Molecular mechanisms of hormone release revealed by live cell imaging analysis

• Project/Area Number: 18689008
• Research Category: Grant-in-Aid for Young Scientists (A)
• Allocation Type: Single-year Grants
• Research Field: General physiology
• Research Institution: The University of Tokyo (2007-2008); The Institute of Physical and Chemical Research (2006)
• Principal Investigator: TSUBOI Takashi The University of Tokyo, 大学院・総合文化研究科, 准教授 (80415231)
• Project Period (FY): 2006 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥27,690,000 (Direct Cost: ¥21,300,000、Indirect Cost: ¥6,390,000); Fiscal Year 2008: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000); Fiscal Year 2007: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000); Fiscal Year 2006: ¥17,160,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥3,960,000)
• Keywords: 可視化 / 生体分子 / 生理学 / 蛋白質 / 脳・神経 / 開口放出 / ホルモン分泌 / 全反射蛍光顕微鏡 / イメージング / 小胞輸送 / シナプトタグミン / 分泌顆粒輸送 / 低分子量GTP蛋白質 / Rab / ドッキング

Research Abstract

私達の身体は環境変化に対応するため、ホルモンを分泌することにより体内の恒常性を一定に保っている。この機構の破綻は、アレルギー、糖尿病等の種々の疾患に直結する。しかしホルモン分泌の詳細な分子メカニズムについては解明されていない。そこで、超高感度、超高解像度を持つ蛍光顕微鏡を新規開発し、内分泌細胞から起こるホルモン分泌反応を直接可視化計測することに成功した。そして、ホルモン分泌制御機構の詳細な分子メカニズムの一端を解明した。

Research Products

• [Journal Article] Neuroepithelial cells require fucosylated glycans to guide the migration of vagus motor neuron progenitors in the developing zebrafish hindbrain (2009)
  • Author(s): Ohata S, Kinoshita S, Aoki R, Tanaka H, Wada H, Tsuruoka-Kinoshita S, Tsuboi T, Watabe S, Okamoto H
  • Journal Title: Development (In press)
  • Peer Reviewed
• [Journal Article] Regulation of the intracellular distribution, cell surface expression, and protein levels of AMPA receptor GluR2 subunits by the monocarboxylate transporter MCT2 in neuronal cells (2009)
  • Author(s): Maekawa F, Tsuboi T, Pellerin L
  • Journal Title: Journal of Neurochemistry (In press)
  • Peer Reviewed
• [Journal Article] Molecular mechanism of attachment process of dense-core vesicles to the plasma membrane in neuroendocrine cells (2009)
  • Author(s): Tsuboi T
  • Journal Title: Neuroscience Research 63 Pages: 83-88
  • NAID: 10027499443
  • Peer Reviewed
• [Journal Article] Imaging a target of Ca^<2+> signalling: Dense core granule exocytosis viewed by total internal reflection fluorescence microscopy (2008)
  • Author(s): Ravier MA, Tsuboi T, Rutter GA
  • Journal Title: Methods 46 Pages: 233-238
  • Peer Reviewed
• [Journal Article] Molecular mechanism of docking of dense-core vesicle to the plasma membrane in neuroendocrine cells (2008)
  • Author(s): Tsuboi T
  • Journal Title: Medical Molecular Morphology 41(2) Pages: 68-75
  • Peer Reviewed
• [Journal Article] Rabphilin directly interacts with SNAP-25 and regulates the docking step of dense-core vesicle exocytosis in PC12 cells (2008)
  • Author(s): Tsuboi T
  • Journal Title: The Journal of Physiological Sciences 58 (Suppl.)
  • NAID: 130005449384
  • Peer Reviewed
• [Journal Article] Molecular mechanism of docking of dense-core vesicles to the plasma membrane in neuroendocrine cells (2008)
  • Author(s): Tsuboi T
  • Journal Title: Medical Molecular Morphology 41 Pages: 68-75
  • Peer Reviewed
• [Journal Article] Imaging a target of Ca2+ signalling: Dense core granule exocytosis viewed by totalinternal reflection fluorescence microscopy (2008)
  • Author(s): Ravier MA, Tsuboi T, Rutter GA
  • Journal Title: Methods 46 Pages: 233-238
  • Peer Reviewed
• [Journal Article] Rabphilin directly interacts with SNAP-25 and regulates the docking step of densecore vesicle exocytosis in PC12 cells (2008)
  • Author(s): Tsuboi T
  • Journal Title: The Journal of Physiological Sciences 58(suppl.)
