Huan Yan Jun, Wu Zhan Feng, Zhang Ji Guang, Zhu Jiang, Xie Bing Teng, Wang Jian Yu, Li Jing Yu, Xue Bing Hua, Kong Qing Ran, Liu Zhong Hua
College of Life Science, Northeast Agricultural University, Harbin 150030, China.
J Reprod Dev. 2016;62(1):71-7. doi: 10.1262/jrd.2015-048. Epub 2015 Nov 3.
Nuclear reprogramming induced by somatic cell nuclear transfer is an inefficient process, and donor cell DNA methylation status is thought to be a major factor affecting cloning efficiency. Here, the role of donor cell DNA methylation status regulated by 5-aza-2'-deoxycytidine (5-aza-dC) or 5-methyl-2'-deoxycytidine-5'-triphosphate (5-methyl-dCTP) in the early development of porcine cloned embryos was investigated. Our results showed that 5-aza-dC or 5-methyl-dCTP significantly reduced or increased the global methylation levels and altered the methylation and expression levels of key genes in donor cells. However, the development of cloned embryos derived from these cells was reduced. Furthermore, disrupted pseudo-pronucleus formation and transcripts of early embryo development-related genes were observed in cloned embryos derived from these cells. In conclusion, our results demonstrated that alteration of the DNA methylation status of donor cells by 5-aza-dC or 5-methyl-dCTP disrupted nuclear reprogramming and impaired the developmental competence of porcine cloned embryos.
体细胞克隆核移植诱导的核重编程是一个低效过程，供体细胞的DNA甲基化状态被认为是影响克隆效率的主要因素。在此，研究了由5-氮杂-2'-脱氧胞苷（5-aza-dC）或5-甲基-2'-脱氧胞苷-5'-三磷酸（5-甲基-dCTP）调控的供体细胞DNA甲基化状态在猪克隆胚胎早期发育中的作用。我们的结果表明，5-aza-dC或5-甲基-dCTP显著降低或增加了整体甲基化水平，并改变了供体细胞中关键基因的甲基化和表达水平。然而，源自这些细胞的克隆胚胎的发育受到了抑制。此外，在源自这些细胞的克隆胚胎中观察到假原核形成紊乱以及早期胚胎发育相关基因的转录本。总之，我们的结果表明，5-aza-dC或5-甲基-dCTP改变供体细胞的DNA甲基化状态会破坏核重编程并损害猪克隆胚胎的发育能力。
