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Analysis of structure and function of proteins involved in membrane fusion using two-photon fluorescence lifetime imaging

Research Project

Project/Area Number 26460290
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field General physiology
Research InstitutionThe University of Tokyo

Principal Investigator

Takahashi Noriko  東京大学, 大学院医学系研究科(医学部), 講師 (60332178)

Research Collaborator WATANABE Satoshi  
NOGUCHI Jun  
SAWADA Wakako  
HASAN Ucar  
KASAI Haruo  
Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000)
Fiscal Year 2016: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords神経伝達 / 蛋白質 / 蛍光 / 2光子励起 / 開口放出 / 膜融合 / 神経 / シナプス / 伝達 / インスリン / 蛍光色素 / 内分泌 / 分泌 / シナプス伝達 / 蛍光寿命 / イメージング
Outline of Final Research Achievements

Membrane fusion is involved in neurotransmitter release and hormone secretion. Using two-photon fluorescence lifetime imaging and fluorescence labeling techniques of proteins, we measured assembly of proteins involved in membrane fusion, and detected the conformational change during the exocytosis. We detected the significant assembly of SNARE molecules in the presynaptic terminals, and oligomerization with the linker of SNAP25 was required for the ultrafast exocytosis. Moreover, such assembly was considered to be related with release probability. However, in the resting conditions in pancreatic beta cells, seldom assebly was detected, indicating that stimulants those increase cytosolic calciumconcentration is neccesary for assembling SNARE molecules.

Report

(4 results)
  • 2016 Annual Research Report   Final Research Report ( PDF )
  • 2015 Research-status Report
  • 2014 Research-status Report
  • Research Products

    (17 results)

All 2017 2016 2015 2014 Other

All Journal Article (8 results) (of which Int'l Joint Research: 3 results,  Peer Reviewed: 8 results,  Open Access: 6 results,  Acknowledgement Compliant: 3 results) Presentation (6 results) (of which Invited: 5 results) Remarks (3 results)

  • [Journal Article] Munc18b increases insulin granule fusion, restoring deficient insulin secretion in type-2 diabetes human and Goto-Kakizaki rat islets with improvement in glucose homeostasis.2017

    • Author(s)
      Qin T, Liang T, Zhu D, Kang Y, Xie L, Dolai S, Sugita S, Takahashi N, Ostenson CG, Banks K, Gaisano HY
    • Journal Title

      EBioMedicine

      Volume: 16 Pages: 262-274

    • DOI

      10.1016/j.ebiom.2017.01.030

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Design and synthesis of a new chromophore, 2-(4-nitrophenyl)benzofuran, for two-photon uncaging using near-IR light2016

    • Author(s)
      Komori, N.; Jakkampudi, S.; Motoishi, R.; Abe, M.; Kamada, K.; Furukawa, K.; Katan, C.; Sawada, W.; Takahashi, N.; Kasai, H.; Xue, B.; Kobayashi, T.
    • Journal Title

      Chemical Communications

      Volume: 52 Issue: 2 Pages: 331-334

    • DOI

      10.1039/c5cc07664a

    • Related Report
      2016 Annual Research Report 2015 Research-status Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Design and Synthesis of a 4-Nitrobromobenzene Derivative Bearing an Ethylene Glycol Tetraacetic Acid Unit for a New Generation of Caged Calcium Compounds with Two-Photon Absorption Properties in the Near-IR Region and Their Application in Vivo2016

    • Author(s)
      Satish Jakkampudi, Manabu Abe, Naomitsu Komori Ryukichi Takagi, Ko Furukawa, Claudine Katan, Wakako Sawada, Noriko Takahashi, and Haruo Kasai
    • Journal Title

      ACS Omega

      Volume: 1 Issue: 2 Pages: 193-201

    • DOI

      10.1021/acsomega.6b00119

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Opposing roles for SNAP23 in secretion in exocrine and endocrine pancreatic cells2016

    • Author(s)
      Masataka Kunii, et al,
    • Journal Title

      J. Cell Biol.

      Volume: 215 Issue: 1 Pages: 121-138

    • DOI

      10.1083/jcb.201604030

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] State-dependent diffusion of actin-depolymerizing factor/cofilin underlies the enlargement and shrinkage of dendritic spines.2016

    • Author(s)
      Noguchi, J., Hayama, T., Watanabe, S., Ucar, H., Yagishita, S., Takahashi, N. & Kasai, H.
    • Journal Title

