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A gene trap approach to identify a new gene involved in central nervous system development

Research Project

Project/Area Number07557092
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Cerebral neurosurgery
Research InstitutionNiigata University

Principal Investigator

KUWANO Ryozo  Niigata University, Research Laboratory for Molecular genetics, Associate Professor, 遺伝子実験施設, 助教授 (20111734)

Co-Investigator(Kenkyū-buntansha) KUMANISHI Toshiro  Niigata University, Brain Research Institute, Professor, 脳研究所, 教授 (40018601)
Project Period (FY) 1995 – 1997
Project Status Completed(Fiscal Year 1997)
Budget Amount *help
¥21,400,000 (Direct Cost : ¥21,400,000)
Fiscal Year 1997 : ¥3,600,000 (Direct Cost : ¥3,600,000)
Fiscal Year 1996 : ¥6,800,000 (Direct Cost : ¥6,800,000)
Fiscal Year 1995 : ¥11,000,000 (Direct Cost : ¥11,000,000)
Keywordsgene / cell abration / Cre-lox / diphteria toxin / gene trapping / toransgenic mice / brain / immunohistochemistry / 免疫組織科学 / トランスジェニック / ネオマオシン耐性 / マウス / 神経成長円錐 / リコンビネース / cre-lox / 遺伝子発現 / アストロサイト / 神経機能
Research Abstract

In attempt to elucidate of interaction between neuron and glia involved in neuronal function we used a strategy of cell ablation using a diphtheria toxin A fragment (DT-A) gene in transgenic mice. Based on spatial-temporal specific gene expression using an appropriate promoter such as S100 or GFAP promoter in astrocyte and a novel one for neuron discovered by gene trapping, the DT-A gene is activated in the restricted cells in the central nervous system. In addition we introduced cre-lox system for regional and stage specific regulation of the target gene. expression. Cre-recombinase mediated site specific recombination occurs on the restricted DNA sequence composed of 34 bp(loxP). DT-A activity was suppressed by insertion of transcription termination signal down stream of murine molony leukemia virus promoter. Cell ablation by cre-lox has to be verified in culture system prior to create transgenic mice. We confirmed an effect of DT-A in the transfected cells. We produced several transgenic mouse lines of which some showed ectopic expression pattern owing to a position effect.
Gene trapping provided a novel promoter expressed in the central nervous system and heart. A new construct of the target gene containing insulator sequence for cell ablation is constructed again.

Report

(4results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • 1995 Annual Research Report

Research Products

(28results)

All Other

All Publications

  • [Publications] Shirai, M.: "A gene trap strategy fo identifying the gene expressed in the embryonic nervous system." Zool.Sci.13. 277-283 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yuasa, S.: "Neuron-glia interrelations during migration of Purkinje cells in the mouse embryonic cerebellum." Int.J.Dev.Neurosci.14. 429-438 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Abe, T.K.: "The presence of the 50-kDa subunit of dynactin complex in the nerve growth cone." Biochem.Biophys.Res.Commun.233. 295-299 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Usui, H.: "cDNA cloning and mRNA expression analysis of the human neuronatin." J.Mol.Neurosci.9. 55-60 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Fujiwara, M.: "Expressions of a calcium-binding protein (spot 35/calbindin-D28k) in mouse olfactory cells: possible relationship to neuronal differentiation." Eur.Arch.Otorhinolaryngol.254. 105-109 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Itoh, Y.: "Mapping of chlesystokinin transcription in transgenic mice brain using Esherichia coli β-galactoridase reporter gene." Devl.Growth.Different.(in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Shirai, M.et al.: "A gene trap strategy for identifying the gene expressed in the embryonic nervous system." Zool.Sci.13. 277-283 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yuasa, S.et al.: "Neuron-glia interrelations during migration of Purkinje cells in the mouse embryonic cerebellum." Int.J.Devl.Neurosci.14. 429-438 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Abe, T.K.et al.: "The presence of the 50-kDa subunit of dynactin complex in the nerve growth cone." Biochem.Biophys.Res.233. 295-299 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Usui, H.et al.: "cDNA cloning and mRNA expression analysis of the human neuronatin." J.Mol.Neurosci.9. 55-60 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Fujiwara, M.et al.: "Expressions of a calcium-binding protein (spot35/calbindin-D28k) in mouse olfactory cells : possible relationship to neuronal differentiation." Eur.Arch Otorhinolaryngol. 254. 105-109 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Itoh, Y.et al.: "Mapping of chlesystokinin transcription in transgenic mice brain using Esherichia coli beta-galactoridase reporter gene." Devl.Growth.Different.(in press.).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Shirai, M.: "A gene trap strategy for identifying the gene expressed in the embrycric nervous system." Zool. Sci.13. 277-283 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Yuasa, S.: "Neuron - glia interrelations during migration of Purkinje cells in the mouse smbryanic oerebellum." Int. J. Devl.14. 429-438 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] Abe, T. K.: "The presenae of the 50 - KDa subunit of dynactin complex in the nerve growth cone." Biochem. Biophys. Res. Commun.233. 295-299 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Usui, H.: "cDNA cloning and mRNA expression analysis of the human neuronatin." J. Mol. Neurosci.9. 55-60 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Fujiwara, M.: "Expressions of a clacium - binding protein ( spot 35 / calbindin - D28k ) in mouse olfactory cells : possible relationship to neuronal differentiation." Eur. Arch. Otorhinolaryngol.254. 105-109 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Itoh, Y.: "Mapping of chlesystokinin transcription in transgenic mioe brain using Esherichia coli β - galactoridase reporter gene." Devl. Growth. Different.(in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] M.Shirai: "A gene trap strategy for identifying the gene expression in the embryonic nervous system." Zool.Sci.13. 277-283 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.Muratake: "Structural organization and chromosomal assignment of the human 14-3-3h chain gene (YWHAH)" Genomics. 36. 63-69 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Yuasa: "Neuron-glia interrelations during migration of purkinje cells in the mouse embryonic cerebellum" Int.J.Devel.Neurosci. 14. 429-438 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] S.Kawahire: "cDNA cloning of nuclear localization signal binding protein NFP60,a rat homologue of lamin B receptor,and identification of binding sites ofhuman lamin B receptor for nuclear localization signals and chromatin" J.Biochem.121(in press). (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] T.K.Abe: "Two‐dimensional analysis and sequencing of membrane proteins highly concentrated in the nerve growth cone‐enriched fraction isolated from mouse brain." Chromatography. 16. 39-44 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] H.Kuramoto: "Location of sensory nerve cells that provide calbindin‐containing laminar nerve endings in myenteric ganglia of the rat esophagus." J.Auton.Nerv.Syst.54. 126-136 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Kobayashi: "Molecular cloning of cDNAs for immunoglobulin variable regions of monoclonal anti‐idiotypic antibody specific for sigma receptors." Journal of Neuroimmunol.57. 129-135 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Ikeda: "Functional couplings of the delta‐and the kappa‐opioid receptors with the G‐protein‐activated K+channel." Biochem.Biophys.Res.Commun.208. 302-308 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] T.Kobayashi: "Molecular cloning of a mouse G‐protein‐activated K+channel(mGIRK1)and distinct distributions of three GIRK(GIRK1,2and3)mRNAs in mouse brain." Biochem.Biophys.Res.Commun.208. 1166-1173 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] M.Shirai: "A gene trap strategy for identifying the gene expression in the embryonic nervous system." Zool.Sci.13(in press). (1996)

    • Related Report
      1995 Annual Research Report

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Published : 1995-04-01   Modified : 2016-04-21  

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