受精前後のストレス応答におけるエピジェネティック制御因子の役割と疾患発症への影響

• Project Area: The germline: its developmental cycle and epigenome network
• Project/Area Number: 21028005
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The University of Tokyo
• Principal Investigator: 栗原 裕基 東京大学, 大学院・医学系研究科, 教授 (20221947)
• Project Period (FY): 2009 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥3,200,000 (Direct Cost: ¥3,200,000); Fiscal Year 2010: ¥1,600,000 (Direct Cost: ¥1,600,000); Fiscal Year 2009: ¥1,600,000 (Direct Cost: ¥1,600,000)
• Keywords: 生殖細胞 / 初期胚発生 / エピジェネティック制御 / ストレス反応 / 病態生理

Research Abstract

本年度は、前年度に引き続いて受精前後のストレス応答におけるNAD+依存性脱アセチル化酵素Sirtuinの役割についての研究を行い、これを完成させて論文発表を行った(Kawamura et al., J. Clin. Invest.120;2817,2010)。受精卵ではすべてのSirtuin(Sirtl~7)が発現しており、このうちSirt3のmRNAおよびタンパクレベルが活性酸素種(ROS)刺激で増加することを示した。受精卵をSirtuin阻害薬で処理すると、発生の遅延や停止、及びミトコンドリア由来のROS産生の増加が認められた。siRNAインジェクションによるSirt3ノックダウンを行ったところ、Sirtuin阻害薬処理と同様に、胚盤胞形成率低下及びROS増加が認められた。Sirt3遺伝子ノックアウト胚でも体外受精後の培養により発生異常が生じた。抗酸化剤NAC処理や低酸素培養によりノックダウン胚の発生率が改善したことから、過剰ROS産生が発生異常の原因と考えられた。さらに、ノックダウン胚、ノックアウト胚においてP53及び下流遺伝子P21の発現増加、Nano9の発現低下が認められ、これらの変化がNACで抑制されたこと、Sirt3ノックダウンによる胚盤胞形成率低下がP53を同時にノックダウンすることにより改善したことから、p53シグナルの関与が示された。以上より、Sirt3は体外環境下における正常な初期発生進行にミトコンドリア機能調節を介して防御的に機能すると考えられた。本研究は、初期発生の制御機構解明に貢献するという生物学的意義をもつとともに、生殖補助医療における体外受精効率の影響因子の解明とその改善に向けた臨床研究にも貢献することが期待される。

Research Products

• [Journal Article] Calpain 6, a microtubule-stabilizing protein, regulates Rac1 activity and cell motility through interaction with GEF-H1. (2011)
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  • Journal Title: J.Cell Sci. Volume: 124 Pages: 1214-1223
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• [Journal Article] Endothelin receptor type-A expression defines a distinct cardiac subdomain within the heart field and is later implicated in chamber myocardium formation. (2011)
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  • Journal Title: Am.J.Physiol.Heart Circ.Physiol. Volume: 300(2)
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• [Journal Article] Dietary isoflavone increases insulin-like growth factor-I production, thereby promoting hair growth in mice. (2011)
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• [Journal Article] Sirt3 protects in vitro-fertilized mouse preimplantation embryos against oxidative stress-induced p53-mediated developmental arrest. (2010)
  • Author(s): Kawamura Y
  • Journal Title: J.Clin.Invest. Volume: 120 Pages: 2817-2828
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• [Journal Article] Endothelin receptor type-A expression defines a distinct cardiac subdomain within the heart field and is later implicated in chamber myocardium formation. (2010)
  • Author(s): Asai R
  • Journal Title: Development Volume: 137 Pages: 3823-3833
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• [Journal Article] Establishment of mice expressing EGFP in the placode-derived inner ear sensory cell lineage and FACS-array analysis focused on the regional specificity of the otocyst. (2010)
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• [Journal Article] Jaw muscularization requires Dlx expression by cranial neural crest cells. (2010)
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  • Journal Title: Proc.Natl.Acad.Sci.U.S.A. Volume: 107 Pages: 11441-11446
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• [Journal Article] Spatio-temporal dynamics of gene expression of the Edn1-Dlx5/6 pathway during development of the lower jaw. (2010)
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  • Journal Title: Genesis Volume: 48 Pages: 262-273
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• [Journal Article] Evolving maps in craniofacial development.Semin. (2010)
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  • Journal Title: Cell.Dev.Biol. Volume: 21 Pages: 301-308
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• [Journal Article] Pin1 associates with and induces translocation of CRTC2 to the cytosol, thereby suppressing CRE transcriptional activity. (2010)
