• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Two-photon laser scanning microscopy for the investigation of cardiomyocyte death during ischemia/reperfusion.

Research Project

Project/Area Number 20590862
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Circulatory organs internal medicine
Research InstitutionKyoto University

Principal Investigator

AKAO Masaharu  Kyoto University, 展開医療部, 主任研究員 (00362509)

Project Period (FY) 2008 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2010: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2009: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2008: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Keywords分子心臓病態学 / 心筋梗塞 / ミトコンドリア / イメージング
Research Abstract

An opening of the mitochondrial permeability transition pore (MPTP), which leads to loss of mitochondrial membrane potential (ΔΨ_m), is a crucial step in the cell death during myocardial ischemia/reperfusion ; reactive oxygen species (ROS) and Ca^<2+> are considered as the two major regulators of this step. We aimed to visualize the spatio-temporal relationship between ROS, Ca^<2+> and ΔΨ_m loss at cellular level in the perfused heart subjected to ischemia/reperfusion using a real-time two-photon imaging system. During ischemia/ reperfusion, ROS were mainly generated during early phase of ischemia with variable cell-to-cell specific rates and irreversible ΔΨ_m loss occurred in an all-or-none manner depending on cellular ROS level with a clear cut-off value. Cellular Ca^<2+> level concomitantly increased during ischemia, and once ROS level reached a threshold level, Ca^<2+> level increased robustly. Ischemic preconditioning attenuated both the increase of ROS and Ca^<2+> and protected a … More gainst ΔΨ_m loss. An anti-oxidant attenuated the ROS accumulation but not the Ca^<2+> increase ; it did not protect against ΔΨ_m loss. Thus, reducing only ROS accumulation does not suffice to prevent cardiomyocyte death ; suppression of both ROS and Ca^<2+> may be required.
An opening of the MPTP is the earliest event that commits cells to death, and this process is potentially a prime target for therapeutic intervention against myocardial ischemia/reperfusion. We aimed to investigate the protective effects of RNAi-mediated gene silencing of cyclophilin D (CypD), one of the putative components of the MPTP, against myocardial ischemia/reperfusion by using the two-photon laser scanning microscopy. We created an adenovirus carrying short interfering RNA (siRNA) which inactivates CypD. To investigate the effects in vivo, we monitored the spatio-temporal changes of ΔΨ_m in perfused rat hearts subjected to ischemia/reperfusion, using the two-photon imaging. Adult rats received direct intramyocardial injections of the adenovirus. Two to three days after injection, the rat hearts were perfused in Langendorff mode, and ΔΨ_m levels of individual cells were monitored. The progressive loss of ΔΨ_m during ischemia/reperfusion was significantly suppressed in CypD-siRNA-transduced cells, compared with non-transduced cells. Furthermore, the protective effect of CypD-siRNA was dose-dependent.
Thus, this novel two-photon imaging provides deeper insights into the mechanisms of cardiomyocyte death during ischemia/reperfusion, into cardioprotective therapy that targets mitochondria.. Less

Report

(4 results)
  • 2010 Annual Research Report   Final Research Report ( PDF )
  • 2009 Annual Research Report
  • 2008 Annual Research Report
  • Research Products

    (24 results)

All 2011 2010 2009 2008 Other

All Journal Article (8 results) (of which Peer Reviewed: 6 results) Presentation (14 results) Remarks (2 results)

  • [Journal Article] Direct monitoring of mitochondrial calcium levels in cultured cardiac myocytes using a novel fluorescent indicator protein, GCaMP2-mt.2011

    • Author(s)
      Iguchi M, Kato M, Nakai J, Takeda T, Matsumoto-Ida M, Kita T, Kimura T, Akao M.
    • Journal Title

      Int J Cardiol Epub ahead of print

    • Related Report
      2010 Final Research Report
  • [Journal Article] Direct monitoring of mitochondrial calcium levels in cultured cardiac myocytes using a novel fluorescent indicator protein, GCaMP2-mt.2011

    • Author(s)
      Moritake Iguchi, et al.
    • Journal Title

      Int J Cardiol.

