Genes expressed specifically in oocytes may have an important role in the quality (developmental competence) of oocytes
| Project/Area Number |
19591911
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Keio University |
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Principal Investigator |
HAMATANI Toshio Keio University, 医学部, 講師 (60265882)
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| Co-Investigator(Kenkyū-buntansha) |
YOSHIMURA Yasunori 慶應義塾大学, 医学部, 教授 (10129736)
AKUTSU Hidenori 国立成育医療センター研究所, 生殖医療研究部, 室長 (50347225)
YAMADA Mitsutoshi 国立成育医療センター研究所, 生殖医療研究部, 研究員 (40383864)
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| Project Period (FY) |
2007 – 2009
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| Project Status |
Completed (Fiscal Year 2009)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2009: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2008: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2007: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
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| Keywords | 生殖医学 / 卵 / 胚性幹細胞 / 着床前期胚 / 遺伝子発現プロファイリング / RNAi / トランスジェニック・マウス |
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| Research Abstract |
To elucidate molecular mechanisms underlying oogenesis and oocyte quality, we first employed in silico analysis of global gene expression profiling data and an EST frequency database, and selected genes that are specifically expressed in oocytes and preimplantation embryos. We further studied the differences of gene expression profiles (1) between oocytes from aged mice and those from young mice, and (2) between post-ovulatory aged oocytes and fresh oocytes to extract genes that are important for oocyte quality. Finally, we focused on two oocyte-specific gene families (Nlrp [NACHT, leucine-rich repeat and PYD containing] genes and KRAB [Kruppel-associated box] zinc finger genes).
As results of quantitative RT-PCR and in situ hybridization assays, we confirmed oocyte-specific expression of the Nlrp and the KRAB genes at the follicle stages later than the primary-follicle stage. To analyze gene function of the Nlrp genes, we used the RNA interference approach : injection with siRNAs into
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mouse fertilized eggs. The knockdown of a Nlrp gene lead to cleavage arrest during preimplantation development. On the other hand, we are now in the process of generating knockout mice for an oocyte-specific KRAB zinc finger gene.
In addition, we study a preimplantation-stage-specific gene Hmgpi which is a putative chromosomal protein with two high-mobility-group boxes. Hmgpi is zygotically transcribed during zygotic genome activation, but not transcribed after implantation. The Hmgpi-encoded protein (HMGPI), first detected at the 4-cell stage, remains highly expressed in pre-implantation embryos. Interestingly, HMGPI is expressed in both the inner cell mass (ICM) and the trophectoderm, and translocated from cytoplasm to nuclei at the blastocyst stage, indicating differential spatial requirements before and after the blastocyst stage. siRNA (siHmgpi)-induced reduction of Hmgpi transcript levels caused developmental loss of preimplantation embryos and implantation failures. Furthermore, reduction of Hmgpi prevented blastocyst outgrowth leading to generation of embryonic stem cells. The siHmgpi-injected embryos also lost ICM and trophectoderm integrity, demarcated by reduced expressions of Oct4, Nanog and Cdx2. The findings implicated an important role for Hmgpi at the earliest stages of mammalian embryonic development. Less
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Report
(4 results)
Research Products
(32 results)
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[Journal Article]2009
Author(s)
浜谷敏生, 山田満稔, 吉村泰典
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Journal Title
よくわかる病態生理(監修:松尾理)第12巻婦人科疾患【構造と機能】女性生殖器の発育過程, 性周期発現と排卵の機序(編集:久保田俊郎)(日本医事新報社, 東京)
Related Report
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[Journal Article] What can you learn from gene expression profiling of mouse oocytes?2008
Author(s)
Hamatani T, Yamada M, Akutsu H, Kuji N, Mochimaru Y, Takano M, Toyoda M, Miyado M, Umezawa A, Kuji N, Yoshimura Y
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Journal Title
Reproduction 135(5)
Pages: 581-592
Related Report
Peer Reviewed
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[Journal Article] What can you learn from gene expression profiling of mouse oocytes?2008
Author(s)
Hamatani T, Yamada M, Akutsu H, Kuji N, Mochimaru Y, Takano M, Toyoda M, Miyado M, Umezawa A, Kuji N, Yoshimura Y.
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Journal Title
Reproduotion 135(5)
Pages: 581-592
Related Report
Peer Reviewed
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[Journal Article] What can you learn from gene expression profiling of mouse oocytes2008
Author(s)
Hamatani T, Yamada M, Akutsu H, Kuji N, Mochimaru Y, Takano M, Toyoda M, Miyado M, Umezawa A, Kuji N, Yoshimura Y.
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Journal Title
Reproduction 135(5)
Pages: 581-592
Related Report
Peer Reviewed
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