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• [Journal Article] Cilostazol improves cognitive function in mice by increasing the production of insulin-like growth factor-I in the hippocampus. (2010)
  • Author(s): Zhao J
  • Journal Title: Neuropharmacology Volume: 58 Pages: 774-783
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• [Journal Article] Effects of topical application of alpha-D-glucosylglycerol on dermal levels of insulin-like growth factor-I in mice and on facial skin elasticity in humans. (2010)
  • Author(s): Harada N
  • Journal Title: Biosci.Biotechnol.Biochem. Volume: 74 Pages: 759-765
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• [Journal Article] Stimulation of FcγRI on primary sensory neurons increases insulin-like growth factor-I production, thereby reducing reperfusion-induced renal injury in mice. (2010)
  • Author(s): Harada N
  • Journal Title: J.Immunol. Volume: 185 Pages: 1303-1310
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• [Journal Article] In vitroライブイメージングによる血管新生機構の解明 (2010)
  • Author(s): 西山功一
  • Journal Title: 血管医学 Volume: 11(3) Pages: 77-84
• [Journal Article] エンドセリン受容体の多彩な機能---形態形成から病態発症まで (2010)
  • Author(s): 藤澤興
  • Journal Title: 医学の歩み Volume: 233(9) Pages: 846-850
• [Journal Article] Evolving maps in craniofacial development (2010)
  • Author(s): Gitton Y
  • Journal Title: Semin Cell Dev Biol. (in press 掲載確定(印刷中))
  • Peer Reviewed
• [Journal Article] Cilostazol improves cognitive function in mice by increasing the production of insulin-like growth factor-I in the hippocampus (2010)
  • Author(s): Zhao J
  • Journal Title: Neuropharmacology 58 Pages: 774-783
  • Peer Reviewed
• [Journal Article] Macrophage foam cell formation is augmented in serum from patients with diabetic angiopathy (2010)
  • Author(s): Cui X
  • Journal Title: Diabetes Res Clin Pract. 87 Pages: 57-63
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• [Journal Article] Inhibition of endothelial cell activation by bHLH protein E2-2 and its impairment of angiogenesis (2010)
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  • Journal Title: Blood (in press 掲載確定(印刷中))
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• [Journal Article] Transcriptional coactivator with PDZ-binding motif is essential for normal alveolarization in mice (2009)
  • Author(s): Mitani, A.
  • Journal Title: Am J Respir Crit Care Med. 180 Pages: 326-338
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• [Journal Article] The Hippo signaling pathway components Lats and Yap pattern Tead4 activity to distinguish mouse trophectoderm from inner cell mass (2009)
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  • Journal Title: Dev.Cell 16 Pages: 398-410
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• [Journal Article] A high-speed congenic strategy using first-wave male germ cells (2009)
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  • Journal Title: PLoS ONE 4
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• [Journal Article] Donepezil improves cognitive function in mice by increasing the production of insulin-like growth factor-I in the hippocampus (2009)
  • Author(s): Narimatsu N
  • Journal Title: J.Pharmacol.Exp.Ther. 330(1) Pages: 2-12
  • Peer Reviewed
• [Journal Article] Stimulation of sensory neurons improves cognitive function by promoting the hippocampal production of insulin-like growth factor-I in mice (2009)
  • Author(s): Harada, N.
  • Journal Title: Transl Res. 4 Pages: 90-102
  • Peer Reviewed
• [Presentation] Collective endothelial cell movements driving angiogenic morphogenesis (2011)
  • Author(s): 西山功一
  • Organizer: 第8回心血管幹細胞研究会
  • Place of Presentation: 品川プリンスホテル(東京都 港区)
  • Year and Date: 2011-01-15
• [Presentation] Coronary artery anomalies in Endothelin-1 and Endothelin A type receptor knock out mice. (2010)
  • Author(s): Yuichiro Arima
  • Organizer: 第18回日本血管生物医学会学術集会