• [Journal Article] The polybasic sequence in the C2B domain of rabphilin is required for the vesicle docking step in PC12 cells (2007)
  • Author(s): Tsuboi T, Kanno E, Fukuda M
  • Journal Title: Journal of Neurochemistry 100 Pages: 770-779
  • Peer Reviewed
• [Journal Article] Role for synaptotagmin VII in fusion pore dynamics revealed by total internal reflection fluorescence microscopy (2007)
  • Author(s): Tsuboi T, Fukuda M
  • Journal Title: Neuroscience Research 58 (suppl.1)
  • Peer Reviewed
• [Journal Article] Live cell imagingによる細胞活動の可視化解析 (2007)
  • Author(s): 坪井貴司
  • Journal Title: 日本比較内分泌学会ニュース 124 Pages: 45-53
  • NAID: 10018293579
• [Journal Article] Synaptotagmin VII modulates the kinetics of dense-core vesicle exocytosis in PC12 cells (2007)
  • Author(s): Tsuboi T, Fukuda M
  • Journal Title: Genes to Cells 12 Pages: 511-519
  • Peer Reviewed
• [Journal Article] The polybasic sequence in the C2B domain of rabphilin is required for the vesicle docking step in PC12 cells. (2007)
  • Author(s): Tsuboi T, 他2名
  • Journal Title: Journal of Neurochemistry 100 Pages: 770-779
  • Peer Reviewed
• [Journal Article] Synaptotagmin VII modulates the kinetics of dense-core vesicle exocytosis in PC12 cells. (2007)
  • Author(s): Tsuboi T, 他1名
  • Journal Title: Genes to Cells 12 Pages: 511-519
  • Peer Reviewed
• [Journal Article] Role for synaptotagmin VII in fusion pore dynamics revealed by total Internal reflection fluorescence microscopy (2007)
  • Author(s): Tsuboi T, 他1名
  • Journal Title: Neuroscience Research 58(suppl.1)
• [Journal Article] Live cell imagingによる細胞活動の可視化解析 (2007)
  • Author(s): 坪井貴司, 他1名
  • Journal Title: 日本比較内分泌学会ニュース 124 Pages: 45-53
  • NAID: 10018293579
• [Journal Article] Sustained exposure to high glucose concentrations modifies glucose signaling and the mechanics of secretory vesicle fusion in primary rat pancreatic beta-cells (2006)
  • Author(s): Tsuboi T, Ravier MA, Parton LE, Rutter GA
  • Journal Title: Diabetes 55 Pages: 1057-1065
  • Peer Reviewed
• [Journal Article] The Slp4-a linker domain controls exocytosis through interaction with Munc18-1-syntaxin-1a complex (2006)
  • Author(s): Tsuboi T, Fukuda M
  • Journal Title: Molecular Biology of the Cell 17 Pages: 2101-2112
  • NAID: 120002338802
  • Peer Reviewed
• [Journal Article] Rab3A and Rab27A cooperatively regulate the docking step of dense-core vesicle exocytosis in PC12 cells (2006)
  • Author(s): Tsuboi T, Fukuda M
  • Journal Title: Journal of Cell Science 119 Pages: 2196-2203
  • Peer Reviewed
• [Journal Article] Insulin secretion in health and disease: genomics, proteomics and single vesicle dynamics (2006)
  • Author(s): Rutter GA, Varadi A, Tsuboi T, Parton LE, Ravier MA
  • Journal Title: Biochemical Society Transactions 34 Pages: 247-250
  • Peer Reviewed
• [Journal Article] Ca^<2+> microdomains and the control of insulin secretion (2006)
  • Author(s): Rutter GA, Tsuboi T, Ravier MA
  • Journal Title: Cell Calcium 40 Pages: 539-551
  • Peer Reviewed
• [Journal Article] Rab3A and Rab27A cooperatively regulate the docking of dense-core vesicle (2006)
  • Author(s): Tsuboi T, Fukuda M
  • Journal Title: Neuroscience Research 55 (suppl. 1)
  • Peer Reviewed
• [Journal Article] Sustained exposure to high glucose concentrations modifies glucose signaling and the mechanics of secretory vesicle fusion in primary rat pancreatic beta cells (2006)
  • Author(s): Tsuboi T et al.
