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Functional Linkage of Neutral amino acid transport system L and voltage-gated Ca^<2+> channels in central nervous systems.

Research Project

Project/Area Number 15590143
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Medical pharmacy
Research InstitutionKyoto Pharmaceutical University

Principal Investigator

FUJITA Takuya  Kyoto Pharmaceutical University, Department of Biochemical Pharmacology, Associate Professor, 薬学部, 助教授 (00247785)

Project Period (FY) 2003 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
KeywordsAmino acids / Transporter / Voltage-gated Ca^<2+> channels / Gabapentin / Astrocytes / Neurons / Nitric oxide / Nitric oxide synthase / 電位依存性Ca^<2+>チャンネル / gabapentin / アストロサイト / ニューロン
Research Abstract

In the present research, I have investigated the involvement of Na^+-independent neutral amino acid transport system L in voltage-gated Ca^<2+> channels blocking action by gabapentin in normal human astrocytes and primary cultured mouse cerebrocortical neurons. In particular, there is no information available for amino acid transport system L in neurons that are thought to be a major target site for gabapentin. Therefore, we primarily investigated the functional characteristics of gabapentin transport into the neurons using primary cultures of mouse cerebrocortical neurons. Cerebrocortical neurons were isolated from 15-day-old-fetal ddY mice. The purity of neurons and contamination of glial cells were confirmed by immunocytochemistry using anti-MAP2 (neuronal marker) and anti-GFAP (glial marker) antibodies, respectively.
The uptake of [^3H]gabapentin in neurons was predominantly Na^+-independent, was stimulated by lowering the extracellular pH, and was inhibited by hydrophobic neutral amino acids. The transport process was saturable with a K_t of 66±0.7 μM for gabapentin uptake. RT PCR revealed that mouse cerebrocortical neurons express the system L amino acid transporters, LAT1 and LAT2, and the heavy chain of the cell surface antigen 4F2 (4F2hc/CD98), which is required for the functional expression of LAT1 and LAT2. Structurally-related compounds of gabapentin, such as 1-amino-1-cyclohexanecarboxylic acid (ACHC), 1-amino-1-cyclopentane- carboxylic acid (cycloleucine), and BCH inhibited the Na^+-independent gabapentin transport in a concentration-dependent manner, and their IC_<50> values were±13, 185±24, and 56±21 μM, respectively. In addition, preloading of various neutral amino acids into neurons markedly enhanced Na^+-independent [^3H]gabapentin uptake (trans-stimulation effect).
These results suggest that functional amino acid transport system L is expressed in mouse cerebrocortical neurons.

Report

(3 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • Research Products

    (26 results)

All 2005 2004 2003 Other

All Journal Article (22 results) Publications (4 results)

  • [Journal Article] Functional expression and adaptive regulation of Na^+-dependent neutral amino acid transporter SNAT2/ATA2 in normal human astrocytes under amino acid starved conditions.2005

    • Author(s)
      Kazunari Tanaka et al.
    • Journal Title

      Neuroscience Letters 378

      Pages: 70-75

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional regulation of Na^+-dependent neutral amino acid transporter ASCT2 by S-nitrosothiols and nitric oxide in Caco-2 cells.2005

    • Author(s)
      Tomomi Uchiyama et al.
    • Journal Title

      FEBS Letters 579

      Pages: 2499-2506

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Transport characteristics of N-acetyl-L-aspartate in rat astrocytes : involvement of sodium-coupled high-affinity carboxylate transporter NaC3/NaDC3-mediated transport system.2005

    • Author(s)
      Takuya Fujita et al.
    • Journal Title

      Journal of Neurochemistry 93

      Pages: 706-714

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional linkage of H^+/peptide transporter PEPT2 and Na^+/H^+ exchanger in primary cultures of astrocytes from mouse cerebral cortex.2005

    • Author(s)
      Miyuki Wada et al.
    • Journal Title

      Brain Research 1044

      Pages: 33-41

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional expression and adaptive regulation of Na^+-dependent neutral amino acid transporter SNAT2/ATA2 in normal human astrocytes under amino acid starved conditions2005

    • Author(s)
      Kazunari Tanaka et al.
    • Journal Title

      Neuroscience Letters 378

      Pages: 70-75

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional regulation of Na^+-dependent neutral amino acid transporter ASCT2 by S-nitrosothiols and nitric oxide in Caco-2 cells2005

    • Author(s)
      Uchiyama Tomomi et al.
    • Journal Title

      FEBS Letters 579

      Pages: 2499-2506

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Transport characteristics of N-acetyl-L-aspartate in rat astrocytes : involvement of sodium-coupled high-affinity carboxylate transporter NaC3/NaDC3-mediated transport system2005

    • Author(s)
      Takuya Fujita et al.
    • Journal Title

      Journal of Neurochemistry 93

      Pages: 706-714

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional linkage of H^+/peptide transporter PEPT2 and Na^+/H^+ exchanger in primary cultures of astrocytes from mouse cerebral cortex2005

