Locus-Specific DNA Methylation Reprogramming During Early Porcine Embryogenesis
Locus-Specific DNA Methylation Reprogramming During Early Porcine Embryogenesis
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DOI：
10.1095/biolreprod.112.104471
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发表时间：
2013-02-01
影响因子：
3.6
通讯作者：
Prather, Randall S.
中科院分区：
文献类型：
--
作者：
Zhao, Ming-Tao;Rivera, Rocio M.;Prather, Randall S.
在哺乳动物早期胚胎发生过程中，受精后存在一个DNA去甲基化的高峰，着床前后存在一个DNA去甲基化的高峰。在大多数哺乳动物中，除了绵羊和兔子外，父本基因组经历主动DNA去甲基化，而母本基因组在受精后被动去甲基化。然而，新兴的全基因组DNA甲基化景观揭示了哺乳动物植入前胚胎中的调控和基因座特异性DNA甲基化重编程模式。在这里，我们优化了亚硫酸氢盐测序方案，以绘制配子，早期胚胎和体细胞组织中几个选定基因的碱基分辨率DNA甲基化谱。我们在早期猪胚胎中观察到基因座特异性DNA甲基化重编程。首先，一些多能性基因（POU 5 F1和NANOG）遵循典型的DNA去甲基化和再甲基化波，而SOX 2和CDX 2基因座的富含CpG的区域在整个发育过程中甲基化不足。第二，IGF 2/H19基因座中印记控制区的差异甲基化区域表现出差异DNA甲基化，其在猪早期胚胎中保持。第三，着丝粒重复元件在配子、早期胚胎和体细胞组织中保持中等的DNA甲基化水平。早期胚胎发生过程中DNA甲基化重编程的多样性被认为与DNA甲基化在转录调控、基因组稳定性和基因组印记等方面的多重功能有关。利用氧化亚硫酸氢盐测序等新技术鉴定5-羟甲基胞嘧啶，将进一步阐明猪胚胎发育过程中DNA甲基化重编程。
During early mammalian embryogenesis, there is a wave of DNA demethylation postfertilization and de novo methylation around implantation. The paternal genome undergoes active DNA demethylation, whereas the maternal genome is passively demethylated after fertilization in most mammals except for sheep and rabbits. However, the emerging genome-wide DNA methylation landscape has revealed a regulatory and locus-specific DNA methylation reprogramming pattern in mammalian preimplantation embryos. Here we optimized a bisulfite sequencing protocol to draw base-resolution DNA methylation profiles of several selected genes in gametes, early embryos, and somatic tissue. We observed locus-specific DNA methylation reprogramming in early porcine embryos. First, some pluripotency genes (POU5F1 and NANOG) followed a typical wave of DNA demethylation and remethylation, whereas CpG-rich regions of SOX2 and CDX2 loci were hypomethylated throughout development. Second, a differentially methylated region of an imprint control region in the IGF2/H19 locus exhibited differential DNA methylation which was maintained in porcine early embryos. Third, a centromeric repeat element retained a moderate DNA methylation level in gametes, early embryos, and somatic tissue. The diverse DNA methylation reprogramming during early embryogenesis is thought to be possibly associated with the multiple functions of DNA methylation in transcriptional regulation, genome stability and genomic imprinting. The latest technology such as oxidative bisulfite sequencing to identify 5-hydroxymethylcytosine will further clarify the DNA methylation reprogramming during porcine embryonic development.