Dynamics of 5-hydroxymethylcytosine during mouse spermatogenesis
Dynamics of 5-hydroxymethylcytosine during mouse spermatogenesis
复制标题
5-羟甲基胞嘧啶在小鼠精子发生过程中的动态
DOI：
10.1038/ncomms2995
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发表时间：
2013-06-01
影响因子：
16.6
通讯作者：
Tang, Fuchou
中科院分区：
文献类型：
--
作者：
Gan, Haiyun;Wen, Lu;Tang, Fuchou
关于DNA甲基化模式在哺乳动物精子发生过程中是如何变化的，人们知之甚少。5hmC是一种稳定的DNA去甲基化中间体，在哺乳动物基因组中具有潜在的调控功能。然而，它在生殖细胞中的全球模式尚未得到解决。在这里，我们首先用液相色谱-串联质谱仪对连续八种类型的小鼠生精细胞中的5HmC进行了绝对定量，然后使用我们的化学标记和富集法结合深度测序的方法绘制了它在不同基因组区域的分布。我们发现，5hmC在与蛋白质编码基因、piRNA前体基因和重复元件的表达调控相关的基因组区域有差异定位和动态变化。此外，5hmC含量与RNA-seq定量的各种转录本的水平相关。这些结果表明，小鼠基因组中5hmC的高度有序变化对生精细胞的分化具有潜在的关键作用。
Little is known about how patterns of DNA methylation change during mammalian spermatogenesis. 5hmC has been recognized as a stable intermediate of DNA demethylation with potential regulatory functions in the mammalian genome. However, its global pattern in germ cells has yet to be addressed. Here, we first conducted absolute quantification of 5hmC in eight consecutive types of mouse spermatogenic cells using liquid chromatography-tandem mass spectrometry, and then mapped its distributions in various genomic regions using our chemical labeling and enrichment method coupled with deep sequencing. We found that 5hmC mapped differentially to and changed dynamically in genomic regions related to expression regulation of protein-coding genes, piRNA precursor genes and repetitive elements. Moreover, 5hmC content correlated with the levels of various transcripts quantified by RNA-seq. These results suggest that the highly ordered alterations of 5hmC in the mouse genome are potentially crucial for the differentiation of spermatogenic cells.