    • Author(s)
      Miyuki Wada et al.
    • Journal Title

      Brain Research 1044

      Pages: 33-41

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional expression and adaptive regulation of Na^+-dependent neutral amino acid transporter SNAT1/ATA in normal human astrocytes under amino acid starved condition.2005

    • Author(s)
      Kazunari Tanaka et al.
    • Journal Title

      Neuroscience Letters 378(2)

      Pages: 70-75

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Functional linkage of H^+/peptide transporter PEPT2 and Na^+/H^+ exchanger in primary cultures of astrocytes from mouse cerebral cortex.2005

    • Author(s)
      Miyuki Wada et al.
    • Journal Title

      Brain Research (in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Transport characteristics of N-acetyl-L-aspartate in rat astrocytes : Involvement of sodium-coupled high affinity carboxylate transporter NaC3/NaDC3 mediated transport system.2005

    • Author(s)
      Takuya Fujita et al.
    • Journal Title

      Journal of Neurochemistry 93(in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Functional regulation of Na^+-dependent neutral amino acid transporter ASCT2 by S-nitrosothiols and nitric oxide in Caco-2 cells2005

    • Author(s)
      Tomomi Uchiyama et al.
    • Journal Title

      FEBS Letters (in press)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Functional characterization of brain peptide transporter in rat cerebral cortex : Identification of the high affinity type H^+/peptide transporter PEPT2.2004

    • Author(s)
      Takuya Fujita et al.
    • Journal Title

      Brain Research 997

      Pages: 52-61

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional expression of constitutive nitric oxide synthase regulated by voltage gated Na^+- and Ca^<2+> channels in cultured human astrocytes.2004

    • Author(s)
      Michiko Oka et al.
    • Journal Title

      Glia 46

      Pages: 53-62

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional characterization of brain peptide transporter in rat cerebral cortex : Identification of the high affinity type H+/peptide transporter PEPT22004

    • Author(s)
      Takuya Fujita et al.
    • Journal Title

      Brain Research 997

      Pages: 52-61

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional expression of constitutive nitric oxide synthases regulated by voltage gated Na^+- and Ca ^<2+> channels in cultured human astrocytes2004

    • Author(s)
      Michiko Oka et al.
    • Journal Title

      Glia 46

      Pages: 53-62

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Functional characterization of brain peptide transporter in rat cerebral cortex : identification of the high-affinity type H^+/peptide transporter PEPT2.2004

    • Author(s)
      Takuya Fujita et al.
    • Journal Title

      Brain Research 997(1)

      Pages: 52-61

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Functional expression of constitutive nitric oxide synthases regulated by voltage-gated Na^+- and Ca^<2+>-channels in cultured human astrocytes.2004

    • Author(s)
      Michiko Oka et al.
    • Journal Title

      Glia 46(1)

      Pages: 53-62

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A comparison of Ca^<2+> channel blocking mode between gabapentin and verapamil : Implication for protection against hypoxic injury in rat cerebrocortical slices.2003

    • Author(s)
      Michiko Oka et al.
    • Journal Title

      British Journal of Pharmacology 139

      Pages: 435-443

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Gabapentin blocks L-type and P/Q-type Ca^<2+> channels involved in depolarization-stimulated nitric oxide synthase activity in primary culture of neurons from mouse cerebral cortex.2003

    • Author(s)
      Michiko Oka et al.
    • Journal Title

      Pharmaceutical Research 20

      Pages: 897-899

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] A comparison of Ca^<2+> channel blocking mode between gabapentin and verapamil : Implication for protection against hypoxic injury in rat cerebrocortical slices2003

    • Author(s)
      Michiko Oka et al.
    • Journal Title

      British Journal of Pharmacology 139

      Pages: 435-443

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Gabapentin blocks L-type and P/Q-type Ca^<2+> channels involved in depolarization-stimulated nitric oxide synthase activity in primary culture of neurons from mouse cerebral cortex2003

    • Author(s)
      Michiko Oka et al.
    • Journal Title

      Pharmaceutical Research 20

      Pages: 897-899

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Michiko Oka et al.: "A comparison of Ca^<2+> channel blocking mode between gabapentin and verapamil : implication for protection against hypoxic injury in rat cerebrocortical slices."British Journal of Pharmacology. 139. 435-443 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Michiko Oka et al.: "Gabapentin blocks L-type and P/Q-type Ca^<2+>-channels involved in depolarization-stimulated nitric oxide synthase activity in primary cultures of neurons from mouse cerebral cortex."Pharmaceutical Research. 20. 897-899 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Takuya Fujita et al.: "Functional characterization of brain peptide transporter in rat cerebral cortex : identification of the high affinity type H^+/peptide transporter PEPT2."Brain Research. 997. 52-61 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Michiko Oka et al.: "Functional expression of constitutive nitric oxide synthases regulated by voltage-gated Na^+- and Ca^<2+> channels in cultured human astrocytes."Glia. 46. 53-62 (2004)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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