      Scientific Reports

      Volume: 6 Issue: 1 Pages: 32897-32897

    • DOI

      10.1038/srep32897

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Two-photon fluorescence lifetime imaging of primed SNARE complexes in presynaptic terminals and β cells.2015

    • Author(s)
      Takahashi N#, Sawada W#, Noguchi J#, Watanabe S#, Ucar H, Hayashi-Takagi A, Yagishita S, Ohno M, Tokumaru H & Kasai H. (#Equally contributed).
    • Journal Title

      Nature Communications

      Volume: 6 Issue: 1 Pages: 8531-8531

    • DOI

      10.1038/ncomms9531

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Imaging Analysis of Insulin Secretion with Two-Photon Microscopy2015

    • Author(s)
      Takahashi N
    • Journal Title

      Biological and Pharmaceutical Bulletin

      Volume: 38 Issue: 5 Pages: 656-662

    • DOI

      10.1248/bpb.b14-00880

    • NAID

      130005068104

    • ISSN
      0918-6158, 1347-5215
    • Related Report
      2015 Research-status Report
    • Peer Reviewed
  • [Journal Article] Imaging Analysis of Insulin Secretion with Two-photon Microscopy2015

    • Author(s)
      Noriko Takahashi.
    • Journal Title

      Biol. Pharm. Bull.

      Volume: 38 Pages: 1-5

    • NAID

      130005068104

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Presentation] Configuration of SNAP25 at the exocytic sites analyzed with 2-photon excitation FLIM2017

    • Author(s)
      Takahashi N, Watanabe S, Sawada W, Kasai H.
    • Organizer
      第94回日本生理学会大会
    • Place of Presentation
      アクトシティ浜松(静岡県浜松市)
    • Year and Date
      2017-03-29
    • Related Report
      2016 Annual Research Report
  • [Presentation] Two-photon fluorescence lifetime imaging of primed SNARE complexes.2016

    • Author(s)
      Takahashi N, Sawada W, Noguchi J, Hasan U, Watanabe S, Kasai H.
    • Organizer
      第39回日本神経科学学会大会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2016-07-22
    • Related Report
      2016 Annual Research Report
    • Invited
  • [Presentation] 膵内分泌腺の開口放出機構2016

    • Author(s)
      高橋倫子、澤田和可子、野口潤、河西春郎
    • Organizer
      第121回日本解剖学会総会・全国学術集会
    • Place of Presentation
      ビッグパレットふくしま(福島県郡山市)
    • Year and Date
      2016-03-28
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] SNARE configuration at presynaptic terminals analyzed with 2-photon excitation FLIM2015

    • Author(s)
      Noriko Takahashi, Wakako Sawada, Jun Noguchi, Satoshi Watanabe, Haruo Kasai
    • Organizer
      第38回日本神経科学大会
    • Place of Presentation
      神戸国際会議場(兵庫県神戸市)
    • Year and Date
      2015-07-29
    • Related Report
      2015 Research-status Report
    • Invited
  • [Presentation] 2-photon FLIM analysis of SNARE assembly in neuron and endocrine cells.2015

    • Author(s)
      Takahashi N, Sawada W, Watanabe S, Noguchi J, Ohno M, Kasai H.
    • Organizer
      第92回日本生理学会・第120回日本解剖学会総会 合同大会
    • Place of Presentation
      神戸
    • Year and Date
      2015-03-22
    • Related Report
      2014 Research-status Report
    • Invited
  • [Presentation] 2光子励起画像法による蛋白質複合化の評価2014

    • Author(s)
      高橋倫子、澤田和可子、渡辺惠、河西春郎
    • Organizer
      第34回表面科学学術講演会
    • Place of Presentation
      松江 くにびきメッセ
    • Year and Date
      2014-11-07
    • Related Report
      2014 Research-status Report
    • Invited
  • [Remarks] 広報・プレスリリース最新情報-神経伝達物質やインスリン分泌の新しい可視化法開発:分泌速度の謎を解明

    • URL

      http://www.m.u-tokyo.ac.jp/news/press.html#20151007

    • Related Report
      2015 Research-status Report
  • [Remarks] 神経伝達物質やインスリン分泌の新しい可視化法開発:分泌速度の謎を解明

    • URL

      http://www.m.u-tokyo.ac.jp/news/admin/release_20151006.pdf

    • Related Report
      2015 Research-status Report
  • [Remarks] http://www.bm2.m.u-tokyo.ac.jp/

    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2018-03-22  

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