Animal Genetic Resources Station, National Institute of Animal Science, RDA, Namwon, Korea.
Mol Reprod Dev. 2013 Mar;80(3):212-22. doi: 10.1002/mrd.22153. Epub 2013 Feb 28.
DNA methyltransferase1o (Dnmt1o), which is specific to oocyte and preimplantation embryo, plays a role in maintaining DNA methylation in mammalian cells. Here, we investigated the methylation status of CpGs sites in the Dnmt1o 5'-flanking region in germ cells at different stages of oogenesis or spermatogenesis. The methylation levels of the CpG sites at the 5'-flanking regions were hypermethylated in growing oocytes of all follicular stages, while the oocytes in meiotic metaphase II (MII) were demethylated. The methylation pattern within the CpGs sites in the 5'-flanking region, however, was dramatically changed during spermatogenesis. We observed that there was significant non-CpG methylation both in MII oocytes and spermatocytes. Although a low methylation level in non-CpG sites was observed in primary and secondary oocytes, the CpA site of position 25 and CpT site of position 29 within the no-CpG region in the 5'-flanking region of Dnmt1o was highly methylated in MII oocytes. During spermatogenesis, the low degree of methylation at CpG sites in spermatocytes increased to a higher degree in sperm, while the high ratio of methylation in non-CpG sites in spermatocytes decreased. Together, germ cells showed inverted methylation patterns between CpG and non-CpG sites in the Dnmt1o 5'-upstream region, and the methylation pattern during oogenesis did not drastically change, remaining generally hypomethylated at the MII stage.
DNA 甲基转移酶 1o(Dnmt1o)是卵母细胞和植入前胚胎特异性的,在哺乳动物细胞中维持 DNA 甲基化中发挥作用。在这里,我们研究了在卵母细胞发生或精子发生的不同阶段的生殖细胞中 Dnmt1o 5'-侧翼区的 CpG 位点的甲基化状态。在所有卵泡阶段的生长卵母细胞中,5'-侧翼区的 CpG 位点的甲基化水平呈高度甲基化,而处于减数分裂中期 II(MII)的卵母细胞则呈去甲基化。然而,在精子发生过程中,5'-侧翼区的 CpG 位点内的甲基化模式发生了剧烈变化。我们观察到,在 MII 卵母细胞和精母细胞中都存在显著的非 CpG 甲基化。尽管初级和次级卵母细胞中观察到非 CpG 位点的低甲基化水平,但 Dnmt1o 5'-侧翼区非 CpG 区域的位置 25 的 CpA 位点和位置 29 的 CpT 位点的甲基化水平在 MII 卵母细胞中非常高。在精子发生过程中,精母细胞中 CpG 位点的低甲基化程度在精子中增加到更高程度,而精母细胞中非 CpG 位点的高甲基化比率降低。总之,生殖细胞在 Dnmt1o 5'-上游区域的 CpG 和非 CpG 位点之间显示出反转的甲基化模式,并且卵母细胞发生过程中的甲基化模式没有剧烈变化,在 MII 阶段仍保持普遍低甲基化。