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The study toward the development of specific agonist and antagonist for the purinergic receptors in the kidney

Research Project

Project/Area Number 08672623
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用薬理学・医療系薬学
Research InstitutionKyorin University

Principal Investigator

SEKINE Takashi  Kyorin University, School of Medicine, Research Associate, 医学部, 助手 (50255402)

Co-Investigator(Kenkyū-buntansha) HOSOYAMADA Makoto  Kyorin University, School of Medicine, Research Associate, 医学部, 助手 (00291659)
TAKEDA Michio  Kyorin University, School of Medicine, Assistant Professor, 医学部, 講師 (40255401)
ENDOU Hitoshi  Kyorin University, School of Medicine, Professor, 医学部, 教授 (20101115)
CHA Seok ho  Kyorin University, School of Medicine, Research Associate, 医学部, 助手 (50276200)
Project Period (FY) 1996 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Keywordskidney / purinoceptor / intracellular calcium / nephron segments / ATP / proximal tubules / mesangial cells / purinoreceptor / organic anion transporter / protein kinase C / phosphorylation / nephron / calcium / P_<2Y> / P_<2U> / 尿細管 / 細胞内カルシウム / desensitization
Research Abstract

During the research performed from 1996 to 1998, we obtained the following results. In order to determine the existence of the ATP receptors along nephron segment, we investigated a [CaィイD22+ィエD2]I increase induced by ATP and its analogs in isolated single nephron segment. An increase of [CaィイD22+ィエD2]I was observed in the glomerulus, proximal tubule and outer medullar collecting duct (OMCD), and the subtypes were determined to be P2Y1 of P2Y2. After being treated with ATP, a [CaィイD22+ィエD2]I increase was not observed by addition of several peptide hormones, such as AVP and ANG II, in the nephron segments. This result suggests that ATP receptor might modulate the renal effect of these hormones.
We also investigated the existence of ATP receptors in the renal cultured cells which we established from the distinct nephron segments derived from SV40 large T antigen transgenic mouse. The mRNA expression of P2X and P2Y purinoceptors were detected in the culture cells of mouse S1 (early convoluted segment of proximal tubule) and OMCD. Addition of ATP and its analogs induced the [CaィイD22+ィエD2]I increase in both of the cells.
In addition, rat cultured mesangial cells expressed the P2Y and P2X purinoceptors, and showed the [CaィイD22+ィエD2]I increase induced by ATP. Phosphorylation of myosin light chain was not detected by the stimulation of ATP receptors of mesangial cells.
Thus, this project revealed the existence of certain subtypes of purinoceptors in the kidney and renal culture cells. We also implicated several physiological roles of purinoceptors of the kidney. We hope that the findings obtained in this project will promote the understanding the physiological significance of purinoceptors, and provide the basis on the development of agonist and antagonist of the purinoceptors.

Report

(4 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • 1996 Annual Research Report
  • Research Products

    (20 results)

All Other

All Publications (20 results)

  • [Publications] Sekine,T. et. al.: "Expression cloning and characterization of a novel multispecific organic anion trasporter."J. Biol. Chem.. 272. 18526-18529 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sekine,T. et. al.: "Cloning,functional characterization and localization of a rat renal Na^+-dicarboxylatetransporter."Am. J. Physiol.. 275. F298-305 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sekine,T. et. al.: "Identification of multispecific organic anion transporter 2 expressed predominantly in the liver."FEBS letter. 429. 179-182 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Cha,SH.,Sekine,T. et. al.: "P2-purinoreceptor localization along a rat nephron and evidence suggesting the existence of subtype,P2Y1 and P2Y2."Am. J. Physiol.. 274. F1006-1014 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sekine,T. et. al.: "Molecular cloning and characterization of a high-affinity carnitine transporter from rat intestine"Biochem. Biophys. Res. Commun.. 251(2). 586-591 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kusuhara,H.,Sekine,T. et. al.: "Molecular cloning and characterization of a new multispecific organic anion transporter from rat brain."J. Biol. Chem.. 274. 13675-13680 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sekine, T. et. Al.: "Expression cloning and characterization of a novel multispecific organic anion transporter"J. Biol. Chem.. 272(30). 18526-18529 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Cha, SH., Sekine, T. and Endou, H.: "P2-purinoreceptor localization along a rat nephron and evidence suggesting the existence of subtype, P2Y1 and P2Y2"Am. J. Physiol.. 274. F1006-1014 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takeda, M., Kobayashi, M. and Endou, H.: "Establishment of mouse clonal early proximal tubule cell line and outer medullary collecting duct cells expressing P2 purinoceptors"Biochem. Mol. Biol. Int.. 44(4). 657-664 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sekine, T., et. Al.: "Cloning, functional characterization and localization of a rat renal Na+-dicarboxylate transporter"Am. J. Physiol.. 275. F298-305 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sekine, T., et al.: "Identification of multispecific organic anion transporter 2 expressed predominantly in the liver"FEBS letter. 429. 179-182 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sekine, T., et al.: "Molecular cloning and characterization of a high-affinity carnitine transporter from rat intestine"Biochem. Biophys. Res. Commun.. 251(2). 586-591 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kusuhara, H., Sekine, T., et al.: "Molecular cloning and characterization of a new multispecific organic anion transporter from rat brain"J. Biol. Chem.. 274. 13675-13680 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sekine,T.et al.: "Cloning,functional characterization and localization of a rat renal Na^+-dicarboxylatetransporter." Am.J.Physiol.275. F298-305

    • Related Report
      1998 Annual Research Report
  • [Publications] Sekine,T.et al.: "Identification of multispecific organic anion transporter 2 expressed predominantly in the liver." FEBS letter. 429. 179-182

    • Related Report
      1998 Annual Research Report
  • [Publications] Cha,SH.,Sekine,T.et al.: "P2-purinoreceptor localization along a rat nephron and evidence suggesting the existence of subtype,P2Y1 and P2Y2." Am.J.Physiol.274. F1006-1014

    • Related Report
      1998 Annual Research Report
  • [Publications] Sekine,T.,et al.: "Molecular cloning and characterization of a high-affinity carnitine transporter from rat intestine." Biochem.Biophys.Res.Commun.251(2). 586-591

    • Related Report
      1998 Annual Research Report
  • [Publications] Tojo,A.,Sekine,T.,et al.: "Immunohistochemical localization of multispceicif renal organic anion transporter(OAT1)in rat kidney." J.Am.Soc.Nephrol.(in press).

    • Related Report
      1998 Annual Research Report
  • [Publications] Kusuhara,H.,Sekine,T.,et al.: "Molecular cloning and characterization of a new multispecific organic anion tranporter from rat brain." J.Biol.Chem.(in press).

    • Related Report
      1998 Annual Research Report
  • [Publications] Cha S.H., Sekine T.,et al.: "P2-purinoreceptor localization along a rat nephron and evidence........" American Journal Physiology. (in press).

    • Related Report
      1997 Annual Research Report

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

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