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Analysis of mechanisms regulation ion transport activities of Na pump isoforms in neurons.

Research Project

Project/Area Number 13680847
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Neurochemistry/Neuropharmacology
Research InstitutionYokohama City University

Principal Investigator

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

Co-Investigator(Kenkyū-buntansha) NAKAYAMA Takashi  Yokohama City University School of Medicine, Biochemistry, Lecturer, 医学部, 講師 (90150060)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2001: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsNeuron / Na pump / Na, K-ATPase / Isoform / Ion transport / ES cell / Differentiation / Na, K-ATPase / 細胞外カリウム / リン酸化
Research Abstract

Three Na pump isoforms α1, α2 and α3 isoforms, are present in cultured neurons and ion transport activities of the isoforms are differentially regulated. Here we show that the inhibition of α2/α3, a neuron type Na pump, isoform by extracellular K^+ is a key mechanism of the differential regulation. Only the α2/α3 isoform is inhibited by physiological concentrations of K^+ under basal conditions in mature neurons. The inhibition was vanished after glutamate excitation of the neurons and by incubation with a calmodulin antagonist or an inhibitor of CaM kinase II. In contrast to mature neurons the inhibition is small in immature neurons, in which the isoform activities are not regulated differentially. Additionally, when ATP concentration in the neurons becomes low, the inhibition diminishes and the differential regulation becomes increasingly prominent. We also show that astrocyte-derived factors instruct mouse ES cells to differentiate into neurons. Cultured in astrocyte-conditioned med … More ium (ACM) under free-floating conditions, within 4 days, colonies of undifferentiated ES cells give rise to floating spheres with a concentric stratiform structure. Specifically, the spheres have a periphery of neural stem cells, a core of dividing ES cells and an intermediate layer. Culturing the spheres on an adhesive substrate in ACM promotes neurogenesis. The attached spheres give rise to clusters of cells containing neurons at the periphery, and many neurons with neurites migrate from the clusters within 5 days. Gene expression analyses and electrophysiology confirm the neuronal properties of the cells. The neurons express a neuron type Na pump, the α2 and α3 isoforms. Immunofluorescence analyses show that many NF^+ neurons and neurites are also positive for tyrosine hydroxylase. In contrast, neither astrocytes nor oligodendrocytes are formed. This pathway provides a new insight into mechanisms of neurogenesis and also provides a simple procedure for generation of neural stem cells and neurons. Less

