过表达YY1对水牛早期胚胎发育及DNA甲基化的影响  

Effects of overexpression YY1 on buffalo early embryonic development and DNA methylation

作　　者：任雪婷 龚云[1] 黄时海[1] 孙乐[1] 杨婷[1] 石德顺[1] 李湘萍[1] REN Xue-ting;GONG Yun;HUANG Shi-hai;SUN Le;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]广西大学亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004

出　　处：《中国兽医科学》2021年第11期1447-1452,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(32060754);广西自然科学基金项目(2020GXNSFAA238039,2019GXSFDA185005);广西创新驱动发展专项(AA17204051)。

摘　　要：为了解阴阳因子1(YY1)在水牛植入前胚胎中的表达规律及其对早期胚胎发育和DNA甲基化的影响,本研究通过受精卵卵胞质注射的方法,将线性化处理的过表达YY1质粒注射到体外受精8~10 h的水牛受精卵中,分析其对水牛早期胚胎发育的影响,并在胚胎发育各个时期检测该基因的表达。同时通过qRT-PCR检测过表达YY1对DNMT1表达的影响来研究DNA甲基化水平。qRT-PCR结果表明,YY1和DNMT1在水牛植入前胚胎的表达模式呈相反的趋势,YY1的表达随着胚胎的发育逐渐增高,囊胚期达到最高,而DNMT1基因在桑葚胚前表达较高,囊胚期表达最低。与空白对照组相比,注射25μg/mL质粒对水牛囊胚发育率无显著影响(P>0.05),取得较高的表达EGFP囊胚(33.3%),显著高于5、15、50μg/mL处理组(P<0.05);处理组与空白对照组的胚胎分裂率、囊胚发育率、囊胚基因表达率差异均不显著(P>0.05)。过表达YY1胚胎在体外受精12 d后仍有囊胚产生。与空白对照组相比,处理组各时期胚胎中YY1和DNMT1表达量均有明显的增加。以上结果表明,过表达YY1对体外受精早期胚胎发育没有影响,但会延迟囊胚的形成。To understand the expression pattern of YY1 in buffalo preimplantation embryos,and its effect on DNA methylation regulation The linearized YY1 overexpression plasmid was injected into the buffalo embryos which were at the stage of 8—10 h after IVF,its effect on buffalo early embryo development and the expression of YY1 at each stage of embryonic development was analyzed.For the study of DNA methylation level,the effect of overexpression of YY1 on the expression of DNMT1 was detected by q RT-PCR.The results showed that the expression patterns of YY1 and DNMT1 in buffalo preimplantation embryos were opposite,the expression of YY1 increased gradually with the development of embryo,and reached the highest level at blastocyst stage.Reversely,the expression of DNMT1 was higher before the morula stage,lowest in blastocyst stage.Compared with the control group,the 25μg/m L treatment had no significant difference on blastocyst rate(P>0.05),obtained a significantly higher EGFP positive blastocyst rate compared with other treatments(5,15,50μg/m L,P<0.05).There was no significant difference in embryo division rate,blastocyst development rate,and blastocyst gene expression rate between the treatment group and the control group(P>0.05),but the difference in the blastocyst development rate was extremely significant(P<0.05).Overexpressing YY1 embryos still had blastocysts 12 days after in vitro fertilization.Compared with the control group,the expression levels of YY1 and DNMT1 increased significantly in the treatment group.The results indicate that overexpression of YY1 has no influence on buffalo IVF embryo development,but delay blastocyst formation.

关 键 词：水牛 YY1基因 DNA甲基化 DNMT1基因 

分 类 号：S857.3[农业科学—临床兽医学]