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

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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 x_C^-, 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 x_C^-, and GLAST were expressed in TR-AST, which has the characteristics of type 2 astrocytes and is able to transport acidic amino acids.