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Imaging of quantitative changes in surface expression of ion channels using isolated perfused hearts

Research Project

Project/Area Number 26670118
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field General pharmacology
Research InstitutionYamagata University

Principal Investigator

ISHII KUNIAKI  山形大学, 医学部, 教授 (10184459)

Co-Investigator(Kenkyū-buntansha) 永澤 悦伸  東邦大学, 薬学部, 講師 (40513057)
Project Period (FY) 2014-04-01 – 2017-03-31
Project Status Completed (Fiscal Year 2016)
Budget Amount *help
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2014: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
KeywordsKチャネル / イメージング / 心臓 / GFP変異体 / pH / チャネル / pH感受性 / 心筋 / 受容体
Outline of Final Research Achievements

KCNQ1 is the pore-forming subunit of the slowly activating delayed rectifier K+ channel that plays an important role in repolarization of cardiac action potential. This study was designed to develop the imaging method to visualize changes of surface expression of KCNQ1 in the isolated perfused animal hearts.
1.pHluorin (pHl), the GFP mutant with higher pH sensitivity, was fused to the extracellular region of KCNQ1 (KCNQ1-pHl). α1 adrenergic receptor (α1AR) was co-expressed with KCNQ1-pHl in HEK293 cells, and α1AR was stimulated to cause endocytosis of KCNQ1-pHl. Since inside of early endosome is acidic, pHl fluorescence was expected to be diminished. However, activation of α1AR did not lead to significant changes in pHl fluorescence.
2.To obtain a GFP mutant that has pH sensitivity higher than pHl, we started screening of cDNA clones using the E. coli protein expression system. Unfortunately, we could not obtain such a GFP mutant so far.

Report

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

    (22 results)

All 2017 2016 2015 2014 2013

All Journal Article (6 results) (of which Int'l Joint Research: 1 results,  Peer Reviewed: 6 results,  Open Access: 3 results,  Acknowledgement Compliant: 2 results) Presentation (16 results)

  • [Journal Article] Licochalcones extracted from Glycyrrhiza inflata inhibit platelet aggregation accompanied by inhibition of COX-1 activity2017

    • Author(s)
      Okuda-Tanino A, Sugawara D, Tashiro T, Iwashita M, Obara Y, Moriya T, Tsushima C, Saigusa D, Tomioka Y, Ishii K, Nakahata N
    • Journal Title

      PLoS One

      Volume: 12 Issue: 3 Pages: e0173628-e0173628

    • DOI

      10.1371/journal.pone.0173628

    • Related Report
      2016 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] ERK5 induces ankrd1 for catecholamine biosynthesis and homeostasis in adrenal medullary cells2016

    • Author(s)
      小原 祐太郎、長澤 隆介、根本 亙、Michael J. Pellegrino、高橋 麻穂、Beth A. Habecker、Philip J.S. Stork、一柳 統、伊藤 裕美、冨田 善彦、石井 邦明、中畑 則道
    • Journal Title

      Cellular Signalling

      Volume: 28 Issue: 3 Pages: 177-189

    • DOI

      10.1016/j.cellsig.2015.12.016

    • Related Report
      2015 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Phosphorylation of ERK5 on Thr732 is associated with ERK5 nuclear localization and ERK5-dependent transcription.2015

    • Author(s)
      本多 拓人、小原 祐太郎、山内 新、 Anthony D. Couvillon、 Justin J. Mason、石井 邦明、中畑 則道
    • Journal Title

      PLoS ONE

      Volume: 10 Issue: 2 Pages: e0117914-e0117914

    • DOI

      10.1371/journal.pone.0117914

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access / Acknowledgement Compliant
  • [Journal Article] Imaging intraorganellar Ca2+ at subcellular resolution using CEPIA.2014

    • Author(s)
      Suzuki, J., Kanemaru, K., Ishii, K., Ohkura, M., Okubo, Y., and Iino, M.
    • Journal Title

      Nat. Commun.

      Volume: 5 Issue: 1 Pages: 4153-4153

    • DOI

      10.1038/ncomms5153

    • Related Report
      2014 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Is PIP2 involved in the insulin effect?2014

    • Author(s)
      石井 邦明、Minghua Wu、小原 祐太郎
    • Journal Title

      Channels

      Volume: 8 Issue: 5 Pages: 391-392

    • DOI

      10.4161/19336950.2014.948758

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Journal Article] Insulin suppresses IKs (KCNQ1/KCNE1) currents, which require β-subunit KCNE1.2013

    • Author(s)
      呉 明華、小原 祐太郎、野呂田 郁夫、永澤 悦伸、石井 邦明
    • Journal Title

      Pflugers Archiv - Eur J Physiol

      Volume: - Issue: 5 Pages: 937-946

    • DOI

      10.1007/s00424-013-1352-7

    • Related Report
      2014 Research-status Report
    • Peer Reviewed
  • [Presentation] Roles of ERK5 in ion channel expression in PC12 cells2017

    • Author(s)
      Kashino Y, Obara Y, Ishii K
    • Organizer
      第90回日本薬理学会年会
    • Place of Presentation
      長崎ブリックホール(長崎県長崎市)
    • Year and Date
      2017-03-16
    • Related Report
      2016 Annual Research Report
  • [Presentation] ラット肺静脈心筋細胞における過分極活性型Cl-チャネルの分子生物学的研究2016

    • Author(s)
      岡本洋介,石井邦明
    • Organizer
      第26回日本循環薬理学会
    • Place of Presentation
      信州大学(長野県松本市)
    • Year and Date
      2016-12-02
    • Related Report
      2016 Annual Research Report
  • [Presentation] 肺動脈血管平滑筋細胞に発現する内因性のCav1.2はアゼルニジピンにより量的修飾を受ける2016

