• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

FUNCTION OF NITRIC OXIDE AS A TROPHIC FACTOR SUPPORTING DIFFERENTIATION AND MATURATION OF NEURONS ; ANALYSIS USING ULTRAMICRO TECHNIQUES

Research Project

Project/Area Number 07458206
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Neurochemistry/Neuropharmacology
Research InstitutionYOKOHAMA CITY UNIVERSITY

Principal Investigator

KATO Takahiko  YOKOHAMA CITY UNIVERSITY SCHOOL OF MEDICINE,PROFESSOR, 医学部, 教授 (80010023)

Co-Investigator(Kenkyū-buntansha) NAKAYAMA Takashi  YOKOHAMA CITY UNIVERSITY,SCHOOL OF MEDICINE,LECTURER, 医学部, 講師 (90150060)
INOUE Nobuo  YOKOHAMA CITY UNIVERSITY,SCHOOL OF MEDICINE,ASSOCIATE PROFESSOR, 医学部, 助教授 (50159985)
Project Period (FY) 1995 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsNITRIC OXIDE / NITRIC OXIDE SYNTHASE (NOS) / NOS-ISOFORMS / NOS-ISOFORM mRNAs / MICRO-RT-PCR / CEREBELLAR GRANULAR LAYER / PRIMARY CULTURE / ORGANOTYPIC CULTURE / 一酸化窒素生成酵素(N0S) / 脳領野 / 小脳 organotypic culture / 一酸化窒素生成酵素 / 小脳顆粒細胞初代培養 / Micro-Western blot / nNOS mRNA / nNOS / 一酸化窒素生成酵素同位体 / 定量的RT-PCR / 栄養因子 / 小脳顆粒細胞
Research Abstract

Nitric oxide synthase (NOS), which catalyzes the formation of the information molecule nitric oxide (NO), is widely distributed in general tissue cells and its high concentrations are found in the cerebellar granular layr. Cerebellar granule cells differentiate postnatally and the cerebellar layrs are known to develop accordingly. Thus, in order to investigate the differentiation and maturation process of granule cells in the postnatal development of rat cerebellum, NOS activities and isoforms were analyzed in the developing layrs. We developed a microassay method for NOS,using an enzymatic amplification reaction NADP cycling. NOS activities were increased in the molecular and internal granular layrs during development and the activity was detected in the external granular layrs. Our newly developed micro-immunoblot showed neuronal NOS (nNOS) and endothelial NOS (eNOS) in the molecular layr and internal granular layr, and the nNOS immunostaining intensity increased during the postnatal development, representing differentiation and maturation in both layrs. eNOS and nNOS were stained in immunoblot of the neuroblastic external granular layr. Further-more, nNOS mRNA was also detected in this germinal layr, using our newly devised micro-immunoblot. Intraperitoneal administration of N^G-nitro-L-arginine inhibited NOS activity in vivo and disturbed the layr formation in developing cerebellum ; thus NO seems to support the differentiation and maturation of granule cells. In the primary culture of granules cells, isolated from the postnatal day 9 cerebella, NOS activity and nNOS mRNA increased during maturation in vitro. The organotypic culture of the developing slices, prepared from the postnatal day 9 cerebella, showed migration and maturation of granule cells to from the cerebellar layrs, but the layr structure was irregular and the preparation of good microsamples form the slices was hardly possible.

Report

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

    (45 results)

All Other

All Publications (45 results)

  • [Publications] Wang W, Nakayama T, Inoue N, Kato T: "Quantitative analysis of nitri oxide synthase expressed in developing and diffettrentiating rat cerebellum." Developmental Brain Rsearch. in press (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato T, Momoki T, Nakayama T, Inoue N, Sato S, Sekihara H: "Expression of glucose transporter isoforms,GLUT1 and GLUT3,in developing rat cerebellum." Journal of Neurochemistry. 69. S25 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Furuya S, Nagata R, Ozaki Y, Furuya K, Nakayama T, Nagahama M: "A monocolnal antibody to astrocytes,subepithelial fobloblasts of small intestial villi and interstitial cells of the myentric plexus layer" Anatomical Embryology. 195. 113-126 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagamatsu, S., Nakamichi, Y., Inoue, N., Inoue, M., Nishino, H., Sawa, H.: "Rat 6 glioma cell growth is related to glucose transport and metabolism." Biochemical Journal. 319. 477-482 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wang, W., Inoue, N., Nakayama, T., Ishii, M, Kato, T.: "Assay method for nitric oxide synthase in crude samples determining product NADP^+." Analytical Biochemistry. 227. 274-280 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nomoto, M., Gonzalez, F.J., Mita, T., Inoue, N., Kawamura, M.: "Analysis of cis-acting regions upstream of the rat Na^+/K^+-ATPase α1 subunits gene by in vivo footprinting." Biochimica Biophysica Acta. 1264. 35-39 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato, T., Kijima.M., Ohno, S.: "Distribution of S-antigen and its behavior in ocular tissues of control and experimental autoimmune uveoretinitis rats. Retinal Degeneration and Regeneration (eds.Kato,S.,Osborne,N.N.,Tamai,M.)" Kugler Publ.Amsterdam/New York, 99-104 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 加藤 尚彦: "NOと中枢神経(NOの基礎と臨床:室田誠逸編)" メディカルレビュー社, 186-214 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Sasaki, R., Nakayama, T., Kato, T.: "Protein expression patterns in mouse oocytes and preimplantation embryos." Devel.Biol.(submitted.). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wang, W., Nakayama, T., Inoue, N., Kato, T.: "Expression of nitric oxide synthase in developing and differentiating rat cerebellum." Dev.Brain Res.(in press.). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Furuya, S., Nagata, R., Ozaki, Y., Furuya, K., Nakayama, T., Nagahama, M.: "A monoclonal antibody to astrocytes, subepithelial fibroblasts of small intestinal villi and interstitial cells of myenteric plexus." Anat.Embyol.195. 113-126 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato, T., Momoki, T., Nakayama, T., Inoue, N., Sato, S., Sekihara, H.: "Expression of glucose transporter isoforms, GLUT1 and GLUT3, in developing rat cerebellum." J.Neurochem. 69 : (Suppl.). S25 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yamaguchi, K., Nakayama, T., Fujisawa, T., Akagawa, K.: "Enhancement of neurite-sprouting by suppression of HPC-1/syntaxin 1A activity in cultured vertebrate nerve cells." Brain Res.740. 185-192 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato, T., Inoue, N., Nakayama, T.: "Expression of nitric oxide synthase (NOS) in developing and maturing rat cerebellum." Bulletin of the Japanese Society for Neurochemistry. 35. 218-219 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nagamatsu, S., Nakamichi, Y., Inoue, N., Inoue, M., Nishino, H., Sawa, H.: "Rat C6 glioma cell growth is related to glucose transport and metabolism." Biochem.J.319. 477-482 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wang, W., Inoue, N., Nakayama, T., Ishii, M., Kato, T.: "Assay method for nitric oxide synthase in crude samples determining product NADP^+." Anal.Biochem.227. 274-280 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Nomoto, M., Gonzalez, F.J., Mita, T., Inoue, N., Kawamura, M.: "Analysis of cis-acting regions upstream of the rat Na^+/K_+-ATPase alpha1 sub-units gene by in vivo footprinting." Biochim.Biophys.Acta.1264. 35-39 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wang, W., Inoue, N., Nakayama, T., Ishii, M., Kato, T.: "Nitric oxide synthase (NOS) supports development and differentiation of neurons in cerebellar layrs from rats." J.Neurochem.65. S215A (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Inoue, N., Soga, T., Nakayama, T., Kato, T.: "Expression and regulation of sodium pump isoforms in cultured cerebellar granule cells." J.Neurochem.65. S159D (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato, T., Kijima, M.: "Microanalysis of S-antigen distribution in ocular tissues and retinas from rats." Neurochem.Res.20. 331 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wang, W., Inoue, N., Nakayama, T., Ishii, M., Kato, T.: "Nitric oxide synthase (NOS) activities in developing and differentiating cerebellar layrs from rats." Neurochem.Res.20. 365 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kato, T., Kijima, M., Ohno, S.: Distribution of S-antigen and its behavior in ocular tissues of control and experimental autoimmune uveoretinitis rats. Retinal Degeneration and Regeneration. Kugler Publ.Amsterdam/New York., 99-104 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Wang W,Nakayama T,Inoue N,Ishii M,Kato T: "Quantitative analysis of nitric oxide synthase expressed in developing and diffettrentiating rat cerebellum." Developmental Brain Rsearch. (in press). (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] Furuya S,Nagata R,Ozaki Y,Furuya K,Nakayama T,Nagahama M: "A monocolnal antibody to astrocytes,subepithelial fobloblasts of small intestial villi and interstitial cells of the myentric plexus layer" Anatomical Embryology. 195. 113-126 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kato T,Momoki T,Nakayama T,Inoue N,Sato S,Sekihara H: "Expression of glucose transporter isoforms,GLUT1 and GLUT3,in developing rat cerebellum" Journal of Neurochemistry. 69. S25 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kato T,Inoue N,Nakayama T: "Expression of nitric oxide synthase in developing and differentiating rat cerebellum." Neurochemical Research. 22. 850 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Inoue N,Soga T,Nakayama T,Kato T: "Expression of nitric oxide synthase during development of culatured rat cerebellar granule cells." Neurochemical Research. 22. 890 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 井上順雄,曽我智子,原諭吉,中山孝,加藤尚彦: "遺伝子導入細胞を用いた神経細胞型Naポンプ活性調節機構の解析" 神経化学. 36. 285 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 井上 順雄: "陽イオンポンプ:脳における情報伝達;神経機能素子と素過程(蛋白質核酸酵素増刊)" 共立出版, 235-243 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kato T,Inoue N,Nakayama T: "Expression of nitric oxide synthase (NOS) in developing and maturing rat cerebellum." Bulletin of the Japanese Society for Neurochemistry. 35. 218-219 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Nagamatsu S,Nakamichi Y,Inoue N,Inoue M,Nishino H,Sawa H: "Rat C6 glioma cell growth is related to glucose trasport and metabolism." Biochem.J.319. 447-482 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Yamaguchi K,Nakayama T,Fujiwara T,and Akagawa K: "Enhancement of neurite-sprouting by suppression of HPC-1/syntaxin 1A activity in cultured vertebrate nerve cells." Brain Res.740. 185-192 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 百木忠久,中山孝,井上順雄,佐藤忍,関原久彦,加藤尚彦: "分化発達するラット小脳におけるGlucose Transporter (GLUT) 1と3の発現." 神経化学. 35. 372-373 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 井上順雄,曽我智子,中山孝,加藤尚彦: "培養小脳顆粒細胞の発達に伴う一酸化窒素合成酵素の発現" 神経化学. 35. 558-559 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 中山孝,五嶋良郎,加藤尚彦,三須良實: "ニワトリ視蓋反発因子の細胞内情報伝達機構の解析" 神経化学. 35. 594-595 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kato T,Kijima M,Ohno S.: "Distribution of S-antigen and its behavior in ocular tissues of control and EAU (Experimental Autoimune Uveoretinitis) rats,in RETINAL REGENERATION" Kuluger Publications bv,Amsterdam,the Nitherland, 99-104 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] 加藤尚彦: "NOと中枢神経(NOの基礎と臨床:室田誠逸編)" メディカルレビュー社, 186-214 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Wang W,Inoue N,Nakayama T,Ishii M, Kato T.: "Assay method for nitric oxide synthase in crude samples determining product NADP^+." Analytical Biochemistry. 227. 274-280 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wang W,Inoue N,Nakayama T,Ishii M, Kato T.: "Nitric oxide synthase (NOS)supports development and differentiation of neurons in erebellar layers from rats." Journal of Neurochemistry. 65. S215A (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Inoue N,Soga T,Nakayama T,Kato T.: "Expression and regulation of sodium pump isoforms in cultured cere-bellar granule cells." Journal of Neurochemistry. 65. S159D (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kato T,Kijima M.: "Microanalysis of S-antigen distribution in ocuar tissues and retinas from rats." Neurochemical Reseach. 20. 331 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wang W,Inoue N,Nakayama T,Ishii M, Kato T.: "Nitrix oxide synthase (NOS)activities in developing and differentiating cerebellar layers from rats." Neurochemical Reseach. 20. 365 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Inoue N,Soga T,Nakayama T,Kato T.: "Sodium pump isoforms in cultured granule cells." Neurochemical Reseach. 20. 334 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kato T, Kijima M, Ohno S.: "Disribution of S-antigen and its behavior in ocular tissues of control and EAU (Experimental Automune Uveoretinitis)rats,in RETINAL REGENERATION" Kuluger Publications bv,Amsterdam,the Nitherland(in press), (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 加藤 尚彦: "NOと中枢神経(NOの基礎と臨床:室田誠逸編)" メディカルレビュー社(in press), (1996)

    • Related Report
      1995 Annual Research Report

URL: 

Published: 1996-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi