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Functional analysis of Oog1, an oocyte-specific gene, in mouse embryo
https://repo.qst.go.jp/records/61998
https://repo.qst.go.jp/records/619988d696497-100b-428e-a6e1-f6648c4fc283
| Item type | 会議発表用資料 / Presentation(1) | |||||
|---|---|---|---|---|---|---|
| 公開日 | 2007-01-23 | |||||
| タイトル | ||||||
| タイトル | Functional analysis of Oog1, an oocyte-specific gene, in mouse embryo | |||||
| 言語 | ||||||
| 言語 | eng | |||||
| 資源タイプ | ||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_c94f | |||||
| 資源タイプ | conference object | |||||
| アクセス権 | ||||||
| アクセス権 | metadata only access | |||||
| アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
| 著者 |
Tukamoto, Satoshi
× Tukamoto, Satoshi× Kito, Seiji× Oota, Yuki× Aizawa, Akira× Imai, Hiroshi× Minami, Naojiro× et.al× 鬼頭 靖司× 太田 有紀× 南 直治郎 |
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| 抄録 | ||||||
| 内容記述タイプ | Abstract | |||||
| 内容記述 | Maternal effect, which can refer to the dependence of early embryonic development on maternal products, is common in both animal and plant kingdom. Nine mammalian maternal-effect genes, such as Mater, Hsf1, Dnmt1o, Pms2, Zar1, Npm2, stella, Zfp36l2 and Basonuclin, have been identified recently, largely by gene-knockout strategies. In the mouse, zygotic genome activation (ZGA) occurs at the late 1-cell to early 2-cell stages and is a critical event that governs the maternal-to-zygotic transition that is indispensable for further development. Some of the maternal effect genes may be involved in the ZGA. However, the molecular mechanisms of maternal-effect genes have not been well understood. We previously identified an oocyte-specific gene, Oogenesin 1 (Oog1) expressed throughout oogenesis and early embryogenesis. The primary structure of Oog1 protein, deduced from the full-length cDNA (1387 bp), is composed of 326 amino acids (37 kDa) containing a leucine zipper structure and a leucine-rich repeat (LRR). Oog1 mRNA is present specifically in oocytes and preimplantation embryos, and the protein was detected in oocytes until the 4-cell stage. Interestingly, Oog1 protein was found to be localized in nuclei at the late 1-cell and the early 2-cell stages. Oog1 was recently shown to be a member of the Oogenesin family, whose members (Oog1, 2, 3, 4) are clustered on mouse chromosome 4 and these proteins belong to the LRR superfamily which is involved in protein-protein interactions. We have also reported that Oog1 interacts with Ras effectors (e.g., RalGDS and Rasa4) and binds to Ras in a GTP-dependent manner. These results suggested that Oog1 is one of the Ras-mediated signaling proteins in early embryogenesis. However, the precise function of Oog1 in oogenesis and/or early embryogenesis is still unclear. In the present study, to examine Oog1 function in preimplantation embryos, we carried out the RNAi-mediated experiments using Oog1-hairpin. The effect of Oog1-hairpin dsRNA on the reduction of Oog1 mRNA level were confirmed by microinjection of the transcribed Oog1-hairpin dsRNA fused with EGFP into the cytoplasm of GV-oocytes. Following microinjection, the expression of the EGFP was detected after 4-5 h by fluorescent microscopy and the amount of Oog1 mRNA in oocytes with EGFP signals was confirmed after 20 h by RT-PCR. RT-PCR analysis showed that Oog1 transcript is dramatically reduced, although the abundance of Oog2, 3, and 4 transcripts, which are closely related homologues with Oog1, was not affected. These results suggest that the Oog1-hairpin dsRNA used in this experiment effectively and specifically reduced Oog1 mRNA. To investigate the Oog1 function in preimplantation development, we microinject the same RNA into MII-oocytes. Injected-oocytes were kept in culture medium until the EGFP signals were detected. The EGFP-positive oocytes were selected under fluorescent microscope and then in vitro fertilization (IVF) was performed. When these injected and fertilized embryos were analyzed morphologically until the blastocyst stages, it became apparent that no difference was observed in embryos injected with either Oog1-hairpin dsRNA or EGFP dsRNA as a control. In addition, we noted by western blot analysis that Oog1 protein was still present in embryos injected with Oog1-hairpin dsRNA. These results showed that knock-down of Oog1 mRNA by RNAi leads to an effective decrease in the mRNA, but not the protein. Generation of transgenic mice that express the Oog1-hairpin dsRNA during oogenesis will allow us to define the function of Oog1 during oogenesis and/or early embryogenesis in near future. |
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| 会議概要（会議名, 開催地, 会期, 主催者等） | ||||||
| 内容記述タイプ | Other | |||||
| 内容記述 | The 3rd Asian Reproductive Biotechnology Conference | |||||
| 発表年月日 | ||||||
| 日付 | 2006-12-03 | |||||
| 日付タイプ | Issued | |||||
