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Distribution of Glutamate Transporter GLAST in Membranes of Cultured Astrocytes in the Presence of Glutamate Transport Substrates and ATP

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Abstract

Neurotransmitter l-glutamate released at central synapses is taken up and “recycled” by astrocytes using glutamate transporter molecules such as GLAST and GLT. Glutamate transport is essential for prevention of glutamate neurotoxicity, it is a key regulator of neurotransmitter metabolism and may contribute to mechanisms through which neurons and glia communicate with each other. Using immunocytochemistry and image analysis we have found that extracellular d-aspartate (a typical substrate for glutamate transport) can cause redistribution of GLAST from cytoplasm to the cell membrane. The process appears to involve phosphorylation/dephosphorylation and requires intact cytoskeleton. Glutamate transport ligands l -trans-pyrrolidine-2,4-dicarboxylate and dl-threo-3-benzyloxyaspartate but not anti,endo-3,4-methanopyrrolidine dicarboxylate have produced similar redistribution of GLAST. Several representative ligands for glutamate receptors whether of ionotropic or metabotropic type, were found to have no effect. In addition, extracellular ATP induced formation of GLAST clusters in the cell membranes by a process apparently mediated by P2 receptors. The present data suggest that GLAST can rapidly and specifically respond to changes in the cellular environment thus potentially helping to fine-tune the functions of astrocytes.

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Acknowledgments

The project was supported by a grant from the Australian Health Management Fund (VJB) and by a Sydney University Sesquicentennial Grant (VJB).

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Authors and Affiliations

  1. Anatomy and Histology, School of Medical Sciences and Bosch Institute for Biomedical Research, The University of Sydney, Sydney, NSW, 2006, Australia

    Jae-Won Shin, Khoa T. D. Nguyen, Toby Knight & Vladimir J. Balcar

  2. Centre for Clinical Research, Royal Brisbane and Womens Hospital, The University of Queensland, Brisbane, QLD, 4029, Australia

    David V. Pow

  3. Brain and Mind Research Institute, The University of Sydney, Sydney, NSW, 2006, Australia

    Vlado Buljan & Maxwell R. Bennett

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  1. Jae-Won Shin
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  2. Khoa T. D. Nguyen
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  7. Vladimir J. Balcar
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Correspondence to Vladimir J. Balcar.

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The authors J.-W. Shin and K. T. D. Nguyen have contributed equally.

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Shin, JW., Nguyen, K.T.D., Pow, D.V. et al. Distribution of Glutamate Transporter GLAST in Membranes of Cultured Astrocytes in the Presence of Glutamate Transport Substrates and ATP. Neurochem Res 34, 1758–1766 (2009). https://doi.org/10.1007/s11064-009-9982-z

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