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Roles of cloned chloride channels in kidney chloride transport

Research Project

Project/Area Number 07671241
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Kidney internal medicine
Research InstitutionTokyo Medical and Dental University

Principal Investigator

UCHIDA Shinichi  Tokyo Medical and Dental University, School of Medicine, Instructor, 医学部, 助手 (50262184)

Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordschloride channel / immunohistochemistry / human gene / FISH method / イオントランスポート / 尿濃縮 / 腎臓
Research Abstract

To determine the intrarenal localization of three chloride channels isolated by us, we prepared antisera for rat ClC-K1, -K2, -3 using synthetic peptide as antigens. We obtained specific antisera to ClC-K1 and ClC-K2. Using these antisera, we performed immunohistochemical study to localize the intrarenal localizaton of ClC-K1 and ClC-K2. ClC-K1 is present in the both apical and basolateral plasma membranes of the thin ascending limb of Henle's loop, and ClC-K2 is present in the apical plasma membrane of the connecting tubules. These findings will shed light on the physiological roles if ClC-K1 and -K2 in the kidney.
Next, we studied structure- function relationship of ClC-K1 and ClC-3. The in vitro mutations of positively charged amino acids in the first extracellular loop of ClC-K1 into negatively charged amino acids greatly enhanced the chloride current expressed in Xenopus oocytes, suggesting the importance of this region. Further studies using chimera chloride channels of ClC-K1 and -K2 are in progress. As for ClC-3, a stably expressing cell line was established in CHO cells, and patch clamp analysis revealed the Ca-regulated nature of ClC-3. Finally, we isolated human ClC-K1 and -K2 cDNAs to study the involvements of these genes in certain human kidney diseases. Using these cDNA,we further isolated human ClC-K1 and -K2 genes and characterized the gene structure including exon-intron structures. We prepared PCR primer sets to cover all exons and exon-intron boundaries and could established the system to determine the sequences of ClC-K2 gene from patients by direct sequencing. Also, we determined the chromosomal localization of ClC-K1, -K2 and -3 by FISH method.

