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Analysis of regulational mechanism of Na pump isoform activities using cells transfected neuron-type isoform.

Research Project

Project/Area Number 09680768
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, Department of Biochemistry, Associate professor, 医学部, 助教授 (50159985)

Co-Investigator(Kenkyū-buntansha) HARA Yukichi  Tokyo Medical and Dental University Medical Research Division, Department of Ret, 大学院医学研究科, 助教授 (00092429)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1998: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1997: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsNa, K-pump / Na, K-ATPase / isoform / neuron / regulation / gene transfection
Research Abstract

Na pump isoforms, "common-type (alpha)" and "neuron-type (alpha2 and alpha3)" isoforms, were expressed in cultured rat cercbral neurons. When the cells were excited with glutamate or maintained in energy-deficient states, the K^+ uptake activities of Na pump isoforms were regulated differently : that of neuron-type isoform increased but that of alpha isoform decreased. To analyze mechanism(s) of the differential regulation of K^+ uptake activities of the isoforms in the neurons, we used T3-3-3 cells which were established by transfecing alpha3 subunit cDNA into BALB/c 3T3 cells and expressed both preexisting alpha1 isoform and transfected alpha3 isoform, In the IC uptake activity of the cells, affinity of a3 isoform for intracellular Na^+ was lower than that of alpha1 isoform. Patch-clamp technique demonstrated that pump current of alpha3 isoform was activated by membrane depolarization but that of alpha1 isoform was not. These findings suggest that the differential regulation of K^+ uptake activities of the isoforms after excitation with glutamate results from the difference in pump kinetics between the isoforms. However, the inhibition of the K^+ uptake activity of alpha1 isoform after excitation occurred in mature neurons, in contrast activation of the activity occurred in immature neurons. Therefore, some mechanism(s) in intracellular signal transduction may also be involved in the differential regulation and developed during maturation in culture. To analyze the mechanism, the T3-3-3 cells were treated with dibutyryl cAMP or forskolin or PMA, however, no or less effect on the isoform activities was detected. Affinity of alpha3 isoform for ATP was smaller than that of alpha1 isoform in the ATPase activity of the T3-3-3 cells. Therefore, it is speculated that a part of the regulation of the Na pump isoform activities in energy-deficient states of the neurons is due to differences in affinities for ATP between the isoforms.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (37 results)

All Other

All Publications (37 results)

