Kang Yong-Kook, Yeo Seungeun, Kim Seok-Ho, Koo Deog-Bon, Park Jung Sun, Wee Gabbine, Han Jee-Soo, Oh Keon Bong, Lee Kyung-Kwang, Han Yong-Mahn
Laboratory of Development and Differentiation, Korea Research Institute of Bioscience and Biotechnology, Daejeon, South Korea.
Mol Reprod Dev. 2003 Sep;66(1):32-7. doi: 10.1002/mrd.10330.
Change of DNA methylation during preimplantation development is very dynamic, which brings this term to the most attractive experimental target for measuring the capability of cloned embryo to reprogram its somatic genome. However, one weak point is that the preimplantation stage carries little information on genomic sequences showing a site-specific re-methylation after global demethylation; these sequences, if any, may serve as an advanced subject to test how exactly the reprogramming/programming process is recapitulated in early cloned embryos. Here, we report a unique DNA methylation change occurring at bovine neuropeptide galanin gene sequence. The galanin gene sequence in early bovine embryos derived by in vitro fertilization (IVF) maintained a undermethylated status till the morula stage. By the blastocyst, certain CpG sites became methylated specifically, which may be an epigenetic sign for the galanin gene to start a differentiation programme. The same sequence was moderately methylated in somatic donor cell and, after transplanted into an enucleated oocyte by nuclear transfer (NT), came rapidly demethylated to a completion, and then, at the blastocyst stage, re-methylated at exactly the same CpG sites, as observed so in normal blastocysts. The precise recapitulation of normal methylation reprogramming and programming at the galanin gene sequence in bovine cloned embryos gives a cue for the potential of cloned embryo to superintend the epigenetic states of foreign genome, even after global demethylation.
着床前发育过程中DNA甲基化的变化非常动态,这使得该过程成为衡量克隆胚胎对其体细胞基因组进行重编程能力的最具吸引力的实验靶点。然而,一个弱点是着床前阶段关于基因组序列的信息很少,这些序列在全基因组去甲基化后显示出位点特异性的重新甲基化;这些序列(如果有的话)可能是一个前沿课题,用于测试早期克隆胚胎中重编程/编程过程究竟是如何重现的。在此,我们报道了在牛神经肽甘丙肽基因序列上发生的独特DNA甲基化变化。通过体外受精(IVF)获得的早期牛胚胎中的甘丙肽基因序列在桑椹胚阶段之前一直保持低甲基化状态。到囊胚期,某些CpG位点特异性地发生了甲基化,这可能是甘丙肽基因启动分化程序的一种表观遗传标志。该相同序列在体细胞供体细胞中呈中度甲基化,通过核移植(NT)移植到去核卵母细胞后,迅速完全去甲基化,然后在囊胚期,在与正常囊胚中观察到的完全相同的CpG位点重新甲基化。牛克隆胚胎中甘丙肽基因序列正常甲基化重编程和编程的精确重现提示了克隆胚胎即使在全基因组去甲基化后仍有潜力监控外源基因组的表观遗传状态。