Huang Xia, Song Lili, Zhan Zhiyan, Gu Haihui, Feng Haizhong, Li Yanxin
1 Key Laboratory of Pediatric Hematology and Oncology Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, School of Medicine, Shanghai Jiao Tong University , Shanghai, China .
2 Department of Transfusion Medicine, Shanghai Changhai Hospital , Shanghai, China .
Cell Reprogram. 2017 Dec;19(6):344-353. doi: 10.1089/cell.2017.0021. Epub 2017 Nov 14.
Successful development of interspecies somatic cell nuclear transfer (iSCNT) embryos depends on compatibilities between ooplasmic and nuclear components. However, the mechanisms by which the compatibilities are regulated are still unknown. In this study, using mouse Oct4-green fluorescent protein (GFP) cells as donors and rabbit oocytes as recipients, we show that Oct4 and other pluripotency related genes were reactivated in some of mouse-rabbit iSCNT embryos, which could also activate Oct4 promoter-driven GFP reporter gene expression. Series nuclear transfer improved the efficiency of Oct4 reactivation. DNA demethylation of Oct4 promoter was detected in GFP positive iSCNT blastocysts, whereas GFP negative iSCNT embryos showed a low efficiency. Our results demonstrate that Oct4-GFP can well label the embryos with epigenetic remodeling and reactivation of pluripotent gene expression. Abundant rabbit mitochondria specific DNAs were identified in reconstructed mouse-rabbit embryos throughout preimplantation stages. Our data demonstrate that epigenetic remodeling and the complete mitochondrial match are not necessary for successful iSCNT embryo development before implantation.
种间体细胞核移植（iSCNT）胚胎的成功发育取决于卵质和核成分之间的兼容性。然而，调节这些兼容性的机制仍然未知。在本研究中，我们以小鼠八聚体结合转录因子4（Oct4）-绿色荧光蛋白（GFP）细胞作为供体，兔卵母细胞作为受体，结果显示在一些小鼠-兔iSCNT胚胎中Oct4和其他多能性相关基因被重新激活，这些胚胎还能激活Oct4启动子驱动的GFP报告基因表达。系列核移植提高了Oct4重新激活的效率。在GFP阳性的iSCNT囊胚中检测到Oct4启动子的DNA去甲基化，而GFP阴性的iSCNT胚胎效率较低。我们的结果表明，Oct4-GFP可以很好地标记具有表观遗传重塑和多能基因表达重新激活的胚胎。在整个植入前阶段的重构小鼠-兔胚胎中鉴定出大量兔线粒体特异性DNA。我们的数据表明，表观遗传重塑和完全的线粒体匹配对于植入前iSCNT胚胎的成功发育并非必需。
