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Analysis of localization and regulation of Na, K-pump isoforms in neurons.

Research Project

Project/Area Number 06680763
Research Category

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

Allocation TypeSingle-year Grants
Research Field Neurochemistry/Neuropharmacology
Research InstitutionYokohama City University

Principal Investigator

INOUE Nobuo  Yokohama City University School of Medicine, Department of Biochemistry, Associate Professor, 医学部, 助教授 (50159985)

Co-Investigator(Kenkyū-buntansha) NAKAYAMA Takashi  Yokohama City University, School of Medicine, Department of Biochemistry, Lectur, 医学部, 講師 (90150060)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,300,000 (Direct Cost: ¥1,300,000)
KeywordsNa, K-pump / Na, K-ATPase / isoform / neuron / localization / regulation
Research Abstract

Three molecular forms of alpha (alpha1, alpha2, and alpha3) subunits of the Na, K-pump isoforms were identified in cultured granule cells, prepared from 7-day old rat cerebella, by Western blot and histochemical analysis. The subunits were shown to localize on surface of soma and neurites of the cells with ankyrin and spectrin by using confocal laser microscopy. The amounts of these subunits were shown to increase during in vitro differentiation by culture. Na, K-ATPase activities of the Na pump isoforms increased correspondingly during culture, and total activity went up to 333 (]SY.+1.[) 32 (nmol/min・mg of protein) after 13 days. The activity was 4.8-fold, higher than basal level. The brain type (alpha2 and alpha3) isoform activity increased 8.9-fold, and the common type (alpha1) only 3.4-fold. By stimulating the differentiated cells with glutamate, the K+ uptake activities of Na pump isoforms were regulated differently and total activity increased about 20%. The brain type isoform activity increased moro than fourfold by the stimulation, but the common type decreased 23%. Therefore, the percentage of the brain type isoform activity increased from 12% to 43% by the stimulation. The regulation of Na, K-Pump activities by glutamate were abolished by calmodulin antagonist W-7 and did not occur in immature neurons. Number of Na, K-pump on surface of the cells estimated by <@D13@>D1H-ouabain binding study did not change by glutamate stimulation. These results suggest that calmodulin may function in the mechanism to regulate the Na, K-pump isoform activities and that the brain type Na, K-pump isofoum in differentiated neuron is important functionally to restore the ionic gradient after excitation.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (40 results)

All Other

All Publications (40 results)

  • [Publications] Inoue,N.: "Effect of extracellular potassium concentration on sodium pump isoform activities in cultured cerebral neurons." Neurochem. Res.19. 889 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang, W.: "Microassay method for nitric oxide synthase(NOS) using an enzymatic amplification reaction NADP cycling." Neurochem. Res.19. 876 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Nagamatsu,S.: "Developmental expression of GLUT3 glucose transporter in the rat brain." FEBS Lett.346. 161-164 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Nagamatsu,S.: "Gene expression of GLUT3 glucose transporter regulated by glucose in vivo in mouse brain and in vitro in neuronal cell cultures from rat cmbryos." Biochem. J.300. 125-131 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang, W.: "An assay method for nitric oxide synthase in crude samples by determining product NADP^+" Anal. Biochem.227. 274-280 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Nomoto, M.: "Analysis of cis-acting regions upstream of the rat Na^+/K^+-ATPase α1 subunit gene by in vivo footprinting." Biochim. Biophys. Acta. 1264. 35-39 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Inoue, N.: "Sodium pump isoforms in cultured cerebellar granule cells." Neurochem. Res.20. 354 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang, W.: "Nitric oxide synthase(NOS) activities in developing and differentiating cerebellar layers from rats." Neurochem. Res.20. 365 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Inoue, N.: "Expression and regulation of sodium pump isoforms in cultured cerebellar granule cells." J. Neurochem.65. S159 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang, w.: "Nitric oxide synthase (NOS)supports development and differentiation of neurons in cerebellar layers from rats." J. Neurochem.65. S215 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 五嶋良郎: "反発因子の細胞内情報伝達" 細胞工学. 15. 181-189 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Matsui, H.: "Increase in Na pump activity of brain-type isoforms via increased turnover rate after glutamate excitation of cerebral neurons." The Sodium Pump.(Bamberg, E. & Shoner, W. eds); Steinkopff, Darmstadt, Springer, New York, (710-713)906 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] 井上順雄: "Na,K-ATPase." 生物薬科学実験講座第13巻薬物と受容体(長尾編);広川書店(印刷中), (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Nagamatsu S., Sawa H., Inoue N., Nakamichi Y., Takeshima H.and Hoshino T.18GB01 : Gene expression of GLUT3 glucose transporter regulated by glucose in vivo in mouse brain and in vitro in neuronal cell cultures from rat embryo.: Biochem.J.300. 125-131 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Nagamatsu S., Sawa H., Nakamichi Y., Katahira H.and Inoue N.: "Developmental expression of GLUT3 glucose transporter in the rat brain." FEBS Lett.346. 161-164 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang W., Inoue N., Nakayama T., Kato T.and Ishii M.: "Microassay method for nitric oxide synthase (NOS) using an enzymatic amplification reaction NADP cycling." Neurochem.Res.19. 876 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Inoue N., Matsui H.and Kato T.: "Effect of extracellular potassium concentration on sodium pump isoform activities in cultured cerebral neurons." Neurochem.Res.19. 889 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Furuya S., Nagata R and Nakayama T.: "A cytoplasmic common antigen expression in astrocytes, subepitherial fibroblasts and intestitial cells of rat small intestine." Acta Anat.Nippon. 69. 469 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Matsui H., Homareda H., Hayashi Y.and Inoue N.: "Increase in Na pump activity of brain-type isoforms via increased turnover rate after glutamate excitation of cerebral neurons." The Sodium Pump.(Bamberg, E.& Shoner, W.eds) Steinkopff, Darmstadt, Springer, New York. 710-713 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang, W., Inoue N., Nakayama T., Ishii M.and Kato T.: "An assay method for nitric oxide synthase in crude samples by determining product NADP+." Anal.Biochem.227,2. 74-280 (1995)

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

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Inoue N., Soga T., Nakayama T., and Kato T.: "Sodium pump isoforms in cultured cerebellar granule cells." Neurochem.Res.20. 354 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang W., Inoue N., Nakayama T., Kato T.and Ishii M.: "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
      1995 Final Research Report Summary
  • [Publications] Inoue N., Soga T., Nakayama T.and Kato T.: "Expression and regulation of sodium pump isoforms in cultured cerebellar granule cells." J.Neurochem.65. S159 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang W., Inoue N., Nakayama T., Ishii M.and Kato T.: "Nitric oxide synthase (NOS) supports development and differentiation of neurons in cerebellar bayrs from rats." J.Neurochem.65. S215 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Wang,W.: "Microassay method for nitric oxide synthase(NOS)using an enzymatic amplification reaction NADP cycling." Neurochem.Res.19. 876- (1994)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wang,W.: "An assay method for nitric oxide synthase in crude samples by determining product NADP^+" Anal.Biochem.227. 274-280 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Nomoto,M.: "Analysis of cis-acting regions upstream of the rat Na^+/K+-ATPase α1 subunit gene by in vivo footprinting." Biochim.Biophys.Acta. 1264. 35-39 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Inoue,N.: "Sodium pump isoforms in cultured cerebellar granule cells." Neurochem.Res.20. 354 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wang,W.: "Nitric oxide synthase(NOS)supports development and defferentiation of neurons in erebellar from rats." Neurochem.Res.20. 365- (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Inoue,N.: "Expression and regulation of sodium pump isoforms in culturedcere-bellar granule cells." J.Neurochem.65. S159 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wang,W.: "Nitric oxide synthase(NOS)supports development and differentiation of neurons in cerebellarlayers from rats." J.Neurochem.65. S215 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 五嶋良郎: "反発因子の細胞内情報伝達." 細胞工学. 15. 181-189 (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] 井上順雄: "Na,K-ATPase." 生物薬科学実験講座 第13巻 薬物と受容体(長尾編);広川書店(印刷中), (1996)

    • Related Report
      1995 Annual Research Report
  • [Publications] Wang,W.: "An assay method for nitric oxide synthase in crude samples by detemining product NADP^+" Anal.Biochem.(in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Inoue,N.: "Effect of extracellular potassium concentration on sodium pump isoform activities in cultured cerebral neurons." Neurochem.Res.19. 889- (994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Nagamatsu,S.: "Developmental expression of GLUT3 glucose transporter in the rat brain." FEBS Lett.346. 161-164 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Nagamatsu,S.: "Gene expression of GLUT3 glucose transporter regulated by glucose in vivo in mouse brain and in vitro in neuronal cell clutures from rat embryos." Biochem.J.300. 125-131 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 井上順雄: "Na,K-ATPase." 生物薬科学実験講座 第13巻 薬物と受容体(長尾編);広川書店(印刷中), (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Matsui,H.: "Increase in Na pump activity of brain-type isoforms via increased turnover rate after glutamate excitaiton of cerebral neurons." The Sodium Pump.(Bamberg,E.& Shoner,W.eds);Steinkopff,Darmstadt,Springer,New York, 906.(710-713) (1994)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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