  • Place of Presentation: 梅田スカイビル(大阪府 大阪市)
  • Year and Date: 2010-12-10
• [Presentation] Endothelin type-A receptor expression defines a distinct subpopulation within the first heart field contributing to chamber myocardium (2010)
  • Author(s): Rieko Asai
  • Organizer: 第33回日本分子生物学会・第83回日本生化学会合同年会
  • Place of Presentation: 神戸国際展示場(兵庫県 神戸市)
  • Year and Date: 2010-12-10
• [Presentation] Abnormal craniofacial morphogenesis induced by ectopic expression of Hoxgenes. (2010)
  • Author(s): Taro Kitazawa
  • Organizer: 第33回日本分子生物学会年会 第83回日本生化学会大会 合同大会
  • Place of Presentation: 神戸国際展示場(兵庫県 神戸市)
  • Year and Date: 2010-12-10
• [Presentation] Sirt3 protects in vitro-fertilized mouse preimplantation embryos against oxidative stress-induced p53-mediated developmental arrest (2010)
  • Author(s): Yumiko Kawamura
  • Organizer: 第33回 日本分子生物学会・第83回 日本生化学会合同年会
  • Place of Presentation: 神戸国際展示場(兵庫県 神戸市)
  • Year and Date: 2010-12-09
• [Presentation] Endothelin-1-mediated intracellular trafficking of endothelin receptor type A is regulated by the cytoplasmic C-termin us and causes the subtype-specific signaling in craniofacial development. (2010)
  • Author(s): Kou Fujisawa
  • Organizer: 第33回日本分子生物学会年会 第83回日本生化学会大会 合同大会
  • Place of Presentation: 神戸国際展示場(兵庫県 神戸市)
  • Year and Date: 2010-12-09
• [Presentation] In-depth analysis of angiogenic morphogenesis through time-lapse imaging and quantification (2010)
  • Author(s): 西山功一
  • Organizer: 第33回 日本分子生物学会・第83回 日本生化学会合同年会
  • Place of Presentation: 神戸ポートピアホテル(兵庫県 神戸市)
  • Year and Date: 2010-12-08
• [Presentation] Involvement of non-coding RNA in the Endotelin signaling in branchialarch development (2010)
  • Author(s): Yukiko Kurihara
  • Organizer: 第33回 日本分子生物学会・第83回 日本生化学会合同年会
  • Place of Presentation: 神戸国際展示場(兵庫県 神戸市)
  • Year and Date: 2010-12-08
• [Presentation] Transcriptional regulation of non-coding RNA Evf-2 in the Endothelin-1/Dlx5/Dlx6 pathway (2010)
  • Author(s): Yasunobu Uchijima
  • Organizer: 第33回 日本分子生物学会・第83回 日本生化学会合同年会
  • Place of Presentation: 神戸国際展示場(兵庫県 神戸市)
  • Year and Date: 2010-12-08
• [Presentation] Novel insight into angiogenesis : In-depth analysis through time-lapse imaging and quantification (2010)
  • Author(s): 西山功一
  • Organizer: 第18回日本血管生物医学会学術集会
  • Place of Presentation: 梅田スカイビル(大阪府 大阪市)
  • Year and Date: 2010-12-02
• [Presentation] A novel approach toward an understanding of angiogenesis using in vitro time-lapse live imaging (2010)
  • Author(s): 西山功一
  • Organizer: 第18回日本血管生物医学会学術集会
  • Place of Presentation: 梅田スカイビル(大阪府 大阪市)
  • Year and Date: 2010-12-01
• [Presentation] A newly identified phenomenon "Cell-mixing" during angiogenesis (2010)
  • Author(s): 西山功一
  • Organizer: 第18回日本血管生物医学会学術集会
  • Place of Presentation: 梅田スカイビル(大阪府 大阪市)
  • Year and Date: 2010-12-01
• [Presentation] Sirt3, a mitochondrial sirtuin, protects in vitro-fertilized mouse preimpl antation embryos against oxidative stress-induced p53-mediated developmental arrest (2010)
  • Author(s): Yumiko Kawamura
  • Organizer: International Symposium on "Epigenome Network, Development and Reprogramming of Germ Cells"
  • Place of Presentation: 九州大学医学部百年講堂(福岡県 福岡市)
  • Year and Date: 2010-11-23
• [Presentation] 胚形成を通してみるGPCRの機能解析--エンドセリン受容体のサブタイプ特異性- (2010)
  • Author(s): Kou Fujisawa
  • Organizer: 第7回東京呼吸器リサーチフォーラム
  • Place of Presentation: 帝人本社(東京都 千代田区)
  • Year and Date: 2010-11-06
• [Presentation] マウスにおける冠動脈の発生とその異常 (2010)
  • Author(s): Yuichiro Arima
  • Organizer: 第9回心臓血管発生研究会
  • Place of Presentation: シェラトン・グランデ・トーキョーベイ・ホテル(千葉県 浦安市)
  • Year and Date: 2010-07-10
• [Presentation] エンドセリンA受容体の発現による一次心臓領域におけるchamber myocardium寄与細胞系譜の同定 (2010)
  • Author(s): 淺井理恵子
  • Organizer: 第9回 心臓血管発生研究会
  • Place of Presentation: シェラトン・グランデ・トーキョーベイ・ホテル(千葉県 浦安市)
  • Year and Date: 2010-07-10
• [Presentation] Endothelin type-A receptor expression defines a distinct subdomain within the crescent-forming heart field contributing to chamber myocardium (2010)
  • Author(s): Rieko Asai
