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The Rapid L- and D-Aspartate Uptake in Cultured Astrocytes

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Abstract

Astrocytes in primary culture possess a rapid L-aspartate saturable transport system (Km = 93 μM; Vmax = 81 nmol/min/mg protein), which shows certain stereospecificity since Vmax was 36% lower for D-aspartate uptake. These are values obtained at short incubation time (15 seconds), to obtain approximate initial rate conditions. Metabolic energy inhibitors, rotenone and iodoacetate very potently inhibited the L- and D-aspartate uptake processes, indicating that the transport process is an active one. However, the accumulation of L-aspartate was "enhanced” by inhibitors of L-aspartate metabolism, such as the aspartate aminotransferase inhibitor, aminooxyacetate and L-methionine sulfoximine, an inhibitor of glutamine synthetase, whereas D-aspartate (a non-metabolizable analog of L-aspartate) uptake was not affected. The accumulated levels of L-aspartate in the presence of aminooxyacetate were similar to the levels of D-aspartate. These effects of L-aspartate metabolic inhibitors, suggest that due to metabolism of the the L-aspartate, short incubation time (eg., 15 seconds) is necessary to measure the initial rate of L-aspartate uptake, in order to obtain the "true” kinetic parameters.

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Author notes

  1. Alex S. Bender

    Present address: Department of Pathology, University of Miami School of Medicine, Miami, Florida, 33101

Authors and Affiliations

  1. Department of Pharmacology and Toxicology, College of Pharmacy, University of Utah, Salt Lake City, Utah, 84112

    Alex S. Bender & H. Steve White

  2. Division of Neuropharmacology and Epileptology, Department of Physiology, School of Medicine, University of Utah, Salt Lake City, Utah, 84112

    Dixon M. Woodbury

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  1. Alex S. Bender
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  2. Dixon M. Woodbury
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Bender, A.S., Woodbury, D.M. & White, H.S. The Rapid L- and D-Aspartate Uptake in Cultured Astrocytes. Neurochem Res 22, 721–726 (1997). https://doi.org/10.1023/A:1027358211472

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