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in vivo imaging analysis of cGMP dynamics in the smooth muscle of circulatory system

Research Project

Project/Area Number 25840050
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeMulti-year Fund
Research Field Biophysics
Research InstitutionThe University of Tokushima

Principal Investigator

HORIKAWA Kazuki  徳島大学, ヘルスバイオサイエンス研究部, 教授 (70420247)

Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2014: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2013: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Keywordsイメージング / cGMP / 血管平滑筋
Outline of Final Research Achievements

cGMP, one of the most important second messenger, is responsible for a variety of physiological events. To better understand the regulating mechanisms of cGMP signaling, quantitative analysis of cGMP dynamics with the help of indicator has been waited. Here, we engineered a genetically encoded biosensor that reports the concentration change of cGMP. It consists cGMP sensing motif whose N- and C-terminus were tagged with GFP variants, in which cGMP dynamics was reported in the form of FRET signal changes. By optimizing FRET parameters and the property of cGMP sensing motifs, we developed a series of improved indicators whose dynamic range was increased to 5-fold including high- to low-affinity variants. Our cell-based assay revealed that these indicators nicely report spatio-temporal dymnamics of cGMP associated with the spontaneous activity in the chemotacting cells which had not been detected with previously reported indicators.

Report

(3 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report
  • Research Products

    (9 results)

All 2015 2014 2013

All Journal Article (6 results) (of which Peer Reviewed: 5 results,  Open Access: 3 results,  Acknowledgement Compliant: 1 results) Presentation (3 results) (of which Invited: 1 results)

  • [Journal Article] Identification of polyethylene glycol-resistant macrophages on stealth imaging in vitro using fluorescent organosilica nanoparticles2015

    • Author(s)
      Nakamura M, Hayashi K, Nakano M, Kanadani T, Miyamoto K, Kori T, Horikawa K.
    • Journal Title

      ACS Nano.

      Volume: 9 Issue: 2 Pages: 1058-1071

    • DOI

      10.1021/nn502319r

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Recent progress in the development of genetically encoded Ca<sup>2+ </sup>indicators2015

    • Author(s)
      Horikawa K
    • Journal Title

      The Journal of Medical Investigation

      Volume: 62 Issue: 1.2 Pages: 24-28

    • DOI

      10.2152/jmi.62.24

    • NAID

      130005129691

    • ISSN
      1343-1420, 1349-6867
    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] In vivo visualization of subtle, transient and local activity of astrocytes using an ultrasensitive Ca2+ indicator.2014

    • Author(s)
      Kanemaru K, Sekiya H, Xu M, Satoh K, Kitajima N, Yoshida K, Okubo Y, Sasaki T, Moritoh S, Hasuwa H, Mimura M, Horikawa K, Matsui K, Nagai T, Iino M, Tanaka KF
    • Journal Title

      Cell Rep

      Volume: 8(1),311 Issue: 1 Pages: 311-318

    • DOI

      10.1016/j.celrep.2014.05.056

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Genetically encoded Ca2+ indicators; expanded affinity range, color hue and compatibility with optogenetics.2014

    • Author(s)
      Nagai T, Horikawa K, Saito K, Matsuda T
    • Journal Title

      Frontiers in Neuroscience

      Volume: 7 Pages: 25-25

    • DOI

      10.3389/fnmol.2014.00090

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Fluorescent probes for bio-imaging of bone2014

    • Author(s)
      Kazuki Horikawa
    • Journal Title

      THE BONE

      Volume: Vol.28 No.2

    • Related Report
      2013 Research-status Report
  • [Journal Article] Spatiotemporal regulation of Ca2+ dynamics in live cells revealed by Ca2+ imaging.2013

    • Author(s)
      Kazuki Horikawa and Takeharu Nagai
    • Journal Title

      CLINICAL CALCIUM

      Volume: Apr;23(4) Pages: 527-533

    • Related Report
      2013 Research-status Report
    • Peer Reviewed
  • [Presentation] 生理機能を可視化するFRETプローブの開発2014

    • Author(s)
      堀川一樹
    • Organizer
      第55回 日本生化学会 中国・四国支部例会
    • Place of Presentation
      愛媛大学(愛媛県松山市)
    • Year and Date
      2014-06-06
    • Related Report
      2014 Annual Research Report
    • Invited
  • [Presentation] ライブイメージングを容易にする高機能性指示薬の開発2014

    • Author(s)
      堀川一樹
    • Organizer
      第91回日本生理学会大会
    • Place of Presentation
      鹿児島大学( 鹿児島県)
    • Related Report
      2013 Research-status Report
  • [Presentation] 2個の細胞で多細胞生物は作れるか:細胞性粘菌の多細胞化を可能にする細胞の数2013

    • Author(s)
      佐藤朗、堀川一樹
    • Organizer
      第36回日本分子生物学会
    • Place of Presentation
      神戸国際会議場( 兵庫県)
    • Related Report
      2013 Research-status Report

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Published: 2014-07-25   Modified: 2019-07-29  

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