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Studies on neural plasticity using transgenic mice expressing GFP-tagged PKC

Research Project

Project/Area Number 12680754
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Neurochemistry/Neuropharmacology
Research InstitutionKobe University

Principal Investigator

SAKAI Norio  Biosignal Research Center, Kobe University, Associate Professor, バイオシグナル研究センター, 助教授 (70263407)

Co-Investigator(Kenkyū-buntansha) SAITO Naoaki  Biosignal Research Center, Kobe University, Professor, バイオシグナル研究センター, 教授 (60178499)
Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,400,000 (Direct Cost: ¥2,400,000)
KeywordsPKC / transgenio mice / phosphorylation / neural plasticity / tet-regulated / Purkinje cells / プイロテインキナーゼC / 小脳プリキンエ細胞
Research Abstract

1. Development an danalysis of tet-regulated transgenic mice expressing PKC-GFP in a brain region specific manner
1) Development and analysis of the transgenic mice expressing tetracycline transactivator (tTA) under the control of various promoters which drive to express target proteins in a brain region specific manner.
In this study, we analyze transgenic mice which express tTA under the control of neuron specific enolase (NSE) and calcium-calmodulin dependent kinase n (CamKII) promoter. The former mice expressed tTA in striatum neurons and cerebellar Purkinje cells, while the latter expressed it in anterior brain including hippocampus. In addition, we developed novel transgenic mice which express tTA under the control ofL7 and PKCγ promoter.
2) Development of transgenic mice expressing PKC-GFP under the control of tetracycline-operated (TetOp) promoter
We have developed transgenic mice which can express various subtypes of GFP-fused PKC (PKCγ, PKCε, PKCδ and PKCζ) under the control ofTetOp promoter.
3) Development of tet^egulated bi-transgenic mice.
The TetOp-PKCγ-GFP mice was crossed with the NSE-tTA or CaMKII-tTA to obtain brain region specific expression of PKCγ-GFP. PKCγ-GFP was mainly expressed in striatum GABA-containing neuron and cerebellar Purkinje cells under the control of NSE promoter. In contrast, PKCγ-GFP was expressed in olfactory bulb and anterior whole brain when it was crossed with CamKn-tTA mice. Expression ofPKC-GFP was completely abolished by the treatment with doxycycline for 4 weeks and the expression was recovered by the cessation ofdoxycycline treatment.
2. Imaging of PKC translocation in vivo state using PKC-GFP transgenic mice
Cerebellar slices were prepared from these transgenic mice. Translocation of PKC-GFP was visualized by 2-photon laser scanning microscope. In Purkinje cells, PKCγ-GFP was transiently translocated to the plasma membrane by the activation of metabotropic glutamate receptors.

Report

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

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Sumioka, K 他: "Induction of 55 kDa protein of PKN by ischemia/reperfusion model of rat retina"Invest.Ophthal.Vis.Sci.. 41. 29-35 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shirai, Y 他: "Phospholipase A2 and its products are involved in the purinergic receptor-mediated translocation of protein kinase C in CHQ-K1 cells"J.Cell Sci. 113. 1335-1343 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shirai, Y 他: "Subtype-specific translocation of diacylglycerol kinase a and g and its correlation with protein kinase C Shirai, Y"J.Biol.Chem.. 275. 24760-24766 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ohmori, S 他: "Importance of PKC targeting for the phosphorylation of its substrate, myristoylated alanine-rich C-kinase substrate (MARCKS)"J.Biol.Chem.. 275. 26449-26457 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Taketoshi Kajimoto 他: "Subtype-specific Translocation of δ subtype of protein kinase C and its activation by tyrosine phosphorylation induced by ceramide in Hela cells"Mol.Cell Biol.. 21. 1769-1783 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takehiiko Ueyama 他: "Constitutively active fragment of PKN in microglia/macrophage after middle cerebral artery occlusion in rats"J.Neurochem.. 79. 903-913 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kaori Kashiwagi 他: "Importance of C1B domain for lipid messenger-induced targeting of PKC"J.Biol.Chem.. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Sumioka, K., Shirai, Y., Sakai, N., Hashimoto, T., Tanaka, C., Yamamoto, M., Takahashi, M., Ono, Y., and Saito, N.: "Induction of 55 kDA protein of PKN by ischemia/reperfusion model of rat retina"Ophthal. Vis. Sci.. 41. 29-35 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shirai, Y., Kashiwagi, K., Sakai, N. and Saito, N.: "Phospholipase A2 and its products are involvged in the purinergic receptor-mediated translocation of protein kinase C in CHO-K1 cells"J. Cell Sci.. 113. 1335-1343 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shirai, Y., Sagawa, S., Kuriyama, M., Goto, K., Sakai, N. and Saito, N.: "Subtype-specific translocation of diacylglycerol kinase a and g and its correlation with protein kinase C Shirai, Y."J. Biol. Chem.. 275. 24760-24766 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Ohmori, S., Sakai, N., Shirai, Y., Yamamoto, H., Miyamoto, E., Shimizu, N. and Saito, N.: "Importance of PKC targeting for the phosphorylation of its substrate, myristoylated alanine-rich C-kinase substrate (MARCKS)"J. Biol. Chem.. 275. 26449-26457 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kajimoto, T., Ohmori, S., Shirai, Y.,m Sakai, N. and Saito, N.: "Subtype-specific translocation of δsubtype of protein kinase C and its activation by tyrosine phosphorylation induced by ceramide in Hela cells"Mol. Cell biol.. 21. 1769-1783 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takehiko Ueyama et al: "Constitutively active fragment of PKN in microglia/macrophage after middle cerebral arthery occlusion in rats"J. Neurochem. 79. 903-913 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Kaori Kashiwagi et al: "Importancde of C1B domain for lipid messenger-induced targeting of PKC"J. Biol. Chem. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Takehiko Ueyama: "Constitutively active fragment of PKN in microglia/macrophage after middle cerebral artery occlusion in rats"J.Neurochem.. 79. 903-913 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Taketoshi Kajimoto: "Subtype-specific translocation of the δ subtype of protein kinase C and its activation by tyrosine phosphorylation induced by ceramide in HeLa cells"Mol. Cell. Biol.. 21. 1769-1783 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kaori Kashiwagi: "Importance of C1B domain for lipid messenger-induced targeting of PKC"J. Biol. Chem.. (in press). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] Chen, A: "Regulation of GFRα-1 and GFRα-2 mRNAs in rat brain by electroconvulsive seizure"Synapse. 39. 42-50 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kajimoto,T.: "Subtype-specific translocation of δ-PKC and its activation by tyrosine phosphorylation induced by ceramide in HeLa cells."Mol.Cell.Biol.. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ohmori,S.: "Importance of PKC targeting for the phosphorylation of its substrate, myristoylated alanine-rich C-kinase substrate (MARCKS)."J.Biol.Chem.. 275. 26449-26457 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Shirai,Y.: "Subtype-specific translocation of diacylglycerol kinase α and γ and its correlation with protein kinase C."J.Biol.Chem.. 275. 24760-24766 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Chen,A.: "Regulation of GFRa-1 and GFRa-2 mRNAs in rat brain by electroconvulsiveseizure."Synapse. (印刷中). (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Shirai,Y.: "Phospholipase A2 and its products are involved in the purinergic receptormediated translocation of protein kinase C in CHO-K1 cells."J.Cell Sci.. 113. 1335-1343 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sumioka,K.: "Induction of 55 kDa PKN cleavage product by ischemia/reperfusion model in the rat retina."Invest.Opthal.Vis.Sci.,. 41. 29-35 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sakai,N.: "Involvement of the actin cytoskeleton in the regulation of serotonin transporter (SET) activity : possible mechanism underlying SET regulation by protein kinase C."Neurochem.Int.. 36. 567-579 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Thome,J.: "cAMP response element-mediated gene transcriptionis upregulated by chronic antidepressnt treatment."J.Neurosci.. 20. 4030-4036 (2000)

    • Related Report
      2000 Annual Research Report

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

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