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In vivo transfection of foreign genes into the neurons of rat brain as the neurografting method

Research Project

Project/Area Number 03670684
Research Category

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

Allocation TypeSingle-year Grants
Research Field Cerebral neurosurgery
Research InstitutionOkayama University

Principal Investigator

MIZUKAWA Kiminao  Okayama University Medical School, Lecturer, 医学部, 講師 (40137154)

Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1992: ¥300,000 (Direct Cost: ¥300,000)
KeywordsDNA transfection / Lipofectin / Plasmid DNA / beta-galactosidase / Enzyme histochemistry / Immunohistochemistry / 酵素組織化学 / 免疫組織化学 / ブラスミドDNA / ベ-タガラクトシラ-ゼ
Research Abstract

First of all, I investigated the several factors controlling the efficiency of DNA transfection. I tried to use plasmid DNA as a vector for the introduction of genes into mouse muscles cells using cationic lipid-mediated DNA transfection(lipofection).
By this improvement of this lipofectin-treated DNA, plasmid DNA were easily First of all, I investigated the several factors controlling the efficiency of DNA transfection. I tried to use plasmid DNA as a vector for the introduction of genes into mouse muscles cells using cationic lipid-mediated DNA transfection(lipofection).
By this improvement of this lipofectin-treated DNA, plasmid DNA were easily injected into the mouse muscles through a microsyringe, and could be incorporated and expressed by muscle cells in the vicinity of injection point Morphologically I used two kinds of plasmid DNAs, L7RH-beta-gal and pRSVL-beta-gal, in order to detect the expression of transfected DNAs in mouse muscles cells. The L7RH-beta-gal plasmid DNA contain … More s the unclear location signal from the SV40 T-antigen gene and E.coli-beta-galactosidase gene both of which are fused to the SV40 virus enhancer-promoter. Since the produced E.coli beta-galactosidase proteins can be transported into nuclei of cells when the muscle cells were transfected with the L7RH-plasmid DNA, the nuclei should be densely stained with X-gal in a round shape. On the contrary, using the pRSVL-beta-gal, several number of the X-gal-stained mouse muscles fiber bundles were observed as blue colored muscles by light microscopy. By electron microscopic investigation, the large number of fine electron dense materials were occupied in the sarcolemma of the X-gal stained muscles.
Although we have not identified any factors controlling the efficiency of lipofectin to mouse muscles cells with plasmid DNAs, improvements of this technique would provide us with a powerful tool for studying gene action in particular muscle cells in relation to morphological and functional aspects.
Next stage, I will intend to apply this procedure to the neurografting. Less

