SMYD3 对克隆猪早期胚胎发育的影响研究  

Study on the Effect of SMYD3 on theEarly Embryonic Development of Cloned Pigs

作　　者：恽雪丹 张艳[1] 黄时海[1] 曹丽华[1] 杨婷[1] 石德顺[1] 李湘萍[1] YUN Xue-dan;ZHANG Yan;HUANG Shi-hai;CAO Li-hua;YANG Ting;SHI De-shun;LI Xiang-ping(State Key Laboratory of Agro-biological Resources Protection and Utilization in Subtropical Region,Guangxi University,Nanning 530004,China)

机构地区：[1]亚热带农业生物资源保护与利用国家重点实验室广西大学,南宁530004

出　　处：《西南大学学报（自然科学版）》2021年第7期61-67,共7页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金项目(32060754);广西自然科学基金项目(2020GXNSFAA238039,2019GXNSFDA185005,2019GXNSFDA185005);广西创新驱动发展专项(AA17204051).

摘　　要：目的:检测并分析SMYD3基因在猪卵母细胞体外成熟、孤雌激胚胎与体细胞核移植早期胚胎中的表达模式,研究SMYD3基因表达对猪核移植早期胚胎发育及多能性基因表达的影响.方法:通过在猪体细胞核移植胚胎中过表达和干扰SMYD3基因,观察卵裂率、四细胞率和囊胚率,并采用RT-qPCR检测不同处理组各时期SMYD3的表达情况以及多能性基因Nanog和Oct4的表达量.结果:qRT-PCR结果显示,SMYD3基因在猪卵母细胞体外成熟过程中表达较低;在孤雌激活和核移植早期胚胎发育过程中均呈现先升高后降低的表达规律,其中在4细胞和8细胞期胚胎中表达较高.过表达SMYD3基因显著提高猪核移植胚胎的囊胚率(14.63%vs 9.68%,p<0.05),下调SMYD3基因表达显著降低囊胚率(6.74%vs 9.68%,p<0.05).上调SMYD3组猪核移植囊胚Nanog和Oct4基因表达水平显著提高(p<0.05);下调组囊胚中Nanog基因检测不出,Oct4基因表达无显著差异.结论:上调SMYD3基因表达可提高猪体细胞克隆的囊胚发育率,提高核移植胚胎体外发育能力.Objective:To detect the expression pattern of SMYD3 gene in porcine oocytes in vitro maturation,parthenogenetic activation of embryos and somatic cell nuclear transfer early embryos,and to study the effect of regulating SMYD3 gene expression on porcine nuclear transfer early embryo development and pluripotency gene expression.Methods:Through overexpression and interference of SMYD3 gene in pig somatic cell nuclear transfer embryos,the lysis rate,four-cell rate and blastocyst rate were observed,and RT-qPCR was used to detect the expression of SMYD3 and pluripotency genes in different treatment groups.Results:The results of qRT-PCR showed that the expression of the SMYD3 was comparatively low during the in vitro maturation of pig oocytes.In the early embryonic development process of parthenogenetic activation and nuclear transfer,the expression of the SMYD3 gene first increased and then decreased,and the expression of the SMYD3 gene was higher in embryos at the 4-cell and 8-cell stages.Overexpression of the SMYD3 gene significantly increased the blastocyst rate of the pig nuclear transfer embryos(14.63%vs 9.68%,p<0.05),while down-regulation of SMYD3 gene expression significantly reduced the blastocyst rate(6.74%vs 9.68%,p<0.05).The expression levels of Nanog and Oct4 genes in pig nuclear transfer blastocysts in the up-regulated SMYD3 group were significantly increased(p<0.05);Nanog was not detected in down-regulated blastocyst,and the expression of Oct4 showed no significant difference.Conclusion:Up-regulation of SMYD3 gene expression can increase the blastocyst rate and improve the in vitro development of pig nuclear transfer embryos.

关 键 词：SMYD3 SHRNA 体细胞核移植 猪卵母细胞体外成熟 

分 类 号：S828[农业科学—畜牧学]