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A metabonomic study of inhibition of GABA uptake in the cerebral cortex

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Abstract

GABAergic activity is regulated by rapid, high affinity uptake of GABA from the synapse. Perturbation of GABA reuptake has been implicated in neurological disease and inhibitors of GABA transporters (GAT) have been used therapeutically but little detail is known about the ramifications of GAT inhibition on brain neurochemistry. Here, we incubated Guinea pig cortical tissue slices with [3-13C]pyruvate and major, currently available GABA uptake inhibitors. Metabolic fingerprints were generated from these experiments using 13C/1H NMR spectroscopy. These fingerprints were analyzed using multivariate statistical approaches and compared with an existing library of fingerprints of activity at GABA receptors. This approach identified five distinct clusters of metabolic activity induced by blocking GABA uptake. Inhibition of GABA uptake via GAT1 produced patterns similar to activity at mainstream GABAergic synapses in particular those containing α1-subunits but still statistically separable. This indicated that inhibition of GABA uptake, an indirect method of activating GABA receptors, produces different effects to direct receptor activation or to exogenous GABA. The mechanism of inhibitor function also produced different outcomes, with the channel blocker SKF 89976A yielding a unique metabolic response. Blocking GAT1 and GAT3 simultaneously induces a large metabolic response consistent with induction of tonic inhibition via high affinity GABA receptors. Blocking BGT produces patterns similar to activity at less common receptors such as those containing α5 subunits. This approach is useful for determining where in the spectrum of GABAergic responses a particular GABA transport inhibitor is effective.

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Acknowledgements

This work was supported by UNSW (Goldstar award to CR), NewSouth Global and the Australian National Health and Medical Research Council (grant to CR and VJB). The authors are grateful to Ms. Adelle Shasta, and Dr. Donald Thomas of the UNSW Analytical centre for expert technical support.

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Authors and Affiliations

  1. Prince of Wales Medical Research Institute, Brain Sciences UNSW, Barker St, Randwick, NSW, 2031, Australia

    Fatima A. Nasrallah & Caroline Rae

  2. The Bosch Institute and School of Medical Sciences, Sydney Medical School, The University of Sydney, Sydney, NSW, 2006, Australia

    Vladimir J. Balcar

  3. School of Chemistry, The University of New South Wales, Sydney, NSW, 2052, Australia

    Caroline Rae

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  1. Fatima A. Nasrallah
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  2. Vladimir J. Balcar
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  3. Caroline Rae
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Correspondence to Caroline Rae.

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Nasrallah, F.A., Balcar, V.J. & Rae, C. A metabonomic study of inhibition of GABA uptake in the cerebral cortex. Metabolomics 6, 67–77 (2010). https://doi.org/10.1007/s11306-009-0176-0

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