  • [Publications] 井上順雄: "陽イオンポンプ" 蛋白質核酸酵素. 42(3). 39-47 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kato, T.: "Expression of glucose transporter isoforms, GLUT1 and GLUT3, in developing rat cerebellum." J Neurochem. 69. S25 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kato, T.: "Expression of nitric oxide synthase in developing and differentiating rat cerebellum." Neurochem Res. 22(7). 850 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Momoki, T.: "Expression of glucose transporter isoforms, GLUT1 and GLUT3, in developing rat cerebellum." Neurochem Res. 22(7). 868 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Inoue, N.: "Expression of nitric oxide synthase during development of cultured rat cerebellar granule cells." Neurochem Res. 22(7). 890 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Urayama, O.: "Coincident induction of Krev-1/rap1A, rap1B and H-ras mRNAs in the rat spinal cord by noxious stimulation." Mol Brain Res. 45. 331-334 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Wang, W.: "Quantitative analysis of nitric oxide synthase expressed in developing and differentiating rat cerebellum." Dev Brain Res. 111(1). 65-75 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shima, Y.: "A Missense Mutation of the gene for Na^+, K^+-ATPase α-subunit causes abnormal feeding behavior in Canorhadbitis elegans." Biochem Biophys Res Commun. 248. 778-782 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawai, M.: "A family with high serum leucine aminopeptidase (EC 3.4.11.2) activity derived from a novel variant CD13." Clin Chem. 44(2). 215-220 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Hara, Y.: "Different pump kinetics between α1 and α3 isoform of NaK-ATPase." Neurochem Res. 24(1). 179 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Inoue, N.: "Regulation of Na pump isoform activities in transfected cells expressing neuron-type α3 isoform." Neurochem Res. 24(1). 179-180 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Aisaki, K.: "Apoptotic changes precede mitochondrial dysfunction in red cell-type pyruvate kinase mutant mouse erythroleukemia cell lines." Jpn J Cancer Res. 90. 171-179 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Abe, K.: "Messenger RNA of neuronal nitric oxide synthase is expressed and possibly functions in mouse oocytes and embryo during preimplantation development." Biomed Res. 20(1) in press. (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 井上順雄: 印刷中, (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Inoue N.: "Reguration of cation pump activity in nervous system" Protein Nucleic Acid Enzyme. 42. 39-47 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kato T., Inoue N.and Nakayama T.: "Expression of nitric oxide synthase in developing and differentiating rat cerebellum" Neurochem.Res.22. 850 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Momoki T., Nakayama T., Inoue N., Sato S., Sekihara H.and Kato T.: "Expression of glucose transporter isoforms, GLUT1 and GLUT3, in developing rat cerebellum" Neurochem.Res.22. 868 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Inoue N., Soga T., Nakayama T.and Kato T.: "Expression of nitric oxide synthase during development of cultured rat cerebellar granule cells." Neurochem.Res.22. 890 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kato T., Momoki T., Nakayama T., Inoue N., Sato S.and Sekihara H.: "Expression of glucose transporter isoforms, GLUT1 and GLUT3, in developing rat cerebellum" J.Neurochem.69. S25 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Urayama O., Murakoshi T., Kubota M., Hara Y., Chihara J.And Ikawa Y.: "Coincident induction of Krev-1/rap1A, rap1B and H-ras mRNAs in the rat spinal cord by noxious stimulation" Mol.Brain Res.45. 331-334 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Wang W., Nakayama T., Inoue N.and Kato T.: "Quantitative analysis of nitric oxide synthase expressed in developing and differentiating rat cerebellum" Develop.Brain Res.111. 65-75 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shima Y., Tada Y., Furuki M., Hara Y.and Ohta H.: "A Missense Mutation of the gene for Na+, K+-ATPase alpha-subunit causes abnormal feeding behavior in Canorhadbitis elegans." Biochem.Biophys.Res.Commun.248. 778-782 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kawai M., Hara Y., Kubota T., Shiba K.and Hosaki S.: "A family with high serum leucine aminopeptidase (EC3.4.11.2) activity derived from a novel variant CD13." Clin.Chem.44. 215-220 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Hara Y.and Furukawa T.: "Different pump kinetics between alpha1 and alpha3 isoform of Na, K-ATPase." Neurochem.Res.24. 179 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Inoue N., Soga T., Hara Y., Nakayama T.and Kato T.: "Regulation of Na pump isoform activities in transfected cells expressing neuron-type alpha3 isoform" Neurochem. Res.24. 179-180 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Aisaki K., Kanno H., Oyaizu N., Hara Y., Miwa S.and Ikawa Y.: "Apoptotic changes precede mitochondrial dysfunction in red cell-type pyruvate kinase mutant mouse erythroleukemia cell lines." Jpn.J.Cancer Res.90. 171-179 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Abe K., Matsuoka K., Inoue N., Tada M.And Kato T.: "Messenger RNA of neuronal nitric oxide synthase is expressed and possibly functions in mouse oocytes and embryo during preimplantation development." Biomed.Res.20(in press). (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Wang,W.: "Quantitative analysis of nitric oxide synthase expressed in developing and differentiating rat cerebellum" Dev Brain Res. 111(1). 65-75 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shima,Y.: "A Missense Mutation of the gene for Na^+,K^+-ATPase α-subunit causes abnormal feeding behavior in Canorhadbitis elegans" Biochem Biophys Res Commun. 248. 778-782 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Inoue,N.: "Regulation of Na pump isoform activities in transfected cells expressing neuron-type α3 isoform" Neurochem Res. 24(1). 179-180 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Aisaki,K.: "Apoptotic changes precede mitochondrial dysfunction in red cell-type pyruvate kinase mutant mouse erythroleukemia cell lines" Jpn J Cancer Res. 90. 171-179 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hara,Y.: "Different pump kinetics betweenα1 and α3 isoform of NaK-ATPase" Neurochem Res. 24(1). 179 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] 井上順雄: "陽イオンポンプ" 蛋白質核酸酵素. 42(3). 39-47 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Inoue,N.: "Expression of nitric oxide synthase during development of cultured rat cerebellae granule cells" Neurochem Res. 22(7). 890 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kato,T.: "Expression of glucose transporter isoforms,GLUT1 and GLUT3,in developing rat cerebellum" J Neurochem. 69. S25 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Urayama,O.: "Coincident induction of Krev-1/rap1A,rap1B and H-ras mRNAS in the rat spinal cord by noxious stimulation" Mol Brain Res. 45. 331-334 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kawai,M.: "A family with high serum leucine aminopeptidase(EC3.4.11.2)activity derived from a novel variant CD13" Clin Chem. 44(2). 215-220 (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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