Report

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

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Yasuo Takeuchi,Shinichi Uchida,Fumiaki Marumo,Sei Sasaki: "Cloning, tissue distribution, and intrarenal localization of CIC chloride channels in humon kidney" Kidney International. 48. 1497-1503 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Masanobu Kawasaki,M.Suzuki,Shinichi Uchida,et al: "Stable and functional expression of CIC-3 chloride channel in somatic cell lines" Neuron. 14. 1285-1291 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shinichi Uchida,Sei Sasaki, et al: "Locolfzation and functional characterization of rat kidney-specific chloride channel, CIC-K1" Journal of Cln'cal Investigation. 95. 104-113 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shinichi Uchida, Sei Sasaki,Fumiaki Marumo: "Chloride transport across kidney epithelia throogh CIC chloride channel" Japanese Journal of Nephlorogy. 38. 285-289 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Shinichi Uchida,Fumiaki Marumo: "Molecular characterization of chloride channels in the kidney" Experimental Neghrogy. 4. 135-138 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Fumiko Saito-Ohara,Shinichi Uchida et al: "Assignment of the genes ancoding the human chloride channels, CLCKA and CLCKB, to 2p36 and of cLCN3 to 4q32-33 by Insitu hybridization" Genomics. 36. 372-374 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Saito-Ohara F,Uchida S,TakeuchiY,Sasaki S,Hayashi A,Marumo F,Ikeuchi T: "Assignment of the genes encoding the human chloride channels, CLCNKA and CLCNKB,to 1p36 and of CLCN3 to 4q32-q33 by in situ hybridization." Genomics. 36. 372-374 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Furuno M,Uchida S,Marumo F,Sasaki, S.: "Repressive regulation of the aquaporine-2 gene." Am J Physiol Renal, Fluid Electrolyte Physiol. 271. F854-F860 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Rai T,Uchida S,Sasaki S,Marumo F: "Footprint analysis of rat aquaporin-2 (AQP-2) gene : implication of a novel transcription factor binding site" J Am Soc Nephrol. 6. 328 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uchida S,Marumo F: "Molecular characterization of chloride channel in the kidney." Exp Nephrol. 4. 135-138 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uchida S,Sasaki S,Marumo F: "Chloride transport across kidney epithelia through ClC chloride channels." Jpn J Nephrol. 38. 285-289 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uchida S,Sasaki S,Nitta K,Uchida K,Horita S,Nihei H,Marumo F: "Localization and functional characterization of rat kidney-specific chloride channel, ClC-K1." J Clin Invest. 95. 104-113 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takenaka M,Bagnasco SM,Preston AS,Uchida S,Yamauchi A,Kwon HM Handler JS: "The canine betaine gamma-amino-n-butyric acid transporter gene : Diverse mRNA isoforms are regulated by hypertonicity and are expressed in a tissue-specific manner" Proc Natl Acad Sci USA. 92. 1072-1076 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kawasaki M,Suzuki M,Uchida S,Sasaki S,Marumo F: "Stable and functional expression of the ClC-3 chloride channel in somatic cell lines" Neuron. 14. 1285-1291 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Imase N,Fushimi K,Ishibashi K,Uchida S,Ichioka M,Sasaki S,Marumo F: "Isolation of human aquaporin 3 gene" J Biol Chem. 270. 17913-17916 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Takeuchi Y,Uchida S,Marumo F,Sasaki S: "Cloning, tissue distribution, and intarenal localization of ClC chloride channels in human kidney." Kidney Int. 48. 1497-1503 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Sakamoto H,Kawasaki M,Uchida S,Sasaki S,Marumo F: "Identification of a new outwardly rectifying Cl-channel that belongs to a subfamily of the ClC Cl-channels." J Biol Chem. 271. 10210-10216 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Matsumura Y,Uchida S,Sasaki S,Marumo F: "Identification of the hypertonicity-responsive element in human Aquaporin-2 promoter." J Am Soc Nephrol. 6. 326 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Uchida S,Sasaki S,Marumo F: Functional regulation of CIC-K1 chloride channel by protein kinase A ; in Saito T,Kurokawa K,Yoshida S (eds) : Neurohypophysis : recent progress of vasopressin and oxytocin research.Amsterdam, Elsevier, 545-550 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Yasuo Takeuchi,Shinichi Uchida,Fumiaki Marumo,Sei Sasaki: "Cloning,tissue distribution,and intrarenol localization of CIC chloride channels in human kidney" Kidney International. 48. 1497-1503 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Masanobu Kawasaki,Makoto Suzuki Shinichi Uchida,Sei Sasaki,Fumiaki Marumo: "Stable and functronal expression of CIC-3 chloride channel in somatic cell ilnes" Neuron. 14. 1285-1291 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Shinichi Uchida,Sei Sasaki Fumiaki Marumo et.al.: "Localization and functional characterization of rat kidney-specific chloride channel,CIC-K1" Journal of Clinical Investigation. 95. 104-113 (1995)

    • Related Report
      1996 Annual Research Report
  • [Publications] Shinichi Uchida,Sei Sasaki Fumiaki Marumo: "Chloride transport across kidney epithelia through CIC chloride channel" Japanese Journal of Nephrology. 38. 285-289 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Shinichi Uchida Fumiaki Marumo: "Molecular characterization of Chloride Channels in the kidney" Exp.Nephrology. 4. 135-138 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Fumiko Saito-Ohara,Shinichi Uchida et.al: "Assignment of the genes encoding the human chloride channels,CLCKA and CLCKB,to Ip36 and of CLCN3 to 4q32-q33 by in situ hybridization" Genomics. 36. 372-374 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Uchidas.,et al: "Locolization and functional characterization of rat kidney-specific chloride channel,CIC-K1" J.Clin.Invest.95. 104-113 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Takeuchi Y.,Uchida S.,et.al: "Cloning,tissue distribation,andintrd renal locolization of CIC-chloride channel in human kidney" Kidney International. 48. 1497-1503 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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