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The Promoter of the Oocyte-Specific Gene Oog1 Functions in Both Male and Female Meiotic Germ Cells in Transgenic Mice

机译:卵母细胞特异性基因Oog1的启动子在转基因小鼠的男性和女性减数分裂生殖细胞中均起作用

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摘要

Oog1 is an oocyte-specific gene whose expression is turned on in mouse oocytes at embryonic day (E) 15.5, concomitant with the time when most of the female germ cells stop proliferating and enter meiotic prophase. Here, we characterize the Oog1 promoter, and show that transgenic GFP reporter expression driven by the 2.7 kb and 3.9 kb regions upstream of the Oog1 transcription start site recapitulates the intrinsic Oog1 expression pattern. In addition, the 3.9 kb upstream region exhibits stronger transcriptional activity than does the 2.7 kb region, suggesting that regulatory functions might be conserved in the additional 1.2 kb region found within the 3.9 kb promoter. Interestingly, the longer promoter (3.9 kb) also showed strong activity in male germ cells, from late pachytene spermatocytes to elongated spermatids. This is likely due to the aberrant demethylation of two CpG sites in the proximal promoter region. One was highly methylated in the tissues in which GFP expression was suppressed, and another was completely demethylated only in Oog1pro3.9 male and female germ cells. These results suggest that aberrant demethylation of the proximal promoter region induced ectopic expression in male germ cells under the control of 3.9 kb Oog1 promoter. This is the first report indicating that sex-dependent gene expression is altered according to the length and the methylation status of the promoter region. Additionally, our results show that individual CpG sites are differentially methylated and play different roles in regulating promoter activity and gene transcription.
机译:Oog1是一种卵母细胞特异性基因,其表达在胚胎第(E)15.5天在小鼠卵母细胞中开启,同时伴随着大多数雌性生殖细胞停止增殖并进入减数分裂前期的时间。在这里,我们表征Oog1启动子,并表明由Oog1转录起始位点上游2.7 kb和3.9 kb区域驱动的转基因GFP报告基因表达了内在的Oog1表达模式。此外,3.9 kb的上游区域比2.7 kb的区域表现出更强的转录活性,这表明调节功能可能在3.9 kb的启动子中发现的其他1.2 kb区域中保守。有趣的是,较长的启动子(3.9 kb)在雄性生殖细胞中也表现出强大的活性,从后期的粗线精细胞到细长的精细胞。这可能是由于近端启动子区域中两个CpG位点异常脱甲基。一种在被抑制GFP表达的组织中高度甲基化,另一种仅在Oog1pro3.9雄性和雌性生殖细胞中完全脱甲基。这些结果表明在3.9 kb Oog1启动子的控制下,近端启动子区域的异常去甲基化诱导了雄性生殖细胞中的异位表达。这是第一个报道,表明性别依赖性基因表达根据启动子区域的长度和甲基化状态而改变。此外,我们的结果表明,单个CpG位点差异甲基化,并在调节启动子活性和基因转录中发挥不同作用。

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