      Volume: (Epub ahead of print)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The targeting of cyclophilin D by RNAi as a novel cardioprotective therapy : evidence from two-photon imaging.2009

    • Author(s)
      Kato M, Akao M, Matsumoto-Ida M, Makiyama T, Iguchi M, Takeda T, Shimizu S, Kita T.
    • Journal Title

      Cardiovasc Res. 83(2)

      Pages: 335-44

    • Related Report
      2010 Final Research Report
  • [Journal Article] p300 plays a critical role in maintaining cardiac mitochondrial function and cell survival in postnatal hearts.2009

    • Author(s)
      Nakagawa Y, Kuwahara K, Takemura G, Akao M, Kato M, Arai Y, Takano M, Harada M, Murakami M, Nakanishi M, Usami S, Yasuno S, Kinoshita H, Fujiwara M, Ueshima K, Nakao K.
    • Journal Title

      Circ Res. 105(8)

      Pages: 746-54

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] The targeting of cyclophilin D by RNAi as a novel cardioprotective therapy : evidence from two-photon imaging.2009

    • Author(s)
      Masashi Kato, et al.
    • Journal Title

      Cardiovascular Research 83

      Pages: 335-344

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] The targeting of cyclophilin D by RNAi as a novel cardioprotective therapy : evidence from two-photon imaging2009

    • Author(s)
      Masashi Kato, et al
    • Journal Title

      Cardiovascular Research (in press)

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] A novel SCN5A gain-of-function mutation M1875T associated with familial atrial fibrillation.2008

    • Author(s)
      Takeru Makiyama, et al.
    • Journal Title

      Journal of the American College of Cardiology 52

      Pages: 1326-34

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Mutation analysis of the glycerol-3 phosphate dehydrogenase-1 like (GPD1L) gene in Japanese patients with Bjugada syndrome.2008

    • Author(s)
      Takeru Makiyama, et al.
    • Journal Title

      Circulation Journal 72

      Pages: 1705-6

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Presentation] Simultaneous monitoring of reactive oxygen species, calcium and mitochondrial potential during myocardial ischemia by real-time two-photon imaging of perfused rat hearts.2011

    • Author(s)
      Moritake Iguchi
    • Organizer
      American Heart Association Scientific Sessions 2010
    • Place of Presentation
      Chicago, USA.
    • Related Report
      2010 Annual Research Report
  • [Presentation] Impact of diabetes and acute hyperglycemia on myocardial ischemia/reperfusion injury and ischemic preconditioning : evidence from real-time two-photon imaging.2011

    • Author(s)
      Masashi Kato
    • Organizer
      American Heart Association Scientific Sessions 2010
    • Place of Presentation
      Chicago, USA.
    • Related Report
      2010 Annual Research Report
  • [Presentation] Exploring Mitochondria in the Intact Ischemic Heart with Real-time Two-photon Imaging : Insights into the Novel Anti-ischemia/Reperfusion Therapy.2010

    • Author(s)
      Masaharu Akao
    • Organizer
      日本循環器学会学術集会
    • Place of Presentation
      京都
    • Year and Date
      2010-03-06
    • Related Report
      2009 Annual Research Report
  • [Presentation] Exploring Mitochondria in the Intact Ischemic Heart with Real-time Two-photon Imaging: Insights into the Novel Anti-ischemia/Reperfusion Therapy.2010

    • Author(s)
      Masaharu Akao., et al.
    • Organizer
      第74回日本循環器病学会学術集会総会
    • Place of Presentation
      京都
    • Related Report
      2010 Final Research Report
  • [Presentation] Impact of Diabetes and Acute Hyperglycemia on Myocardial Ischemia/Reperfusion Injury and Ischemic Preconditioning: Evidence from Real-time Two-photon Imaging.2010

