H3K4me3对猪卵母细胞体外成熟和早期胚胎发育的影响  

Effects of H3K4me3 on in vitro Maturation ofPorcine Oocytes and Early Embryo Development

作　　者：曾毅仁 张彩玉 侯晗琦 恽雪丹 李湘萍[1] ZENG Yi-ren;ZHANG Cai-yu;HOU Han-qi;YUN Xue-dan;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45-51,共7页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金项目(32060754,31560632);广西自然科学基金项目(2020GXNSFAA238039).

摘　　要：哺乳动物卵母细胞的成熟与组蛋白甲基化修饰密切相关.CPI-455是一种组蛋白去甲基化酶抑制剂,可特异性抑制组蛋白去甲基转移酶KDM5A/B的活性,提高H3K4me3(组蛋白H3第4位赖氨酸三甲基化)的表达水平.本实验在猪卵母细胞体外成熟过程中添加CPI-455,探究H3K4me3组蛋白甲基化对猪卵母细胞体外成熟和早期胚胎发育的影响.首先,用不同浓度CPI-455在不同时间处理猪卵母细胞,发现在0~22 h添加5μmol CPI-455的处理方法效果最佳,卵母细胞成熟率与囊胚形成率均显著高于未处理组(p<0.05).0~22 h添加5μmol CPI-455处理卵母细胞显著提高了囊胚中H3K4me3的表达(p<0.05),降低了卵母细胞中KDM5A和KDM5B的表达(p<0.05),并且显著提高了囊胚中Nanog,Oct4,CDX2等多能性基因的表达(p<0.05).CPI-455处理可以通过降低卵母细胞中相关基因的表达,上调囊胚中多能性基因的表达,调控H3K4me3的甲基化水平,从而促进猪卵母细胞体外成熟和早期胚胎发育.The maturation of oocytes is closely related to histone methylation modification in mammalian species.CPI-455 is a histone demethylase inhibitor,which can specifically inhibit the activity of histone demethyltransferase KDM5A/B and improve the expression level of H3K4me3.In this study,CPI-455 was added into the in vitro maturation medium of porcine to investigate the effect of H3K4me3 histone methylation on in vitro maturation and early embryonic development of porcine oocytes.First of all,oocytes were treated with CPI-455 based on different concentrations in different times.It was found that 5μmol CPI-455 added in 0-22 h in vitro maturation significantly increased oocyte maturation rate and subsequent embryo formation rate(p<0.05).Therefore,this treatment method was adopted in subsequent experiments.The results showed that the expression level of H3K4me3,KDM5A and KDM5B in oocytes were significantly increased(p<0.05),the expressions of pluripotent genes,such as Nanog,Oct4 and CDX2,in blastocysts were significantly increased in oocytes treated with CPI-455(p<0.05).In conclusion,CPI-455 treatment can reduce the expression of related genes in porcine oocytes,up-regulate the expression of pluripotent genes in the blastocysts,and regulate the methylation level of H3K4me3,so as to promote in vitro maturation and early embryonic development of porcine oocytes.

关 键 词：猪 卵母细胞 CPI-455 H3K4me3 体外成熟 

分 类 号：S828[农业科学—畜牧学]