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] ASANUMA,M.et al: "Distribution of the beta-2 adrenergic receptor messenger RNA in the rat brain by in situ hybridization histochemistry:effects of chronic reserpine treatment" Neurochemical Research,. 16. 1253-1256 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] MIZUKAWA,K.et al: "Morphological investigation of iron-induced epileptic rats:with special reference to c-fos immunohistochemistry and iron staining" The Japanese Joumal of Psychiatry and Neurology. 45. 285-289 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] MIZUKAWA,K.et al: "Changes of phorbol ester binding sites in rat brain following intracerebroventricular administration of thyrotropin-releasing hormone (TRH):an in vitro macroautoradiographic investigation" Neurochemical Research,. 17. 361-365 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] OGAWA,N.et al: "Post-ischemic administration of bifemelane hydrochloride prohibits ischemia-induced depletion of the muscarinic Ml-receptor and its mRNA in the gerbil hippocampus" Brain Research,. 591. 171-175 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Haba, K., N.Ogawa, K.Mizukawa and A.Mori: "Time course of changes in lipid peroxidation, pre- and postsynaptic cholinergic indices, NMDA receptor binding and neuronal death in the gerbil hippocampus following transient ischemia" Brain Research. 540. 116-122 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ogawa, N., K.Mizukawa, K.Haba, M.Asanuma and A.Mori: "Chronic bifemelane hydrochloride administration enhances muscarinic cholinergic receptor binding in the senescent rat brain" Journal of Medicine.22(1). 17-27 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ogawa, N. K.Haba, K.Mizukawa, M.Asanuma, H.Hirata and A.Mori: "Loss of N-methyl-D-Aspartate(NMDA) receptor binding in the rat hippocampal area at the chronic stage after transient forebrain ischemia : histological and NMDA receptor binding studies" Neurochemical Research. 16. 519-524 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Hirata, H., N.Ogawa, K.Mizukawa, K.Haba, M.Asanuma, A.Mori and Z.Ota: "Chronic lisuride hydrogen maleate administration enhances muscarinic receptor binding insenescent rat brain" Journal of International Medical Research. 19. 318-325 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Asanuma, M., N.Ogawa, K.Mizukawa, K.Haba, H.Hirata, and A.Mori: "Distribution of the beta-2 adrenergic receptor messenger RNA in the rat brain by in situ hybridization histochemistry : effects of chronic reserpine treatment" Neurochemical Research. 16(12). 1253-1256 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mizukawa, K., H.Kabuto, and A.Mori: "Morphological investigation of iron-induced epileptic rats : with special reference to c-fos immunohistochemistry and iron staining" The Japanese Journal of Psychiatry and Neurology. 45(2). 285-289 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ono, K., A.Tokunaga, K.Mizukawa, K.Kurose and H.Tanaka: "Abnormal expression of embryonic N-CAM in the developing mouse cerebellum after neonatal administration of cytosine arabinoside" Developmental Brain Research. 65. 119-122 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mizukawa, K., N.Otsuka, N.Ogawa, K.Haba, and A.Mori: "Changes of phorbol ester binding sites in rat brain following intracerebroventricular administration of thyrotropin-releasing hormone(TRH) : an in vitro macroautoradiographic investigation" Neurochemical Research. 17(4). 361-365 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Ogawa, N., M.Asanuma, K.Mizukawa, H.Hirata, H.Chou and A.Mori: "Post-ischemic administration of bifemelane hydrochloride prohibits ischemia-induced depletion of the muscarinic Ml-receptor and its m RNA in the gerbil hippocampus" Brain Research. 591. 171-175 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Mizukawa, K. and A.Mori: "Alterations in benzodiazepine and gamma aminobutyric acid (GABA)ergic receptors in El mice : in vitro macroautoradiographic investigation" Neurosciences 18. Supplement 2. (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Asanuma,M.,et al.: "Distribution of the β-2 adrenergic receptor messenger RNA in the rat brain by in situ hybridization histochemistry:effects of chronic resepine treatment" Neurochemical Research. 16. 1253-1256 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mizukawa.K.,et al: "Morphological investigation of iron-induced epileptic rats:with special reference to c-fos immunohistochemistry and iron staining" The Japanese Journal of Psychiatry and Neurology. 45. 285-289 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mizukawa,K.,et al: "Changes of phorbol ester binding sites in rat brain following intracerebroventricular administration of thyrotropin-releasing hormone(TRH):an in vitro macroautoradiographic investigation" Neurochemical Research,. 17. 361-365 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Ono,K.,et al: "Abnormal expression of embryonic N-CAM in the developing mouse cerebellum after neonatal administration of cytosine arabinoside" Developmental Brain Research. 65. 119-122 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Ogawa,N.,et al: "Post-ischemic administration of bifemelane hydrochloride prohibits ischemia-induced depletion of the muscarinic Mlreceptro and its m RNA in the gerbil hippocampus," Brain Research. 591. 171-175 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Mizukawa,K.et al: "Alterations in benzodiazepine and γ aminobutyric acid (GABA)ergic receptors in Elmice:in vitro macroautotoradiographic investigation," Neurosciences 18 Supplement 2. 18. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] Ono,K.,et al.: "Abnormal expression of embryonic neuronal cell adhesion molecule(NーCAM)in the developing mouse cerebellum after neonatal administration of cytosine arabionoside" Developmental Brain Research. 65. 119-122 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] MiZUKAWA,K.,et al.: "Changes of Phorbol ester binding sites in rat brain following intracerebroventricular administration of thyrotropinーreleasing hormone(TRH):an in vitro macroautoradiographic investigation" Neurochemical Research. 17. 361-365 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] MiZUKAWA,K,et al.: "In vivo transfection of foreign genes into mouse muscles:electron and light microscopical investigation" Brain Research.

    • Related Report
      1991 Annual Research Report

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

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