生殖細胞の成立と分化における分子基盤

• Project/Area Number: 15080206
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Osaka University
• Principal Investigator: 仲野 徹 Osaka University, 生命機能研究科, 教授 (00172370)
• Co-Investigator(Kenkyū-buntansha): 木村 透 大阪大学, 医学系研究科, 准教授 (50280962)
• Project Period (FY): 2003 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥114,600,000 (Direct Cost: ¥114,600,000); Fiscal Year 2007: ¥23,400,000 (Direct Cost: ¥23,400,000); Fiscal Year 2006: ¥23,700,000 (Direct Cost: ¥23,700,000); Fiscal Year 2005: ¥22,500,000 (Direct Cost: ¥22,500,000); Fiscal Year 2004: ¥22,500,000 (Direct Cost: ¥22,500,000); Fiscal Year 2003: ¥22,500,000 (Direct Cost: ¥22,500,000)
• Keywords: エピジェネヒィック制御 / DNAメチル化 / 遺伝子発現制御 / 初期胚 / レトロトランスポゾン / インプリンディング / 幹細胞 / 精子形成 / small RNA / インプリンティング / 生殖細胞 / 細胞文化 / シグナル伝達 / 遺伝子操作 / 細胞分化 / mili / RNAi / 遺伝子ターゲティングマウス / PTEN / germ cell less / 脱分化 / 胚性生殖細胞

Research Abstract

PGC7/Stellaは、初期胚、始原生殖細胞、卵細胞で特異的に発現し、受精後に細胞内局在が細胞質から核へと変化する。また、遺伝子改変マウスを用いた解析から、PGC7/Stellaは、卵子に存在する初期発生に必要な「母性因子」であることが明らかにされている。我々は、PGC7/Stellaの機能を解析するために、結合因子の探索を行い、タンパク質の核内輸送に関与するRanBP5(Rallbinding protein 5)を同定した。培養細胞を用いた実験から、RanBP5はPGC7/Stellaの核移行を促進することが明らかになった。次に、RanBPSとエストロゲン受容体のリガンド結合ドメインを融合させることにより、核に移行することができない細胞質局在型RanBP5を作製した。細胞質局在型RanBPSを受精卵に発現させると、PGC7/Stellaの核移行を阻害しただけではなく、PGC7/Stellaを欠損する卵子とほぼ同様の発生異常を示した。この発生異常は、核局在型PGC7/Stellaを同時に発現させることにより正常化することができた。
PGC7/Stellaが受精直後の核内で機能することから、受精卵おけるゲノムのメチル化状態を検討した。その結果、PGC7/Stella欠損の受精卵において、DNA複製が開始される前に雌性ゲノムの脱メチル化が生じていることが明らかとなった。また、PGC7/Stella欠損の受精卵において、いくつかのインプリント遺伝子のメチル化が低下していた。以上のことから、PGC7/Stellaは受精卵におけるエピジェネティック不均等性の成立および初期発生におけるインプリントの維持という、ゲノムの不均等性維持に重要な機能を有することが明らかとなった。

