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The Investigation of the protective effect of hoat shock protein on neuronal death

Research Project

Project/Area Number 12671339
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cerebral neurosurgery
Research InstitutionAkita University

Principal Investigator

KINOUCHI Hiroyuki  Akita University, School of Medicine, Lecturer, 医学部, 講師 (30241623)

Co-Investigator(Kenkyū-buntansha) ITOH Hideaki  Akita University, School of Medicine, Associate Professor, 医学部, 助教授 (80168369)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2001: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2000: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsPC12 / electroporation / heat shock protein / cerehral ischemia / fransfection / DNA / HSP / apoptosis / heat shock protein / HSP70 / transfection / 熱ショック蛋白 / hsp / トランスフェクション / 遺伝子組み換え
Research Abstract

Gene transfer into rat brain: Rats (4, 8, or 12 weeks old) were anesthetized and fixed onto a stereotaxic operating table. A burr hole was opened and 20μg of plasmid DNA (pBK-CMV-HSP70) was injected into the hippocampus or striatum using a cannula. The injection time was changed from 10 sec to 10 min. Subsequently, a pair of stainless steel electrodes was inserted the region. Electric pulses were generated with a Square Electro Porator (CUY21) and were administered at 10 Hz. Electric pulses were 2ms in duration at 50, 100, or 150 V. However, at any condition, the sufficient gene transfer was not obtained. Only a couple of neurons showed β-galactosidase positive coincidentally.
Transfection of HSP70 gene into PC12 cells: Human HSP75 cDNA was transfected using the pBK-CMV vector. Production of HSP70 protein was confirmed by Western blot analysis. The PC12 cells overexpressing HSP70 were exposed to hydrogen peroxide (50, 100, 150μM). MTT assay after 24h of exposure revealed the significant viability of HSP70-transfected PC12. The protective effect of HSP70 against oxidative injury was revealed.

Report

(3 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Okubo Atsuya: "Simultaneous induction of mitochordrial heat shock protein mRNAs in cat forebrain ischemia"Mdecular Brain Research. 84. 127-134 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Izaki Kenji: "Induction of mitochordrial heat schock protein 60 and 10 mRNAs following fransient focal cerehral ishemia in the rat"Molecular Brain Research. 88. 14-25 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ckubo A, Kinouchi H, Owada Y, Kunizuka H, Itoh H, Izaki K, Kondo H, Yoshimoto T, Mizoi K: "Simultaneous induction of mitochondrial heat shock protein mRNAs in rat forebrain ischemia"Molecular Brain Research. 84. 127-134 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Izaki K, Kinouchi H, Watanabe K, Owada Y, Okubo A, Itoh H, Kondo H, Tashima Y, Tamura S, Yoshimoto T, Mizoi K: "Induction of mitochondrial hear shock protein 60 and 10 mRNAs follorving transient focal cerebral ischemia in the rat"Molecular Brain Research. 88. 14-25 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Izaki Kenji: "Induction of mitochondrial heat shock protein 60 and 10 mRNAs following transient focal Cerebral ischemia in the rat"Molecular Brain Research. 88・1-2. 14-25 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Atsuya Okubo: "Simultaneous induction of mitochondrial heat shock protein mRNAs in rat forebrain ischemia"Molecular Brain Research. 84・1-2. 127-134 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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