  • Journal Title: Diabetes 55 Pages: 1057-1065
• [Journal Article] The Slp4-a linker domain controls exocytosis through interaction with Munc18-1-syntaxin-1a complex. (2006)
  • Author(s): Tsuboi T et al.
  • Journal Title: Molecular Biology of the Cell 17 Pages: 2101-2112
  • NAID: 120002338802
• [Journal Article] Rab3A and Rab27A cooperatively regulate the docking step of dense-core vesicle exocytosis in PC12 cells. (2006)
  • Author(s): Tsuboi T et al.
  • Journal Title: Journal of Cell Science 119 Pages: 2196-2203
• [Journal Article] Ca^<2+> microdomains and the control of insulin secretion. (2006)
  • Author(s): Rutter GA et al.
  • Journal Title: Cell Calcium 40 Pages: 539-551
• [Journal Article] Insulin secretion in health and disease : genomics, proteomics and single vesicle dynamics. (2006)
  • Author(s): Rutter GA et al.
  • Journal Title: Biochemical Society Transactions 34 Pages: 247-250
• [Presentation] 細胞表面可視化法によるホルモン分泌メカニズムの解明 (2008)
  • Author(s): 坪井貴司
  • Organizer: 東京大学ナノバイオ・インテグレーション研究拠点成果発表会
  • Place of Presentation: 東京大学(東京)
  • Year and Date: 2008-12-12
• [Presentation] 細胞表面可視化法によるホルモン分泌メカニズムの解明 (2008)
  • Author(s): 坪井貴司
  • Organizer: 東京大学ナノバイオ・インテグレーション研究拠点成果発表会
  • Place of Presentation: 東京
  • Year and Date: 2008-12-12
• [Presentation] 全反射蛍光顕微鏡を用いた開口放出反応の可視化解析 (2008)
  • Author(s): 坪井貴司
  • Organizer: 「視る生物学3 -イメージングの挑戦-」文部科学省特定領域研究「植物メリステム」班会議
  • Place of Presentation: 奈良先端科学技術大学院大学(奈良)
  • Year and Date: 2008-11-21
• [Presentation] 全反射蛍光顕微鏡を用いた開口放出反応の可視化解析 (2008)
  • Author(s): 坪井貴司
  • Organizer: 文部科学省特定領域研究「植物メリステム」代表者町田泰則、NAIST植物科学研究教育推進事業
  • Place of Presentation: 奈良
  • Year and Date: 2008-11-21
• [Presentation] Rab エフェクタータンパク質によるホルモン分泌制御機構の可視化解析 (2008)
  • Author(s): 坪井貴司
  • Organizer: 第81回日本内分泌学会学術総会
  • Place of Presentation: 青森
  • Year and Date: 2008-05-17
• [Presentation] Rabエフェクタータンパク質によるホルモン分泌制御機構の可視化解析 (2008)
  • Author(s): 坪井貴司
  • Organizer: 日本内分泌学会学術総会
  • Place of Presentation: 青森
  • Year and Date: 2008-05-17
• [Presentation] Rabphilin によるホルモン分泌顆粒ドッキング制御機構の可視化解析 (2008)
  • Author(s): 坪井貴司
  • Organizer: 第85回日本生理学会大会
  • Place of Presentation: 東京
  • Year and Date: 2008-03-26
• [Presentation] Rabphilinによるホルモン分泌顆粒ドッキング制御機構の可視化解析 (2008)
  • Author(s): 坪井貴司
  • Organizer: 日本生理学会
  • Place of Presentation: 東京
  • Year and Date: 2008-03-26
• [Presentation] カテコールアミン産生腫瘍細胞を用いたホルモン分泌制御機構の可視化解析 (2007)
  • Author(s): 坪井貴司
  • Organizer: 第48回日本組織細胞化学会総会、第39回日本臨床分子形態学会総会
  • Place of Presentation: 甲府
  • Year and Date: 2007-09-28
• [Presentation] カテコールアミン産生腫瘍細胞を用いたホルモン分泌制御機構の可視化解析 (2007)
  • Author(s): 坪井貴司
  • Organizer: 日本組織細胞化学会総会、日本臨床分子形態学会総会
  • Place of Presentation: 甲府
  • Year and Date: 2007-09-28
• [Presentation] シナプトタグミン7による調節性分泌制御機構の可視化解析 (2007)
  • Author(s): 坪井貴司, 福田光則
