In situ alterations of DNA methylation were studied between 14 d postcoitum and 4 d postpartum in Sertoli cells and germ cells from mouse testis, using anti-5-methylcytosine antibodies. Compared to cultured fibroblasts, Sertoli cells display strongly methylated juxtacentromeric heterochromatin, but hypomethylated chromatids. Germ cells always possess hypomethylated heterochromatin, whereas their euchromatin passes from a demethylated to a strongly methylated status between days 16 and 17 postcoitum. This hypermethylation occurs in the absence of DNA replication, germ cells being blocked in the G(0)-G(1) phase from day 15 postcoitum to birth. The DNA hypermethylation of germ cells is maintained until birth and could be visualized on both chromatids of metaphase chromosomes at the first postpartum cell division. Subsequently, the DNA hypermethylation is lost semiconservatively, being replaced by a methylation pattern recalling the typical fibroblast pattern. These alterations of DNA methylation follow a strict chronology, are chromosome structure and cell-type dependent, and may underlie profound changes of genome function.

译文

使用anti-5-methylcytosine抗体研究了小鼠睾丸支持细胞和生殖细胞在产后14 d和产后4 d之间DNA甲基化的原位改变。与培养的成纤维细胞相比,Sertoli细胞显示出强烈的甲基化的间生异染色质,但低甲基化的染色单体。生殖细胞始终具有低甲基化的异染色质,而它们的常染色质在第16至17天之间从去甲基化状态转移到强烈甲基化状态。这种高甲基化发生在没有DNA复制的情况下,生殖细胞从出生后第15天到出生在G(0)-G(1) 阶段被阻断。生殖细胞的DNA高甲基化一直维持到出生,并且可以在产后第一次细胞分裂时在中期染色体的两个染色单体上观察到。随后,DNA超甲基化被半保守地丢失,被甲基化模式取代,使其回想起典型的成纤维细胞模式。DNA甲基化的这些改变遵循严格的时间顺序,是染色体结构和细胞类型依赖性的,并且可能是基因组功能深刻变化的基础。

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