密闭培养条件下小鼠早期胚胎Igf2/H19印迹调控区甲基化状态分析  

Methylation of Igf2/H19 Imprinting Control Region in Mouse Early Embryos Cultured in Sealed Conditions

作　　者：陈海燕[1] 马保华[1] 刘洁[1] 赵晓娥[1] 曹宇静[2] 雷晓华[2] 

机构地区：[1]西北农林科技大学动物医学院农业部动物生殖生理和胚胎工程重点开放实验室,陕西杨凌712100 [2]中国科学院动物研究所生殖生物学国家重点实验室,北京100101

出　　处：《中国生物化学与分子生物学报》2010年第7期672-677,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家空间环境利用项目(NoSJ-10B3)~~

摘　　要：胚胎密闭培养是空间胚胎发育研究的基本条件.本文主要研究密闭培养条件对小鼠早期胚胎发育过程中印迹基因Igf2/H19的印迹调控区(ICR)甲基化水平的影响.应用亚硫酸氢盐测序法(BSP)分析小鼠2-细胞胚胎在密闭条件下分别培养24h、48h和72h后,相对应的发育阶段胚胎Igf2/H19的ICR甲基化水平,以常规体外培养和体内发育的各阶段胚胎为对照.结果显示,密闭培养条件下,小鼠8-细胞胚胎、桑葚胚和囊胚的Igf2/H19的ICR甲基化水平都低于常规体外培养的结果,且更明显低于体内发育的结果;同时发现,小鼠8-细胞胚胎密闭培养时,Igf2/H19的ICR甲基化水平最低.由此可见,密闭培养会引起小鼠植入前各发育阶段胚胎Igf2/H19的ICR甲基化水平降低,并证明Igf2/H19的ICR甲基化水平可以作为监测哺乳动物早期胚胎发育状况的分子指标.The sealed-culture of embryo is the basic condition for the embryo development studies in space.The study was mainly the impaction of the methylation levels of Igf2/H19 imprinting control region（ICR） in the development process of the mouse early embryos under sealed-culture,with general-culture in vitro and in vivo development as control.Using the bisulfite sequencing PCR（BSP） to analyze the different methylation levels of Igf2/H19 ICR in every stage of embryo development,2-cell mouse embryos were cultured for 24,48,and 72 hours in sealed-culture.The result showed that the methylation levels of Igf2/ H19 ICR in mouse 8-cell embryos,morula,blastocyst cultured in sealed-culture all were lower than those of general cultured in vitro,and significantly lower than those of development in vivo while the methylation level of mouse 8-cell embryos cultured in sealed-culture was the lowest.We concluded that sealed-culture made the methylation levels of Igf2/H19 imprinting control region of mouse preimplantation embryos decreased,and the study also had confirmed that the methylation level of Igf2/H19 imprinting control region could be as a molecular target to detect the development status of the mammal early embryos.

关 键 词：密闭培养 胚胎 甲基化 印迹基因Igf2/H19 小鼠 

分 类 号：Q13[生物学—普通生物学] Q78