Acidic Amino Acid Transport Characteristics of a Newly Developed Conditionally Immortalized Rat Type 2 Astrocyte Cell Line (TR-AST).

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  • Tetsuka Kazuhiro
    Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Tohoku University
  • Hosoya Ken-ichi
    Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University CREST of Japan Science and Technology Corporation (JST)
  • Ohtsuki Sumio
    Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Tohoku University CREST of Japan Science and Technology Corporation (JST) New Industry Creation Hatchery Center, Tohoku University
  • Takanaga Hitomi
    Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Tohoku University CREST of Japan Science and Technology Corporation (JST) New Industry Creation Hatchery Center, Tohoku University
  • Yanai Nobuaki
    Department of Cell Biology, Institute of Development, Aging and Cancer, Tohoku University
  • Ueda Masatsugu
    YS New Technology Institute Inc.
  • Obinata Masuo
    CREST of Japan Science and Technology Corporation (JST) Department of Cell Biology, Institute of Development, Aging and Cancer, Tohoku University
  • Terasaki Tetsuya
    Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Tohoku University CREST of Japan Science and Technology Corporation (JST) New Industry Creation Hatchery Center, Tohoku University

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Abstract

To characterize acidic amino acid transport in type 2 astrocytes, we established conditionally immortalized rat astrocyte cell lines (TR-AST) from newly developed transgenic rats harboring temperature-sensitive SV40 large T-antigen gene. TR-AST exhibited positive immunostaining for anti-GFAP antibody and A2B5 antibody, characteristics associated with type 2 astrocytes, and expressed glutamine synthetase. Acidic amino acid transporters, GLT-1 and system xC-, which consists of xCT and 4F2hc, were expressed in all TR-ASTs by RT-PCR. On the other hand, GLAST expression was found in TR-AST3 and 5. The characteristics of [3H]L-glutamic acid (L-Glu) uptake by TR-AST5 include an Na+-dependent and Na+-independent manner, concentration-dependence, and inhibition by L-aspartic acid (L-Asp) and D-aspartic acid (D-Asp). The corresponding Michaelis-Menten constants for the Na+-dependent and Na+-independent process were 36.3 μM and 155 μM, respectively. [3H]L-Asp and [3H]D-Asp uptake by TR-AST5 had an Na+-dependent and Na+-independent manner. This study demonstrated that GLT-1, system xC-, and GLAST were expressed in TR-AST, which has the characteristics of type 2 astrocytes and is able to transport acidic amino acids.

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