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NMDA receptor activity downregulates KCC2 resulting in depolarizing GABAA receptor–mediated currents

Nature Neuroscience volume 14, pages 736–743 (2011)Cite this article

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Abstract

KCC2 is a neuron-specific K+-Cl− co-transporter that maintains a low intracellular Cl− concentration that is essential for hyperpolarizing inhibition mediated by GABAA receptors. Deficits in KCC2 activity occur in disease states associated with pathophysiological glutamate release. However, the mechanisms by which elevated glutamate alters KCC2 function are unknown. The phosphorylation of KCC2 residue Ser940 is known to regulate its surface activity. We found that NMDA receptor activity and Ca2+ influx caused the dephosphorylation of Ser940 in dissociated rat neurons, leading to a loss of KCC2 function that lasted longer than 20 min. Protein phosphatase 1 mediated the dephosphorylation events of Ser940 that coincided with a deficit in hyperpolarizing GABAergic inhibition resulting from the loss of KCC2 activity. Blocking dephosphorylation of Ser940 reduced the glutamate-induced downregulation of KCC2 and substantially improved the maintenance of hyperpolarizing GABAergic inhibition. Reducing the downregulation of KCC2 therefore has therapeutic potential in the treatment of neurological disorders.

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Figure 1: The furosemide-sensitive K+-Cl− pump KCC2 generates hyperpolarizing GABA-activated currents.
Figure 2: Glutamate application switches GABA transmission to depolarizing and excitatory.
Figure 3: Glutamate shifts EGABA to more depolarized potentials.
Figure 4: Glutamate induces the dephosphorylation of Ser940 and the degradation of KCC2.
Figure 5: Glutamate mediates its effects on KCC2 via a Ca2+-dependent mechanism.
Figure 6: The glutamate-induced effects on KCC2 were mediated by NMDA receptors.
Figure 7: The dephosphorylation of Ser940 and internalization of KCC2 caused by glutamate is dependent on PP1.
Figure 8: OKA reduced the glutamate-induced shift in EGABA.

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Acknowledgements

We thank Y. Haydon and L.K. Chau for critical comments on the manuscript and H. Tang and L. Silayeva for providing technical assistance. The work was supported in part by US National Institutes of Health/National Institute of Neurological Disorders and Stroke grants NS036296, NS047478, NS048045 and NS054900. H.H.C.L. is supported by a Postdoctoral Fellowship from the American Heart Association.

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Author notes

  1. Henry H C Lee

    Present address: Present address: Department of Neurology, F.M. Kirby Neurobiology Center, Children's Hospital Boston, Harvard Medical School, Boston, Massachusetts, USA.,

  2. Henry H C Lee and Tarek Z Deeb: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Neuroscience, Tufts University, Boston, Massachusetts, USA

    Henry H C Lee, Tarek Z Deeb, Joshua A Walker, Paul A Davies & Stephen J Moss

  2. Department of Neuroscience, Physiology and Pharmacology, University College, London, UK

    Stephen J Moss

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Contributions

H.H.C.L. and J.A.W. performed biochemical experiments, and T.Z.D. and P.A.D. performed electrophysiological recordings. H.H.C.L., T.Z.D. and S.J.M. designed the experiments and wrote the manuscript.

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Correspondence to Stephen J Moss.

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Lee, H., Deeb, T., Walker, J. et al. NMDA receptor activity downregulates KCC2 resulting in depolarizing GABAA receptor–mediated currents. Nat Neurosci 14, 736–743 (2011). https://doi.org/10.1038/nn.2806

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