    • Author(s)
      那須史明,倉上和也,小原祐太郎,石井邦明
    • Organizer
      第67回日本薬理学会北部会
    • Place of Presentation
      北海道大学(北海道札幌市)
    • Year and Date
      2016-09-30
    • Related Report
      2016 Annual Research Report
  • [Presentation] ラット肺静脈心筋細胞における過分極活性型Cl-チャネルの薬理学的研究2016

    • Author(s)
      岡本洋介,石井邦明
    • Organizer
      第67回日本薬理学会北部会
    • Place of Presentation
      北海道大学(北海道札幌市)
    • Year and Date
      2016-09-30
    • Related Report
      2016 Annual Research Report
  • [Presentation] Intra-organellar Ca2+ signaling imaged using genetically encoded Ca2+ indicators2016

    • Author(s)
      Suzuki J, Kanamaru K, Ishii K, Ohkura M, Ohkubo Y, Iino M
    • Organizer
      第89回日本薬理学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2016-03-11
    • Related Report
      2015 Research-status Report
  • [Presentation] Effects of high glucose on Kv1.5 and Kir2.1 channels2016

    • Author(s)
      Jimbu R, Nagasawa Y, Ishii K
    • Organizer
      第89回日本薬理学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2016-03-10
    • Related Report
      2015 Research-status Report
  • [Presentation] Impact of hyper polarization-activated Cl- current on arrhythmogenecity of rat pulmonary vein cardiomyocytes2016

    • Author(s)
      Okamoto Y, Ono K, Takano M, Ishii K
    • Organizer
      第89回日本薬理学会年会
    • Place of Presentation
      パシフィコ横浜(神奈川県横浜市)
    • Year and Date
      2016-03-09
    • Related Report
      2015 Research-status Report
  • [Presentation] Cav1.2はアゼルニジピンにより量的修飾をうける.2015

    • Author(s)
      那須史明,倉上和也,小原祐太郎,石井邦明
    • Organizer
      第66回日本薬理学会北部会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2015-09-18
    • Related Report
      2015 Research-status Report
  • [Presentation] 神経細胞におけるMidnolinの生理的•病理的な役割について.2015

    • Author(s)
      今井 亨,小原祐太郎,佐藤秀則,加藤丈夫,石井邦明
    • Organizer
      第66回日本薬理学会北部会
    • Place of Presentation
      富山国際会議場(富山県富山市)
    • Year and Date
      2015-09-18
    • Related Report
      2015 Research-status Report
  • [Presentation] Azelnidipine reduces cell-surface expression of Cav1.2 channel.2015

    • Author(s)
      Nasu F, Kurakami K, Ishii K
    • Organizer
      第30回日本不整脈学会学術大会・第32回日本心電学会学術集会
    • Place of Presentation
      国立京都国際会館(京都府京都市)
    • Year and Date
      2015-07-29
    • Related Report
      2015 Research-status Report
  • [Presentation] アゼルニジピンはCav1.2チャネルの細胞膜発現量を減少させる2015

    • Author(s)
      那須史明、倉上和也、石井邦明
    • Organizer
      第88回日本薬理学会年会
    • Place of Presentation
      名古屋国際会議場(名古屋市熱田区)
    • Year and Date
      2015-03-20
    • Related Report
      2014 Research-status Report
  • [Presentation] タンパク質型Ca2+インジケーターCEPIAを用いた小胞体・ミトコンドリア内腔Ca2+動態の可視化と機能解析2015

    • Author(s)
      鈴木純二、金丸和典、石井邦明、大倉正道、大久保洋平、飯野正光
    • Organizer
      第88回日本薬理学会年会
    • Place of Presentation
      名古屋国際会議場(名古屋市熱田区)
    • Year and Date
      2015-03-18
    • Related Report
      2014 Research-status Report
  • [Presentation] アゼルニジピンがCav1.2チャネルを量的に修飾する可能性について2014

    • Author(s)
      那須史明、倉上和也、石井邦明
    • Organizer
      第24回日本循環薬理学会
    • Place of Presentation
      山形テルサ(山形市)
    • Year and Date
      2014-12-05
    • Related Report
      2014 Research-status Report
  • [Presentation] ERK5とERK1/2のクロストーク機構について2014

    • Author(s)
      小原祐太郎、本田拓人、山内 新、Couvillon Anthony、Mason Justin、石井邦明、中畑則道
    • Organizer
      第65回日本薬理学会北部会
    • Place of Presentation
      コラッセふくしま(福島市)
    • Year and Date
      2014-09-27
    • Related Report
      2014 Research-status Report
  • [Presentation] アゼルニジピンがCav1.2チャネルをインターナリゼーションする可能性について2014

    • Author(s)
      那須史明、倉上和也、石井邦明
    • Organizer
      第65回日本薬理学会北部会
    • Place of Presentation
      コラッセふくしま(福島市)
    • Year and Date
      2014-09-26
    • Related Report
      2014 Research-status Report
  • [Presentation] Internalization of KCNQ1 by activation of the α1 adrenergic receptor2014

    • Author(s)
      倉上和也、野呂田郁夫、那須史明、永澤悦伸、小原祐太郎、石井邦明
    • Organizer
      第31回日本心電学会学術集会
    • Place of Presentation
      ザ・プリンスパークタワー東京(東京都港区)
    • Year and Date
      2014-07-25
    • Related Report
      2014 Research-status Report

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

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