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Estimation of serine protease mRNA in glial cells after ischemic and hypoxic conditions.

Research Project

Project/Area Number 09671416
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cerebral neurosurgery
Research InstitutionASAHI UNIVERSITY (1998)
Gifu University (1997)

Principal Investigator

ANDOH Takashi  ASAHI UNIVERSITY,NEUROSURGERY,PROFESSOR, 歯学部, 教授 (90126722)

Co-Investigator(Kenkyū-buntansha) DEGUCHI Kazuki  GIFU UNIVERSITY,NEUROSUREGRY,ASSISTANT PROFESSOR, 医学部, 講師 (60217445)
TAKENAKA Katsunobu  GIFU UNIVERSITY,NEUROSUREGRY,RESEARCH ASSOCIATE, 医学部, 助手 (00283292)
YAMADA Mikito  ASAHI UNIVERSITY,NEUROSURGERY,RESEARCH ASSOCIATE, 歯学部, 助手 (40308672)
KUBOTA Yoshinori  ASAHI UNIVERSITY,NEUROSURGERY,ASSISTANT PROFESSOR, 歯学部, 講師 (60278207)
山川 弘保  岐阜大学, 医学部, 助手 (80291392)
中島 茂  岐阜大学, 医学部, 助教授 (60188935)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1998: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1997: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsSerin protease / tissue plasminogen activator / r-gsp / mRNA / ischemia / hypoxia / PC12 cell / apoptosis / 虚血 / プロテアーゼ / PLD / 分化誘導 / グリア / C6 / 海馬 / mRNA
Research Abstract

It is well-known that Caspase, kinds of Cistein proteases, are concerned in neuronal cell death. On the other hand, the roles of tPA, uPA and r-gsp, kinds of Serin proteases, for neuronal cells have not yet been fully clarified. We investigated changes in tPA and uPA mRNA expressions in the rat brain after middle cerebral artery (MCA) occlusion and PC12 cells under hypoxic condition.
Focal cerebral ischemia was produced by MCA occlusion using an intraluminal suture inserted from right internal carotid artery in male Sprague-Dawley rats (280 to320 g) under anesthesia with a mixture of fluothane. After rats were decapitated at 6 hours of MCA occlusion, total RNA were extracted from each hippocampus of bilateral hemispheres.
PC12 cell line was maintained in medium supplemented with 10% FBS, 5% horse serum in a humidified atmosphere containing 5% C02 at 37゚C.After 24h of incubation in the medium containing 2% FBS, cells were placed into hypoxic condition (multigas incubator ; set to 1.0% 02, with 5% C02 and the balance N2). Total RNA were extracted after exposure of hypoxia for 0, 3, 6, 12 and 24 hours. Extracted RNAs were analysed by RT-PCR technique using specific primer of tPA, uPA and r-gsp for investigation of changes in these mRNA expressions in each period.
Up-regulation of tPA and uPA mRNA were revealed in the rat hippocampus following 6 hours ischemia. The level of tPA mRNA was increased from the period of 6 hours exposure of hypoxia in PC12 cell, rustled in apoptoic changes. There is a tendency of up-regulation of r-gsp expression in ischemic condition. However, the level of uPA mRNA was not revealed significant changes. These results suggest that some of Serine proteases, especially tPA or r-gsp may play a role in the neuronal damage after ischemia or hypoxia.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] T Itoh, et al: "The effect of tissue plasminogen activator(t-PA)on proximal middle cerebral artery 8MCA) thromnosis in hamster." Eurpean J Neurology. 5(suppl). s51- (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S Yoshimura, et al: "Ceramide formation leads to caspase-3 activation during hypoxic PC12 cell death." J Biol Chem. 273. 6921-6927 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 林 克彦他: "頚部内頚動脈狭窄におけるself-expandable stentの使用経験" 映像情報. 30. 1355-1357 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T Itoh, S-I Yoshimura, A Okumura, K Takenaka, H Yamakawa, , T Andoh, N Sakai: "The effect of tissue plasminogen activator (t-PA) on proximal middle cerebral artery (MCA) thrombosis in hamster." European Journal of Neurology. 5(Suppl1). S49 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] S Yoshimura, Y Banno, S Nakashima, K Takenaka, H Sakai, Y Nishimura, N Sakai, S Shimizu, Y Eguchi, Y Tsujimoto, Y Nozawa: "Ceramide formation leads to caspase-3 activation during hypoxic PC12 cell death." J Biol Chem. 273. 6921-6927 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Katunobu Takenaka, Takeshi Itoh, Shin-ichi Yoshimura, Katsuhiko Hayashi, Noboru Sakai: "Up-regulation of tissue and urolonase plasminogen activator (tPA and uPA) mRNA following hypoxic conditions." Proceeding of AANS Annual Meeting. (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] T Itoh,et al: "The effect of tissue plasminogen activator (t-PA)on proximal middle cerebral artery 8MCA)thromnosis in hamster." Eurpean J Neurology. 5(suppl). s51 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S Yoshimura,et al: "Ceramide formation leads to caspase-3 activation during hypoxic PC12 cell death." J Biol Chem. 273. 6921-6927 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 林 克彦他: "頚部内頚動脈狭窄におけるself-expandable stentの使用経験" 映像情報. 30. 1355-1357 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] S Yoshimura, et al: "Changes in activiting and mRNA levels of phospholipase D(PLD) during ceramide-induced apoptosis in rat C6 glial cells." J Neurochemistry. 69. 713-720 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K Takenaka, et al: "Detection of transferrin in CSF from SAH patients,which induces nitric oxide synthase mRNA in cultured smooth muscle cells." J Cerebral Blood Flow and Metabolism. 17 Suppl. S481 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 坂井 昇他: "グリオーマ細胞の分化誘導の試み" 脳外. 25. 875-882 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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