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Evaluation of Drugs Acting at Glutamate Transporters in Organotypic Hippocampal Cultures: New Evidence on Substrates and Blockers in Excitotoxicity

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Abstract

Removal of L-glutamate (Glu) from the synapse is critical to maintain normal transmission and to prevent excitotoxicity, and is performed exclusively by excitatory amino acid transporters (EAATs). We investigated the effects of substrates and blockers of EAATs on extracellular Glu and cellular viability in organotypic cultures of rat hippocampus. Seven-day treatment with a range of drugs (L-trans-pyrrolidine-2,4-dicarboxylate, (2S,4R)-4-methyl-glutamate, (±)-threo-3-methylglutamate and DL-threo-β-benzyloxyaspartate), in the presence of 300 μM added Glu, resulted in increased extracellular Glu and a significant correlation between Glu concentration and cellular injury (as indicated by lactate dehydrogenase release). In contrast, (2S,3S,4R)-2-(carboxycyclopropyl)glycine (L-CCG-III) exerted a novel neuroprotection against this toxicity, and elevations in extracellular Glu were not toxic in the presence of this compound. Similar results were obtained following two-week treatment of cultures without added Glu. Whilst blockade of GLT-1 alone was relatively ineffective in producing excitotoxic injury, heteroexchange of Glu by EAAT substrates may exacerbate excitotoxicity.

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Authors and Affiliations

  1. Department of Pharmacology, Monash University, Australia

    Ross D. O'Shea, Melissa V. Fodera & Karina Apricó

  2. Department of Physiology, University of Oslo Institute of Basic Medical Sciences, Oslo, Norway

    Yvette Dehnes & Niels C. Danbolt

  3. Tocris Cookson Ltd., Bristol, United Kingdom

    Duncan Crawford

  4. Department of Pharmacology, Monash University, Australia

    Philip M. Beart

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  1. Ross D. O'Shea
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  2. Melissa V. Fodera
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  5. Niels C. Danbolt
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O'Shea, R.D., Fodera, M.V., Apricó, K. et al. Evaluation of Drugs Acting at Glutamate Transporters in Organotypic Hippocampal Cultures: New Evidence on Substrates and Blockers in Excitotoxicity. Neurochem Res 27, 5–13 (2002). https://doi.org/10.1023/A:1014813518604

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