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] Soga, M.T.: "Expression and regulation of Na pump isoforms in cultured cerebellar granule cells"Neuroreport. 12. 829-832 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inoue, N.: "Participation of extracellular potassium and intracellular sodium in regulation of sodium pump isoform activities in neurons"Neurochem. Res.. 26. 299-300 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kato, T.: "Quantitative analysis of nitric oxide synthase (NOS) mRNA and protein in the amygdala and hippocampus of fear-conditioned rat"Neurochem. Res.. 26. 320 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Cheng, C.: "Cdk/p35-dependent and independent pathways through which eprin-A5 induces morphological changes of neuronal and non-neuronal cell"Neurosci. Res.. Suppl.25. S51 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sasaki, Y.: "Semaphorin-3A signaling is mediated with Fyn-Cdk5 phosphorylation cascade"Neurosci. Res.. Suppl.25. S51 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inoue, N.: "Mechanism of regulation of ion transport activity of a neuron-type Na pump"Neurochem. Res.. 26. 1355 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sasaki, Y.: "Fyn and cdk5 mediate Semaphorin-3A signaling, which is involved in regulation of dendrite orientation in cerebral cortex"Neuron. 35. 907-920 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakayama, T.: "A direct differentiation pathway from murine embryonic stem cells to neurons"Abstract for Keystone Symposia on From Stem Cells to Therapy. 71 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakayama, T.: "Astrocyte-derived factors instruct differentiation of embryonic stem cells into neurons"Neurosci. Res.. 41. 241-249 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakayama, T.: "Embryonic stem cells directly differentiate into neurons"Neurosci. Res.. Suppl.27(印刷中). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Soga-Momoki T, Nakayama T and Inoue N: "Expression and regulation of Na pump isoforms in cultured cerebellar granule cells."NeuroReport. 12. 829-832 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inoue N, Soga T and Kato T: "Participation of extracellular potassium and intracellular sodium in regulation of sodium pump isoform activities in neurons."Neurochem Res. 26. 299-300 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Kato T, Osakada K, Sato K, Abe K, Inoue N and Nakayama T: "Quantitative analysis of nitric oxide synthase (NOS) mRNA and protein in the amygdala and hippocampus of fear-conditioned rat."Neurochem Res. 26. 320 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Cheng C, Sasaki Y, Nakayama T, Tanaka H, Ohno S and Goshima Y: "Cdk/p35-dependent and independent pathways through which eprin-A5 induces morphological changes of neuronal and non-neuronal cells."Neurosci Res. Suppl. 25. S51 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sasaki Y, Cheng C, Uchida Y, Ohshima T, Yagi T, Taniguchi M, Nakayama T, Kudo Y, Ohno S, Nakamura F and Goshima Y: "Semaphorin-3A signaling is mediated with Fyn-Cdk5 phosphorylation cascade."Neurosci Res. Suppl. 25. S51 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inoue N, Soga M T & Nakayama T: "Mechanism of regulation of ion transport activity of a neurontype Na pump."Neurochem Res. 26. 1355 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sasaki Y, Cheng C, Uchida Y, Nakajima O, Ohshima T, Yagi T, Taniguchi M, Nakayama T, Kishida R, Kudo Y, Ohno S, Nakamura F and Goshima Y: "Fyn and cdk5 mediate Semaphorin-3A signaling, which is involved in regulation of dendrite orientation in cerebral cortex."Neuron. 35. 907-920 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakayama T, Momoki-Soga T, Yamaguchi K, Suzuki Y, Kondo Y and Inoue N: "A direct differentiation pathway from murine embryonic stem cells to neurons."Abstract for Keystone Symposia on From Stem Cell to Therapy. 71. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakayama T, Momoki-Soga T & Inoue N: "Astrocyte-derived factors instruct differentiation of embryonic stem cells into neurons."Neurosci Res(in press). 41(2). 241-249 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Nakayama T, Momoki-Soga T, Yamaguchi K, Suzuki Y, Kondo Y, Inoue N: "Embryonic stem cells directly differentiate into neurons."Neurosci Res(in press). Suppl. 27. (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Sasaki, Y.: "Fyn and cdk5 mediate Semaphorin 3A signaling, which is involved in regulation of dendrite orientation in cerebral cortex"Neuron. 35. 907-920 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nakayama, T.: "Astrocyte-derived factors instruct differentiation of embryonic stem cells into neurons"Neurosci Res. 41(印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nakayama, T.: "A direct differentiation pathway from murine embryonic stem cells to neurons"Abstract of Keystone Symposia on From Stem Cells to Therapy. (印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Nakayama, T.: "Embryonic stem cells directly differentiate into neurons"Neurosci Res. Suppl.26(印刷中). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Inoue, N.: "Regulation of sodium pump isoforms after glutamate excitation of cultured cerebral neurons"Neurosci.Res.. 24. S61 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kato, T.: "Expression of nitric oxide synthase (NOS) isoforms in the amygdala and hippocampus of fear-conditioned rat"Neurosci.Res.. 24. S115 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Soga, M.T.: "Expression and regulation of Na pump isoforms in cultured cerebellar granule cells"Neuroreport. 12. 829-832 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Inoue, N.: "Participation of extracellular potassium and intracellular sodium in regulation of sodium pump isoform activities in neurons"Neurochem.Res.. 26. 299-300 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Kato, T.: "Quantitative analysis of nitric oxide synthase (NOS) mRNA and protein in the amygdala and hippocampus of fear-conditioned rat"Neurochem.Res.. 26. 320 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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