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Development of new inverse-type calcium probe for analysis of inhibitory neuronal activities

Research Project

Project/Area Number 26560461
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Basic / Social brain science
Research InstitutionKyushu University

Principal Investigator

Kuge Sayuri  九州大学, 理学(系)研究科(研究院), 学術研究員 (50260104)

Co-Investigator(Renkei-kenkyūsha) ISHIHARA Takeshi  九州大学, 大学院理学研究院, 教授 (10249948)
TERAMOTO Takayuki  九州大学, 大学院理学研究院, 准教授 (90571836)
NAGAI Takeharu  大阪大学, 産業科学研究所 (20311350)
MATSUDA Tomoki  大阪大学, 産業科学研究所 (50419206)
Project Period (FY) 2014-04-01 – 2016-03-31
Project Status Completed (Fiscal Year 2015)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2014: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
Keywordsカルシウムプローブ / 神経活動 / 線虫 / インバース
Outline of Final Research Achievements

Fluorescent Ca2+ indicators, which have been improved for the last several decades are ideal only for monitoring the activation of neurons. To understand precise functions of the neuronal network, a fluorescent Ca2+ indicator that is suitable for monitoring the neuronal inhibition would be necessary. Previously reported "inverse-pericam" has unique property; the fluorescence intensity gets 7-fold dimmer upon Ca2+ binding. After creating a large mutant library of "inverse-pericam" by error-prone PCR, a variant, IP2.0, which shows nearly 20-folds dynamic range in vitro, was found. When IP2.0 was expressed in AWC neuron of C. elegans, the increase of fluorescence of IP2.0 during the stimulation of isoamyl alcohol was observed. These results suggest that odor stimuli decreases Ca2+ concentration in AWCON neurons and so IP2.0 will be an indispensable tool for studying neuronal inhibition.

Report

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

    (9 results)

All 2015 2014

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Acknowledgement Compliant: 1 results) Presentation (8 results) (of which Int'l Joint Research: 3 results)

  • [Journal Article] The peroxin Pex14p is the key component for coordinated autophagic degradation of mammalian peroxisomes by direct binding to LC3-II2015

    • Author(s)
      Jiang, L., Hara-Kuge, S., Yamashita, S., and *Fujiki, Y.
    • Journal Title

      Genes to Cells

      Volume: 20 Issue: 1 Pages: 36-49

    • DOI

      10.1111/gtc.12198

    • Related Report
      2015 Annual Research Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] Quenching-type of fluorescent Ca2+ indicator for detecting neuronal inhibition2015

    • Author(s)
      Sayuri Kuge, Tomonobu Nishihara, Tomoki Matsuda, Hironobu Furuie, Takayuki Teramoto, Takeharu Nagai, Takeshi Ishihara
    • Organizer
      2015 International Chemical Congress of Pacific Basin Societies
    • Place of Presentation
      Honolulu, Hawaii
    • Year and Date
      2015-12-15
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 線虫C.elegansの頭部神経系全体のGCaMP6fを用いた活動解析2015

    • Author(s)
      大江紗、寺本孝行、徳永旭将、広瀬修、豊島有、久下小百合、飯野雄一、吉田亮、石原健
    • Organizer
      第38回日本分子生物学会年会、第88回日本生化学会大会合同大会
    • Place of Presentation
      神戸ポートアイランド
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
  • [Presentation] 抑制性神経活動を検出する消光型蛍光カルシウムプローブタンパク質2015

    • Author(s)
      久下小百合、西原知伸、松田知己、古家博信、寺本孝行、永井健治、石原健
    • Organizer
      第38回日本分子生物学会年会、第88回日本生化学会大会合同大会
    • Place of Presentation
      神戸ポートアイランド
    • Year and Date
      2015-12-01
    • Related Report
      2015 Annual Research Report
  • [Presentation] A quenching-type of fluorescent Ca2+ indicator for detecting neuronal inhibition2015

    • Author(s)
      Sayuri Kuge, Tomonobu Nishihara, Tomoki Matsuda, Hironobu Furuie, Takayuki Teramoto, Takeharu Nagai, Takeshi Ishihara
    • Organizer
      C.elegans 2015 20th International Meeting
    • Place of Presentation
      University of California
    • Year and Date
      2015-06-24
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] Ca2+ dependent negative feedback loop in AWC olfactory neurons2015

    • Author(s)
      Mamoru Usuyama, Sayuri Kuge, Takayuki Teramoto, Takeshi Ishihara, YUishi Iwasaki
    • Organizer
      C.elegans 2015 20th International Meeting
    • Place of Presentation
      University of California
    • Year and Date
      2015-06-24
    • Related Report
      2015 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 線虫、C.elegansの単一神経細胞におけるカルシウムダイナミクス2014

    • Author(s)
      久下小百合、寺本孝行、石原健
    • Organizer
      第37回日本分子生物学会年会
    • Place of Presentation
      パシフィコ横浜
    • Year and Date
      2014-11-25 – 2014-11-27
    • Related Report
      2014 Research-status Report
  • [Presentation] Tracking of Ca2+ dynamics in a whole single neuron2014

    • Author(s)
      Sayuri Kuge, Takayuki Teramoto, Takeshi Ishihara
    • Organizer
      Neuronal Development, Synaptic Function and Behavior
    • Place of Presentation
      University of Wisconsin-Madison
    • Year and Date
      2014-07-07 – 2014-07-10
    • Related Report
      2014 Research-status Report
  • [Presentation] Electrodifusion model for Ca2+ dynamics in a whole single neuron2014

    • Author(s)
      Yuishi Iwasaki, Sayuri Kuge, Takayuki Teramoto, Takeshi Ishihara
    • Organizer
      Neuronal Development, Synaptic Function and Behavior
    • Place of Presentation
      University of Wisconsin-Madison
    • Year and Date
      2014-07-07 – 2014-07-10
    • Related Report
      2014 Research-status Report

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Published: 2014-04-04   Modified: 2017-05-10  

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