Research Products

• [Journal Article] Akt signaling promotes derivation of embryonic germ cells from primordial germ cells (2008)
  • Author(s): Kimura, T・Tomooka, M・Yamano, N, 3名, Kanai, Y・Nakano, T
  • Journal Title: Development (In press)
  • NAID: 120004837399
• [Journal Article] Differential context-dependent effects of FOG-1 on mast cell development and defferentiation (2008)
  • Author(s): Sugiyama, D・Tanaka, M・Kitajima, K, 3名, Yamatodani, A・Nakano, T
  • Journal Title: Blood (In press)
• [Journal Article] PGC7/Stella protects against DNA demethyaltion in early embryogenesis (2007)
  • Author(s): Nakamura, T・Arai, Y・Umehara, H, 8名, Shiota, K・Nakano, T
  • Journal Title: Nature Cell Biol 9 Pages: 64-71
• [Journal Article] Akt activation induces epidermal hyperplasia and proliferation epidermal progenitors (2007)
  • Author(s): Murayama, K・Kimura, T・Tarutani, M, 5名, Katayama, I・Nakano, T
  • Journal Title: Oncogene 26 Pages: 4882-4888
• [Journal Article] Pten/PI3K pathway governs the homeostasis of Valpha14iNKT cells (2007)
  • Author(s): Kishimoto, H・Ohdeki, T, 11名, Nakano, T・Koyasu, S・Sasaki, T・Suzuki
  • Journal Title: Blood 109 Pages: 3316-3324
• [Journal Article] Efficient derivation of embryonic stem cells by inhibition of glycogen synthasekinase-3 (2007)
  • Author(s): Umehara, H・Kimura, T・Ohtsuka, S, 4名, Niwa, H・Nakano, T
  • Journal Title: Stem Cells 25 Pages: 2705-2711
• [Journal Article] Differential Effects of GATA-1 on Proliferation and Differentiation of Erythroid Lineage Cells (2006)
  • Author(s): Zheng J, Kitajima K, Sakai E, Kimura T, Minegishi N, Yamamoto M, Nakano T
  • Journal Title: Blood 107 Pages: 520-527
• [Journal Article] Activation of Akt Signaling is Sufficient to Maintain Pluripotency in Mouse and Primate Embryonic Stem Cells (2006)
  • Author(s): Watanabe S, Umehara H, Murayama K, Okabe M, Kimura T, Nakano T
  • Journal Title: Oncogene 25 Pages: 2697-2707
• [Journal Article] Redirecting Differentiation of Hematopoietic Progenitors by a Transcription Factor GATA-2 (2006)
  • Author(s): Kitajima K, Tanaka M, Yen H, Sato A, Umehara H, Sakai E, Nakano T
  • Journal Title: Blood 107 Pages: 1857-1863
• [Journal Article] Multipotential differentiation ability of GATA-1 null erythroid committed cells (2006)
  • Author(s): Kitajima K, Zheng J, Yen H, Sugiyama D, Nakano T
  • Journal Title: Genes ＆ Development 20 Pages: 654-659
• [Journal Article] A novel class of small RNAs bind to MILI protein in mouse testes (2006)
  • Author(s): Aravin A, Gaidatzis D, (8名), Nakano T, (6名), Tuschl T
  • Journal Title: Nature 442 Pages: 203-207
• [Journal Article] Characterization of the piRNA complex from rat testes (2006)
  • Author(s): Lau NC, (3名), Nakano T, Bartel DP, Kingston RE
  • Journal Title: Science 313 Pages: 363-367
• [Journal Article] Redirecting Differentiation of Hematopoietic Progenitors by a Tanscription Factor GATA-2 (2006)
  • Author(s): Kitajima K, Tanaka M, Yen H, Sato A, Sugiyama D, Umehara H, Sakai E, Nakano T
  • Journal Title: Blood 107 Pages: 1857-1863
• [Journal Article] Differential Effects of GATA-1 on Proliferation and Differentiation of Erythroid Lineage Cells (2006)
  • Author(s): Zheng J, Kitajima J, Sakai E, Kimura T, Minegishi N, Yamamoto M, Nakano T
  • Journal Title: Blood 107 Pages: 520-527
• [Journal Article] The PTEN/PI3K pathway governs normal vascular development and tumor angiogenesis (2005)
  • Author(s): Hamada K, Sasaki T, (10名), Mak TW, Nakano T, Suzuki A