  • Organizer: 第30回日本神経科学学会大会
  • Place of Presentation: 横浜
  • Year and Date: 2007-09-11
• [Presentation] シナプトタグミン7による調節性分泌制御機構の可視化解析 (2007)
  • Author(s): 坪井貴司 他1名
  • Organizer: 日本神経科学学会
  • Place of Presentation: 横浜
  • Year and Date: 2007-09-11
• [Presentation] Live cell imagingによるホルモン分泌顆粒ドッキング機構の解析 (2007)
  • Author(s): 坪井貴司
  • Organizer: 第3回先進治療学セミナー 順天堂大学医学部先進糖尿病治療学・内科代謝内分泌学セミナー
  • Place of Presentation: 順天堂大学医学部(東京)
  • Year and Date: 2007-06-18
• [Presentation] Live cell imagingによるホルモン分泌顆粒ドッキング機構の解析 (2007)
  • Author(s): 坪井貴司
  • Organizer: 先進治療学セミナー(順天堂大学医学部先進糖尿病治療学)
  • Place of Presentation: 東京
  • Year and Date: 2007-06-18
• [Presentation] Live cell imagingによるホルモン分泌顆粒輸送分子機構の解析 (2007)
  • Author(s): 坪井貴司
  • Organizer: 東京大学大学院医学系研究科疾患生命工学センター若手研究者セミナー
  • Place of Presentation: 東京大学(東京)
  • Year and Date: 2007-06-12
• [Presentation] Live cell imagingによるホルモン分泌顆粒輸送分子機構の解析 (2007)
  • Author(s): 坪井貴司
  • Organizer: 東京大学大学院医学系研究か疾患生命工学センター若手研究者セミナー
  • Place of Presentation: 東京
  • Year and Date: 2007-06-12
• [Presentation] Recruitment, docking, and fusion of hormone vesicles distinctively revealed by total internal reflection fluorescence microscopy in live neuroendocrine cells (2007)
  • Author(s): 坪井貴司
  • Organizer: 第2回理化学研究所フロンティア研究システム研究成果発表会
  • Place of Presentation: 理化学研究所(和光)
  • Year and Date: 2007-05-18
• [Presentation] Recruitment, docking, and fusion of hormone vesicles distinctively revealed by total internal reflection fluorescence microscopy in live neuroendocrine cells (2007)
  • Author(s): 坪井貴司
  • Organizer: 理化学研究所フロンティア研究システム研究成果発表会
  • Place of Presentation: 和光
  • Year and Date: 2007-05-18
• [Presentation] ホルモン分泌顆粒ドッキング機構の分子メカニズムの解明 (2007)
  • Author(s): 坪井貴司
  • Organizer: 第87回日本生理学会大会
  • Place of Presentation: 大阪
  • Year and Date: 2007-03-20
• [Presentation] ホルモン分泌顆粒ドッキング機構の分子メカニズムの解明 (2007)
  • Author(s): 坪井貴司
  • Organizer: 日本生理学会
  • Place of Presentation: 大阪
  • Year and Date: 2007-03-20
• [Presentation] ホルモン顆粒輸送分子機構の生細胞イメージング解析 (2006)
  • Author(s): 坪井貴司, 福田光則
  • Organizer: 日本下垂体研究会 第21回学術集会
  • Place of Presentation: 静岡
  • Year and Date: 2006-08-02
• [Presentation] Rab3AとRab27Aによるホルモン分泌顆粒ドッキング制御機構の可視化解析 (2006)
  • Author(s): 坪井貴司, 福田光則
  • Organizer: 第29回日本神経科学会大会
  • Place of Presentation: 京都
  • Year and Date: 2006-07-21
• [Presentation] Role of Slp family proteins in regulated exocytosis in neuroendocrine cells (2006)
  • Author(s): Tsuboi T, Fukuda M
  • Organizer: 20th IUBMB International Congress of Biochemistry and Molecular Biology and 11th FAOBMB Congress
  • Place of Presentation: 京都
  • Year and Date: 2006-06-20
• [Remarks]
  • URL: http://bio.c.u-tokyo.ac.jp/labs/tsuboi/
• [Remarks]
  • URL: http://bio.c.u-tokyo.ac.jp/labsitsuboil
• [Remarks]
  • URL: http://bio.c.u-tokyo.ac.jp/labs/tsuboi/