Affiliated Hospital of Shaanxi University of Chinese Medicine, Shaanxi University of Chinese Medicine, Xianyang, Shaanxi, China.
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, China.
Cell Reprogram. 2021 Apr;23(2):99-107. doi: 10.1089/cell.2020.0079.
Zygotic epigenetic reprogramming is the major initial event in embryo development to acquire a totipotent potential. However, the patterns of epigenetic modifications in bovine zygote were not well clarified, especially in the first cell cycle of bovine somatic cell nuclear transfer (SCNT) embryos. This study was conducted to examine the patterns of DNA methylation (5-methylcytosine [5mc] and 5-hydroxymethylcytosine [5hmc]) and histone H3 lysine 9 methylation (H3K9m2 and H3K9m3) in the first cell cycle of bovine fertilization (IVF) and SCNT embryos. In bovine zygotic development, the 5mc in the paternal pronucleus (pPN) undergoes partial demethylation from PN1 to PN3, and remethylation from PN4 to PN5, while 5hmc exhibits absolutely different patterns. The 5mc in SCNT embryos underwent much more dramatic demethylation and much earlier methylation compared with their IVF counterparts, while 5hmc stayed stable from PN1 to PN4, and significantly increased at PN5, which made significantly higher level of 5mc and 5hmc at the end of the first cell cycle in SCNT embryos. Different H3K9m2 and H3K9m3 patterns were also observed between IVF and SCNT embryos. H3K9m2 and H3K9m3 asymmetrically distributed in parental genomes in IVF zygote, highly present in the maternal pronucleus, whereas faintly stained in the pPN. H3K9m2 and H3K9m3 in the somatic cell genome were gradually demethylated from PN1-PN4, and significantly increased at the end of the first cell cycle. dioxygenase was highly present in the first cell cycle of embryos compared with and . Our results showed that SCNT embryos underwent aberrant epigenetic reprogramming in the first cell cycle; much more dramatic demethylation and significant higher remethylation were observed compared with IVF counterparts.
合子的表观遗传重编程是胚胎发育获得全能性潜能的主要初始事件。然而,牛合子中的表观遗传修饰模式尚未得到很好的阐明,尤其是在牛体细胞核移植(SCNT)胚胎的第一个细胞周期中。本研究旨在研究牛受精(IVF)和 SCNT 胚胎第一个细胞周期中 DNA 甲基化(5-甲基胞嘧啶[5mc]和 5-羟甲基胞嘧啶[5hmc])和组蛋白 H3 赖氨酸 9 甲基化(H3K9m2 和 H3K9m3)的模式。在牛合子发育过程中,从 PN1 到 PN3,父本原核(pPN)中的 5mc 经历部分去甲基化,从 PN4 到 PN5 重新甲基化,而 5hmc 则表现出完全不同的模式。与 IVF 胚胎相比,SCNT 胚胎的 5mc 经历了更为剧烈的去甲基化和更早的甲基化,而 5hmc 则从 PN1 到 PN4 保持稳定,并在 PN5 显著增加,这使得 SCNT 胚胎在第一个细胞周期结束时的 5mc 和 5hmc 水平显著升高。IVF 和 SCNT 胚胎之间也观察到不同的 H3K9m2 和 H3K9m3 模式。在 IVF 合子中,H3K9m2 和 H3K9m3 在亲代基因组中不对称分布,在母本原核中高度存在,而在 pPN 中则微弱染色。从 PN1-PN4,SCNT 胚胎中体细胞基因组中的 H3K9m2 和 H3K9m3 逐渐去甲基化,并在第一个细胞周期结束时显著增加。与 和 相比,在胚胎的第一个细胞周期中高度存在。我们的研究结果表明,SCNT 胚胎在第一个细胞周期中经历了异常的表观遗传重编程;与 IVF 胚胎相比,观察到更为剧烈的去甲基化和显著更高的再甲基化。