小鼠四倍体早期胚胎基因组甲基化模式  

DNA methylation patterns of mouse tetraploid embryos

作　　者：员新旭[1] 冯书堂[1] 潘登科[1] 窦红伟[1,2] 牟玉莲[1] 李奎[1] 

机构地区：[1]中国农业科学院北京畜牧兽医研究所,畜禽遗传资源与利用农业部重点开放实验室,北京100193 [2]甘肃农业大学动物医学院产科室,兰州730070

出　　处：《遗传》2009年第4期387-392,共6页Hereditas（Beijing)

基　　金：国家高技术研究与发展计划项目(863计划)(编号:2006AA0110Z1039);国家自然科学基金项目(编号:30871775)资助

摘　　要：利用小鼠抗5-甲基胞嘧啶(5MeC)单克隆抗体检测了体外培养小鼠四倍体早期胚胎的基因组甲基化模式。结果表明:利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程,在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样,呈现高度甲基化状态;在细胞核开始融合的时候,甲基化水平急速下降,在细胞核完全融合的时候甲基化水平达到最低点;随着胚胎继续分裂,胚胎甲基化水平逐渐增加,在桑葚胚期甲基化水平最高;但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别,这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。因此,小鼠体外培养四倍体胚胎的甲基化模式是不正常的,这可能是四倍体小鼠难以发育到妊娠足月的原因之一。这是对小鼠四倍体早期胚胎基因组甲基化模式的首次报道。In the present study, the DNA methylation patterns of in vitro-derived mouse tetraploid embryos were investigated by immunofluorescence staining with an antibody against 5-methylcytosine （5MeC）. Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastocysts followed by fusion of cytoplasm and nucleus and cleavage in vitro. During the fusion of cytoplasm, the DNA methylation levels of the fused embryos are as high as these of two-cell diploid embryos in vivo Then the embryos are rapidly demethylated when the nucleus begin to fuse, resulting in the lowest DNA methylation levels when the nucleus are fused completely. After that, the DNA methylation levels of the fused embryos are gradually increased until the morula stage. However, whereas an asymmetric distribution of DNA methylation is established in vivo-derived blastocysts with a higher methylation level in the inner cell mass （ICM） than that in the trophectoderm, we can not detect the asymmetric distribution in most in vitro-derived tetraploid blastocysts. So the DNA methylation patterns of mouse tetraploid embryos are aberrant, which may lead to subsequent developmental failure and embryo death. This is the first report on the methylation patterns of in vitro-derived mouse tetraploid embryos.

关 键 词：小鼠 四倍体胚胎 甲基化 抗5-甲基胞嘧啶抗体 

分 类 号：Q953[生物学—动物学]