毛壳素对猪核移植胚胎体外发育潜能的影响研究  

Effects of chaetocin treatment on in vitro developmental potential of nuclear transfer embryos of pig

作　　者：王婷[1] 李海艳[1] 刘晨[1] 孙乐[1] 杨婷[1] 石德顺[1] 李湘萍[1] WANG Ting;LI Hai-yan;LIU Chen;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6期792-797,共6页Chinese Veterinary Science

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

摘　　要：为了解H3K9三甲基化表达水平对德保猪核移植胚胎发育潜能的影响,本试验使用毛壳素处理德保猪胎儿成纤维细胞,利用体细胞核移植技术观察猪核移植胚胎的体外发育和相关基因mRNA表达情况。结果显示,不同浓度毛壳素处理(20、30、50 nmol/L)德保猪成纤维细胞24 h时,20 nmol/L处理组的细胞生长曲线与未处理组相似,呈“S”型。实时定量PCR检测结果显示,20 nmol/L和30 nmol/L毛壳素处理均会显著降低SUV39H1/2和G9a在成纤维细胞中的表达(P<0.05)。核移植胚胎体外发育结果显示,与未处理组相比,20 nmol/L处理可显著提高核移植胚胎的4-细胞发育率和囊胚发育率(P<0.05);30 nmol/L处理的卵裂率、4-细胞发育率和囊胚发育率显著降低(P<0.05)。实时定量PCR检测结果显示,与未处理组相比,20 nmol/L处理可显著降低SUV39H1/2和G9a在2-细胞、4-细胞和8-细胞期核移植胚胎中的表达,显著提高2-细胞、4-细胞和核移植胚胎中eIF3A和TFIIA、囊胚中Nanog和Oct-4基因的表达(P<0.05)。研究结果表明:适宜浓度的毛壳素处理可降低德保猪核移植胚胎中H3K9me3的表达,提高核移植早期胚胎的发育潜能。In order to understand the effects of H3K9me3 expression level on the developmental potential of Debao pig nuclear transfer embryos,this study treated Debao pig fetal fibroblasts with chaetocin,and observed in vitro development of pig nuclear transfer embryos and mRNA expression of related genes by using somatic cell nuclear transfer technology.The results showed that the cell growth curve of Debao pig fibroblasts treated with different concentrations of chaetocin(20,30,50 nmol/L)for 24 h in 20 nmol/L group was similar to that of the untreated group,showing an“S”type.The results of real-time quantitative PCR showed that the expression of SUV39H1/2 and G9A in fibroblasts were significantly decreased by 20 nmol/L and 30 nmol/L chaetocin treatment(P<0.05).The results of in vitro observation of the development of nuclear transfer embryos showed that,compared with untreated group,treatment at 20 nmol/L significantly increased the 4-cell development rate and blastocyst development rate of nuclear transfer embryos(P<0.05).The cleavage rate,4-cell development rate and blastocyst development rate of 30 nmol/L treatment were significantly decreased(P<0.05).Real-time quantitative PCR results showed that,compared with the untreated group,20 nmol/L treatment significantly decreased the expression of SUV39H1/2 and G9a in 2-cell,4-cell and 8-cell stage nuclear transfer embryos,and significantly increased the expression of eIF3a and TFIIA in 2-cell,4-cell and nuclear transfer embryos,and Nanog and OCT-4 in blastocysts(P<0.05).The results showed that the appropriate concentration of chaetocin treatment can reduce the expression of H3K9me3 in Debao pig nuclear transfer embryos,and improve the development potential of embryos in the early stage of nuclear transfer.

关 键 词：毛壳素 SUV39H1/2 H3K9me3 胚胎发育 

分 类 号：S852.1[农业科学—基础兽医学]