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Histological study on the gene expression of several proteins related to the intracellular Ca-signals.

Research Project

Project/Area Number 04404020
Research Category

Grant-in-Aid for General Scientific Research (A)

Allocation TypeSingle-year Grants
Research Field General anatomy (including Histology/Embryology)
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

KONDO Hisatake  Tohoku University School of Medicine Anatomy, Professor, 医学部, 教授 (20004723)

Co-Investigator(Kenkyū-buntansha) GOTO Kaoru  Tohoku University School of Medicine Anatomy, Assistant Professor, 医学部, 助手 (30234975)
Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥26,000,000 (Direct Cost: ¥26,000,000)
Fiscal Year 1993: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1992: ¥22,000,000 (Direct Cost: ¥22,000,000)
KeywordsCa-signal / gene expression / rat / in situ hybridization / gene cloning / protein kinase / プロテインキナーゼ / in situ hybridization / カルシウム結合蛋白質 / プロティンキナーゼ / 免疫組織化学 / in situ hybidization
Research Abstract

The gene cloning, sequencing and expression localization of several proteins initimately related to the intracellular Ca-signal mechanism were undertaken in the present study. These proteins included calbindin calreticulin, Ca/calmodulin dependent protein kinase(CaMK) II and IV, diacylglycerol kinase(DGK), calpain, calpastatin, 14-3-3 protein (presumptive regulator for Ca-dependent protein kinase C) ; and further protein phosphatases (PP-2A, 2B, 2C), -adrenergic receptor kinase and neuronal and non-neuronal enolases. By in situ hybridization histochemistry using specific cDNA probes for each protein which were obtained by PCR amplification, spatial and temporal heterogeneity in the expression intensity was remarkable in developing and mature rat brain. For example, cerebellar granule cells expressed intensely CaMKII and IV and PP subunits and subtypes, while no significant expression for calbindin and calreticulin was detected in these cells. This heterogeneity suggests the functional heterogeneity of these Ca-signal-related proteins in different neuronal populations, and further suggests the existence of multiple isoforms for some of these proteins. The latter possibility was pursued by using DGK and CaMKIV as targets. As a result, 80 kDa- and 90 kDa-DGKs were identified by the gene cloning. They showed 58% identity to each other and contained zinc finger-like sequences, E-F hand motifs and ATP-binding sites. The gene expression for 80 kDa-DGK was confined to oligodendrocytes, suggesting the involvement of this isoform in Ca-signals related to the myelin formation. The gene of 90 kDa-DGK was expressed predominantly in neurons of the caudate putamen, suggesting its involvement in Ca-signals related the dopaminergic transmission. In addition, a cDNA encoding CaMKIV -subtype was isolated and sequenced from rat brain. Although addition of 28 amino acids in its amino terminal region was the only difference of the -subtype from the *, the diffirential gene expression fo

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (10 results)

All Other

All Publications (10 results)

  • [Publications] 後藤薫,近藤尚武: "Molecular cloning and expression of a 90-kDa diacylglycerol kinase that predominantly localizes in neurons" Proceedings of National Academy of Sciences USA. 90. 7598-7602 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 阪上洋行,近藤尚武: "Differential expression of mRNAs encoding γ and δ subunits of Ca^<2+>/calmodulin-dependent protein kinase typeII(CaM kinaseII)in the mature and postnatally developing rat brain" Molecular Brain Research. 20. 51-63 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 阪上洋行,近藤尚武: "Cloning and sequencing of a gene encoding the β polypeptide of Ca^<2+>/calmodulin-dependent protein kinaseIV and its expression confined to the mature cerebellar granule cells" Molecular Brain Research. 19. 215-218 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 後藤薫,岩本竜明,近藤尚武: "Localization of mRNAs for calpain and calpastatin in the adult rat brain by in situ hybridization histochemistry" Molecular Brain Research. 印刷中. (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 渡辺雅彦,他4名そして近藤尚武: "Developmental regulation of neuronal expression for the η subtype of the 14-3-3 protein,a putative regulatory protein for protein kinase C" Developmental Brain Research. 73. 225-235 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 渡辺雅彦 他4名そして近藤尚武: "Molecular cloning of rat cDNAs for β and γ subtypes of 14-3-3 protein and developmental changes in expression of their mRNAs in the nervous system" Molecular Brain Research. 17. 135-146 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] K.Goto,M.WATACABE H.KONDO,H.TUASA F.SAKANE,H.KANOH: "Gene dloning sequence expession and in situ localization of 80KDa diactlglycerol Kinade specific to oligodendrocgte of rat brain" Molecrlar Brain Research. 16. 75-87 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.SAKAMI,M.WATANABE H.KONDO: "Gene expression of Ca2+/calmodilin-dependent protein Kinase of the cerebellar granule cell typr or type IV in the mature and deveoping rat brain" Molecular Brain Research. 16. 20-28 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] M.WATANBE,T.ISOBE T.Ichimura,R.KUWANO Y.TAKAHASHI,H.KONDO: "Molecular cloning of pat cDNAs for B and subtypes of 14-3-3 protein and developmental changes in expression of their mRNAs in the nervous system" Molecilar Brain Resedrch. (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] M.WATANABE,T.NAGAMINE K.SAKIMURA,Y.TAKAHASHI H.KONDO: "Developmental study of the gene expression for X and B subumits of enolase in the rat brain by in situ hybridization histochemistry" Journal Comparative Newology. 326. 1-9 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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