3种来源猪胚胎激活后6h内基因组DNA甲基化变化模式  

DNA methylation pattern of three types of pig embryos within 6 hours after activation

作　　者：任子利[1,2,3] 赵彦玲[1,2,3] 杨小淦[1,2] 陆阳清[1,2] 卢晟盛[1,2] 卢克焕[1,2] 

机构地区：[1]广西大学广西亚热带生物资源保护利用重点实验室,广西南宁530004 [2]广西大学动物科技学院,广西南宁530004 [3]西藏农牧学院动物科技学院,西藏林芝860000

出　　处：《中国兽医学报》2011年第3期420-424,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30660127;30860039);广西科学研究与技术开发计划资助项目(0815011-6-3)

摘　　要：主要探讨猪手工克隆(HMC)胚胎和体外受精(IVF)胚胎及孤雌激活(PA)早期胚胎发育过程中6 h内的单细胞核全基因组DNA甲基化模式。运用抗5-甲基胞嘧啶抗体免疫荧光和激光共聚焦显微镜检测相对荧光强度的方法,检测猪手工克隆胚在融合后6 h内全基因组DNA甲基化模式,并与同时期猪体外受精胚及孤雌激活胚胎进行比较,从而探讨这3种胚胎变化模式的异同。结果显示:3种不同来源的胚胎在培养后不同时间点比较时,2、4、6 h均呈强甲基化状态,且均呈下降趋势;在同时间点比较时,2 h和6 h这两个时间点检测到3种胚胎的相对荧光强度均差异不显著(P>0.05),而在4 h时,HMC胚胎显著高于PA和IVF胚(P<0.05)。结论:猪手工克隆胚在融合后6h内虽然也发生类似IVF胚的全基因组DNA去甲基化,但其去甲基化不充分,甲基化程度偏高,这可能是导致核移植胚胎核重编程不充分、发育能力低的一个原因。The objective of this study was to determine the individual nuclei DNA methylation pattern of porcine HMC（hand-made cloning）,IVF（in vitro fertilization） and PA（parthenogenetic activation） embryos during the early embryogenesis.The levels of DNA methylation were semi-quantitative by confocal microscopy through measuring the fluorescent intensity of the developmental stages embryos after using double immunostaining by the anti-5-methyl cytosine.The DNA methylation of HMC embryos were analysed at 2,4,6 h after embryos fusion and activation,compared with IVF and PA embryos.High methylation level of DNA in pig embryos from HMC,IVF and PA was observed at 2 h,but decreased when the culture time was prolonged.There was no significant difference in the methylation level among HMCIVF and PA embryos at 2 h and 6h（P0.05）,but significantly higher in HMC embryo than that of IVF and PA embryos while cultured for 4 h（P0.05）.High DNA methylation at 4 h indicated the insufficient reprogramming of porcine HMC embryos,which could result in a low competence of reconstructed embryo development.

关 键 词：猪 卵母细胞 手工核移植 DNA甲基化 

分 类 号：S827[农业科学—畜牧学] S814.8[农业科学—畜牧兽医]