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Gene targeting approach to study functions on respiratory and circulatory regulation of novel H^+ Channel

Research Project

Project/Area Number 11680779
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Neuroscience in general
Research InstitutionGunma University

Principal Investigator

SHIMOKAWA Noriaki  Gunma Univ School of Med, Assistant Prof, 医学部, 講師 (90235680)

Project Period (FY) 1999 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsDifferential display / Gene targeting / Ventral medullary surface / H^+-sensitivity / Hypercapnia / Rhombex-29 / 40 / MafG / CO_2 / H^+sensitivity / hypercapnia / Transmembrane Protein / H^+ sensitivity / Maf protein / proteolipid protein / Mafg / Naclear Transcription Factor / Ventral Medullary Surface / Differential Display / H^+ーSensing
Research Abstract

1. Characterization and gene targeting study of ASPT
[1] ASPT comprises 751 amino acids containing 12 membrane-spanning helices
[2] ASPT has H^+/Sugar transport motif, Proline-rich region, leucine zipper motif.
[3] ASPT immunoreactive neurons were found in the VMS and the number of cells was increased by hypercapnic stimulation.
[4] We got clones containing exons of ASPT gene from genomic library of ES cells.
[5] We constructed a vector for targeting of ASPT gene.
[6] We obtained ES cells that generated homologous recombination.
2. Detection of novel genes involved in H^+-sensitivity in the VMS
The ventral medullary surface (VMS) in known as the site of the central chemosensitive neurons. These neurons sense excess CO_2/H^+ dissolved in the cerebrospinal fluid that superfuses the VMS and induce hyperventilation. We hypothesized that genes specific for hyperventilation are expressed much more highly in VMS neurons than in extra-VMS neurons in other parts of the central nervous system (CNS). Applying the differential display technique to the brain of adult rats, we differentiated the mRNAs of the VMS neurons from those of cerebral cortex neurons.
[1] We cloned a novel four-transmembrane protein, Rhombex-29. Structural analysis and the phylogenic tree showed that rat Rhombex-29 is homologous to the major CNS myelin protein PLP/DM20-M6 family (REFERENCES 1).
[2] We cloned cDNA encoding nuclear transcription factor MafG from rat VMS (REFERENCES 2).
[3] We got clones encoded a putative transmembrane protein, rhombencephalic expression protein-40 kDa (Rhombex-40) (REFERENCES 3).
[4] We cloned MafG-2, a novel splice variant of MafG, from rat brain by RT-PCR (REFERENCES 4).

Report

(4 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Shimokawa, N., Miura, M.: "Rhombex-29, a novel gene of the PLP/DM2O-M6 family cloned from rat medulla oblongata by differential display"J. Neurosci. Res. 62. 1-8 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shimokawa, N., Okada, J., Miura, M.: "Cloning of MafG homologue from the rat brain by differential display and its expression after hypercapnic stimulation"Mol. Cell. Biochem.. 203. 135-141 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shimokawa, N., Jingu, H., Okada, J., Miura, M.: "Molecular cloning of Rhombex-40 a transmembrane protein from the ventral medullary surface of the rat brain by differential display"Life Sciences. 66. 2183-2191 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shimokawa, N., Kumaki, I., Takayama, K.: "MafG-2 is a novel Maf protein that is expressed by stimulation of extracellular H^+"Cell. Signal.. 13. 835-839 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shimokawa, N., Miura. M.: "Rhombex-29, a novel gene of the PLP/DM20-M6 family cloned from rat medulla oblongata by differential display"J. Neurosci. Res. 62. 1-8 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shimokawa, N., Okada, J., Miura, M.: "Cloning of MafG homologue from the rat brain by differential display and its expression after hypercapnic stimulation"Mol. Cell. Biochem.. 203. 135-141 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shimokawa, N., Jingu, H., Okada, J., Miura, M.: "Molecular cloning of Rhombex-40 a transmembrane protein from the ventral medullary surface of the rat brain by differential display"Life Sciences. 66. 2183-2191 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] Shimokawa, N., Kumaki, I., Takayama, K: "MafG-2 is a novel Maf protein that is expressed by stimulation of extracellular H^+"Cell. Signal. 13. 835-839 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] N.Shimokawa, I.Kumaki, K.Tokoyama: "MafG-2 is a novel Mpf protein that is expressed by stimulation of extracellular H^+"Cellular Signalling. 13. 835-839 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] N.Shimokawa, J.Okada, M.Miura: "Cloning of MafG homologue from the rat brain by differential display and its expression after hypercapnic stimulation"Molecalar and Cellular Biochemistry. 203. 135-141 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Shimokawa, H.Jingu, J.Okada, M.Miura: "Molecular Cloning of Rhombox-40 a transmembrane protein from ventral medullay surface of the rat brain by differential display"Life Sciences. 66. 2183-2191 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] N.Shimokawa, M.Miura: "Rhombex-29, a novel gene of the PLP/OH20-M6 Family cloned from rat medullar of longata by differential display"Journal of Neuroscience Research. 62. 1-8 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Noriaki Shimokawa: "Cloning of Maf G homologue from the rat brain by differential display and its expression after hypercapnic stimulation"Molecular and Cellular Biochemistry. 203. 135-141 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Noriaki Shimokawa: "Molecular cloning of Rhombex-40 a transmembrane protein from the ventral medullary surface of the rot brain by DD"LIFE SCIENCES. (in press). (2000)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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