    • Author(s)
      Masashi Kato., et al.
    • Organizer
      第74回日本循環器病学会学術集会総会
    • Place of Presentation
      京都
    • Related Report
      2010 Final Research Report
  • [Presentation] Simultaneous monitoring of reactive oxygen species, calcium and mitochondrial potential during myocardial ischemia by real-time two-photon imaging of perfused rat hearts.2010

    • Author(s)
      Moritake Iguchi., et al.
    • Organizer
      American Heart Association Scientific Sessions 2010.
    • Place of Presentation
      Chicago, USA.
    • Related Report
      2010 Final Research Report
  • [Presentation] Impact of diabetes and acute hyperglycemia on myocardial ischemia/reperfusion injury and ischemic preconditioning : evidence from real-time two-photon imaging.2010

    • Author(s)
      Masashi Kato., et al.
    • Organizer
      American Heart Association Scientific Sessions 2010.
    • Place of Presentation
      Chicago, USA.
    • Related Report
      2010 Final Research Report
  • [Presentation] Simultaneous monitoring of reactive oxygen species, calcium and mitochondrial potential during myocardial ischemia by real-time two-photon imaging of perfused rat hearts.2010

    • Author(s)
      Moritake Iguchi., et al.
    • Organizer
      第75回日本循環器病学会学術集会総会
    • Place of Presentation
      横浜
    • Related Report
      2010 Final Research Report
  • [Presentation] Visualization Of Accumulating Reactive Oxygen Species During Myocardial Ischemia By Real-time Two-photon Imaging Of Perfused Rat Hearts.2009

    • Author(s)
      Moritake Iguchi
    • Organizer
      American Heart Association
    • Place of Presentation
      New Orleans, USA
    • Year and Date
      2009-11-16
    • Related Report
      2009 Annual Research Report
  • [Presentation] Direct monitoring of mitochondrial calcium levels in cultured cardiomyocytes using a novel fluorescent indicator protein, GCaMP2-mt.2009

    • Author(s)
      Moritake Iguchi
    • Organizer
      American Heart Association
    • Place of Presentation
      New Orleans, USA
    • Year and Date
      2009-11-16
    • Related Report
      2009 Annual Research Report
  • [Presentation] Visualization of Accumulating Reactive Oxygen Species during Myocardial Ischemia by Real-time Two-photon Imaging of Perfused Rat Hearts.2009

    • Author(s)
      Moritake Iguchi., et al.
    • Organizer
      第73回日本循環器病学会学術集会総会
    • Place of Presentation
      大阪
    • Related Report
      2010 Final Research Report
  • [Presentation] An Important Role of p300 on Mitochondrial Gene Expression and Function and Tolerance to Ischemia in the Heart.2009

    • Author(s)
      Yasuaki Nakagawa., et al.
    • Organizer
      第73回日本循環器病学会学術集会総会
    • Place of Presentation
      大阪
    • Related Report
      2010 Final Research Report
  • [Presentation] Visualization Of Accumulating Reactive Oxygen Species During Myocardial Ischemia By Real-time Two-photon Imaging Of Perfused Rat Hearts.2009

    • Author(s)
      Moritake Iguchi., et al.
    • Organizer
      American Heart Association Scientific Sessions 2009.
    • Place of Presentation
      Orlando, USA.
    • Related Report
      2010 Final Research Report
  • [Presentation] Direct monitoring of mitochondrial calcium levels in cultured cardiomyocytes using a novel fluorescent indicator protein, GCaMP2-mt.2009

    • Author(s)
      Moritake Iguchi., et al.
    • Organizer
      American Heart Association Scientific Sessions 2009.
    • Place of Presentation
      Orlando, USA.
    • Related Report
      2010 Final Research Report
  • [Remarks]

    • URL

      http://kyoto-u-cardio.jp/kisokenkyu/bio/

    • Related Report
      2009 Annual Research Report
  • [Remarks]

    • URL

      http://www.kuhp.kyoto-u.ac.jp/~cardiovasc_med/kenkyushitsu/kenkymshitsu_bio.html

    • Related Report
      2008 Annual Research Report

URL: 

Published: 2008-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi