Honda Shinnosuke, Miki Yuka, Miyamoto Yuya, Kawahara Yu, Tsukamoto Satoshi, Imai Hiroshi, Minami Naojiro
Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan.
Laboratory Animal and Genome Sciences Section, National Institute for Quantum and Radiological Science and Technology, Chiba 263-8555, Japan.
J Reprod Dev. 2018 Aug 20;64(4):297-301. doi: 10.1262/jrd.2018-024. Epub 2018 May 3.
Oog1, an oocyte-specific gene that encodes a protein of 425 amino acids, is present in five copies on mouse chromosomes 4 and 12. In mouse oocytes, Oog1 mRNA expression begins at embryonic day 15.5 and almost disappears by the late two-cell stage. Meanwhile, OOG1 protein is detectable in oocytes in ovarian cysts and disappears by the four-cell stage; the protein is transported to the nucleus in late one-cell to early two-cell stage embryos. In this study, we examined the role of Oog1 during oogenesis in mice. Oog1 RNAi-transgenic mice were generated by expressing double-stranded hairpin Oog1 RNA, which is processed into siRNAs targeting Oog1 mRNA. Quantitative RT-PCR revealed that the amount of Oog1 mRNA was dramatically reduced in oocytes obtained from Oog1-knockdown mice, whereas the abundance of spermatogenesis-associated transcripts (Klhl10, Tekt2, Tdrd6, and Tnp2) was increased in Oog1 knockdown ovaries. Tdrd6 is involved in the formation of the chromatoid body, Tnp2 contributes to the formation of sperm heads, Tekt2 is required for the formation of ciliary and flagellar microtubules, and Klhl10 plays a key role in the elongated sperm differentiation. These results indicate that Oog1 down-regulates the expression of spermatogenesis-associated genes in female germ cells, allowing them to develop normally into oocytes.
Oog1是一种卵母细胞特异性基因,编码一个由425个氨基酸组成的蛋白质,在小鼠4号和12号染色体上有五个拷贝。在小鼠卵母细胞中,Oog1 mRNA表达始于胚胎第15.5天,到双细胞晚期几乎消失。与此同时,OOG1蛋白在卵巢囊肿的卵母细胞中可检测到,到四细胞期消失;该蛋白在单细胞晚期到双细胞早期胚胎中被转运到细胞核。在本研究中,我们研究了Oog1在小鼠卵子发生过程中的作用。通过表达双链发夹Oog1 RNA生成Oog1 RNAi转基因小鼠,该RNA被加工成靶向Oog1 mRNA的小干扰RNA。定量逆转录-聚合酶链反应显示,从Oog1基因敲低小鼠获得的卵母细胞中Oog1 mRNA的量显著减少,而在Oog1基因敲低的卵巢中,精子发生相关转录本(Klhl10、Tekt2、Tdrd6和Tnp2)的丰度增加。Tdrd6参与类染色质体的形成,Tnp2有助于精子头部的形成,Tekt2是纤毛和鞭毛微管形成所必需的,而Klhl10在精子延长分化中起关键作用。这些结果表明,Oog1下调雌性生殖细胞中精子发生相关基因的表达,使其能够正常发育成卵母细胞。