  • Organizer: Weinstein 2010 cardiovascular development conference
  • Place of Presentation: Royal Tropical Institute (AMSTERDAM, The Netherlands)
  • Year and Date: 2010-05-21
• [Presentation] Non-cording RNA regulation of endothelin signaling in branchial arch development (2010)
  • Author(s): Yukiko Kurihara
  • Organizer: Gordon Research Conference Craniofacial Morphogenesis & Tissue Regeneration
  • Place of Presentation: Il Ciocco Hotel & Resort (Lucca, Italy)
  • Year and Date: 2010-04-13
• [Presentation] Subtype-specific domain functions of endothelin receptors in craniofacial development. (2010)
  • Author(s): Kou Fujisawa
  • Organizer: Gordon Research Conference Craniofacial Morphog enesis & Tissue Regeneration
  • Place of Presentation: Il Ciocco Hotel & Resort (Lucca, Italy)
  • Year and Date: 2010-04-12
• [Presentation] Role of non-coding RNA Evf2 in the Endothelin-1/Dlx5/Dlx6 pathway regulation jaw morphogenesis (2009)
  • Author(s): 内島泰信
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜、パシフィコ横浜
  • Year and Date: 2009-12-10
• [Presentation] In vivo analysis of subtype-specific domain functions of endothelin receptors in craniofacial development (2009)
  • Author(s): 藤澤興
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜、パシフィコ横浜
  • Year and Date: 2009-12-10
• [Presentation] 活性中心欠失型カルパインによる低分子量Gタンパク質Rac1の活性制御機構の同定 (2009)
  • Author(s): 礪波一夫
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜、パシフィコ横浜
  • Year and Date: 2009-12-10
• [Presentation] Endothelin type-A receptor expression defines a distinct subdomain within the first heart field contributing to chamber myocardium (2009)
  • Author(s): 淺井理恵子
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜、パシフィコ横浜
  • Year and Date: 2009-12-09
• [Presentation] 活性中心欠失型カルパインによる新しい細胞骨格制御機構の同定 (2009)
  • Author(s): 礪波一夫
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸、神戸ポートアイラント
  • Year and Date: 2009-10-22
• [Presentation] 顎顔面の形態形成におけるエンドセリン受容体の in vivo ドメイン機能解析 (2009)
  • Author(s): 藤澤興
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸、神戸ポートアイランド
  • Year and Date: 2009-10-22
• [Presentation] Id1-mediated dynamic regulation of Notch signaling in vascular endothelial cells during angiogenesis (2009)
  • Author(s): 西山功一
  • Organizer: 第17会日本血管生物医学会大会
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2009-10-09
• [Presentation] in vitro ライブイメージングによる血管新生機構の解明 (2009)
  • Author(s): 西山功一
  • Organizer: 第17会日本血管生物医学会大会
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2009-10-08
• [Presentation] Modular analysis of angiogenic cell movement by real-time live imaging (2009)
  • Author(s): 有馬聡
  • Organizer: 第17会日本血管生物医学会大会
  • Place of Presentation: 東京、東京大学
  • Year and Date: 2009-10-08
• [Presentation] Functional analysis of subtype-specific domains of endothelin receptor type A in craniofacial development (2009)
  • Author(s): 藤澤興
  • Organizer: 第42回日本発生生物学会大会
  • Place of Presentation: 新潟、朱鷺メッセ
  • Year and Date: 2009-05-31
• [Presentation] Endothelin type-A receptor expression defines a distinct subpopulation within the first heart field contributing to chamber myocardium (2009)
  • Author(s): 淺井理恵子
  • Organizer: 第42回日本発生生物学会大会
  • Place of Presentation: 新潟、朱鷺メッセ
• [Presentation] Blood vessel formation as a stage of coordinated morphogenesis-Analysis using in vitro real-time imaging (2009)
  • Author(s): 西山功一
  • Organizer: 日本数理生物学会第20回年会
  • Place of Presentation: 東京、東京大学
• [Presentation] Id1 may control the specification and the timing of Notch signaling in vascular endothelial cells during angiogenesis (2009)
  • Author(s): 西山功一
  • Organizer: 第82会日本生化学会大会
  • Place of Presentation: 神戸、神戸ポートアイランド
• [Presentation] Id1 regulates angiogenesis by determining the specification and the timing of Notch signaling (2009)
  • Author(s): 西山功一
  • Organizer: American Heart Association, scientific session
  • Place of Presentation: Orland, USA
• [Remarks]
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• [Remarks]
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