  • Journal Title: Genes & Development 19 Pages: 2054-2065
• [Journal Article] Involvement of connective tissue type mast cells in Th1 immune responses via Stat4 expression (2005)
  • Author(s): Kataoka TR, Komazawa N, Morii E, Ohboki K, Nakano T
  • Journal Title: Blood 105 Pages: 1016-1020
• [Journal Article] Reduced expression of IL-12 receptor β and IL-18 receptor □ genes in natural killer cells and macrophages derived from B6^<-mi/mi> mice (2005)
  • Author(s): Kataoka TR, Komazawa N, Morii E, Ohboki H, Nakano T
  • Journal Title: Lab Invest 85 Pages: 146-153
• [Journal Article] mili, a mammalian member of piwi family gene, is essential for spermatogenesis (2004)
  • Author(s): Kuramochi-Miyagawa K, Kimura T, (10 authors), Nakano T
  • Journal Title: Development 131 Pages: 839-849
  • NAID: 120004837540
• [Journal Article] Hepatocyte-specific Pten deficiency results in steatohepatitis and hepatocellular carcinomas (2004)
  • Author(s): Horie Y, Suzuki A, (10 authors), Mak TW, Nakano T
  • Journal Title: J Clin Invest 113 Pages: 1174-1783
• [Journal Article] Differentiation Status Dependent Function of FOG-1 (2004)
  • Author(s): Tanaka T, Zheng J, Kitajima K, Kita K, Yoshikawa H, Nakano T
  • Journal Title: Genes to Cells 9 Pages: 1213-1226
• [Journal Article] Keratinocyte-specific Pten deficiency results in epidermal hyperplasia, accelerated hair follicle morphogenesis and tumor formation (2003)
  • Author(s): Suzuki A, Itami A, (8 authors), Nakano T
  • Journal Title: Cancer Res 63 Pages: 674-681
• [Journal Article] Multipotential differentiation ablity of GATA-1 null erythroid committed cells
  • Author(s): Kitajima K, Zheng J, Yen H, Sugiyama D, Nakano T
  • Journal Title: Genes & Development 2006(In press)
• [Journal Article] Activation of Akt Signaling is Sufficient to Maintain Pluripotency in Mouse and Primate Embryonic Stem Cells
  • Author(s): Watanabe S, Umehara H, Murayama K, Okabe M, Kimura T, Nakano T
  • Journal Title: Oncogene 2006(In press)
• [Book] Testicular Teratomas: Back to Pluripotent Stem Cells in Developmental Biology of Neoplastic Growth (2005)
  • Author(s): Kimura T, Murayama K, Nakamura T, Watanabe S, Umehara H, Tomooka M, Nakano T
  • Total Pages: 15
  • Publisher: Springer, Berlin
• [Book] バイオテクノロジーのための基礎分子生物学 (2004)
  • Author(s): 仲野 透(共著)
  • Total Pages: 201
  • Publisher: 化学同人
• [Publications] Suzuki A, Kaisho T, (6 authors), Nakano T.: "Critical roles of Pten in B cell homeostasis and immunoglobulin class switch recombination"J Exp Med. 197. 657-667 (2003)
• [Publications] Kimura T, Suzuki A, (7 authors), Nakano T: "Conditional loss of PTEN leads to testicular teratoma and enhances embryonic germ cell production"Development. 130. 1691-1700 (2003)
• [Publications] Kimura T, Ito C, (11 authors), Nakano T.: "Mouse Germ Cell-Less as an essential component for nuclear integrity"Mol Cell Biol. 23. 1304-1315 (2003)
• [Publications] Suzuki A, Itami A, (8 authors), Nakano T: "Keratinocyte-specific Pten deficiency results in epidermal hyperplasia, accelerated hair follicle morphogenesis and tumor formation"Cancer Res. 63. 674-681 (2003)
• [Publications] Nakano T: "Hematopoietic stem cells : generation and manipulation"Trends Immunol. 24. 589-594 (2003)
• [Publications] Kuramochi-Miyagawa K, Kimura T, (10 authors), Nakano T: "mili, a mammalian member of piwi family gene, is essential for spermatogenesis"Development. 131. 839-849 (2004)
• [Publications] 仲野 徹: "幹細胞とクローン"羊土社. 115 (2003)