of ROS and 8-OHdG in early sheep embryos were lower significantly than those in control group(P<0.05).Moreover, morula development rate was significantly increased(P<0.05)after knocking down p66Shc gene.[Conclusion]Low level of p66Shc could improve the resistance of early sheep embryos to oxidative stress and further improve their developmental ability.

关 键 词：绵羊 P66SHC 早期胚胎 卵裂 氧化应激 

分 类 号：S826[农业科学—畜牧学]