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Mechanism of Ca2+/calmodulin-dependent kinase II regulation of AMPA receptor gating

Nature Neuroscience volume 14, pages 727–735 (2011)Cite this article

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Abstract

The function, trafficking and synaptic signaling of AMPA receptors are tightly regulated by phosphorylation. Ca2+/calmodulin-dependent kinase II (CaMKII) phosphorylates the GluA1 AMPA receptor subunit at Ser831 to increase single-channel conductance. We show that CaMKII increases the conductance of native heteromeric AMPA receptors in mouse hippocampal neurons through phosphorylation of Ser831. In addition, co-expression of transmembrane AMPA receptor regulatory proteins (TARPs) with recombinant receptors is required for phospho-Ser831 to increase conductance of heteromeric GluA1-GluA2 receptors. Finally, phosphorylation of Ser831 increases the efficiency with which each subunit can activate, independent of agonist efficacy, thereby increasing the likelihood that more receptor subunits will be simultaneously activated during gating. This underlies the observation that phospho-Ser831 increases the frequency of openings to larger conductances rather than altering unitary conductance. Together, these findings suggest that CaMKII phosphorylation of GluA1-Ser831 decreases the activation energy for an intrasubunit conformational change that regulates the conductance of the receptor when the channel pore opens.

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Figure 1: CaMKII enhances γMEAN of hippocampal AMPA receptors recorded in outside-out patches from cultured hippocampal neurons.
Figure 2: The increase in γMEAN from GluA1-GluA2-stargazin–transfected cells is not caused by a stargazin-induced increase in a subpopulation of homomeric GluA1 receptors.
Figure 3: Phosphorylation of GluA1-Ser831 changes γMEAN independent of agonist efficacy.
Figure 4: GluA1-Ser831 phosphorylation increases the coupling efficiency between agonist binding and gating.
Figure 5: GluA1-S831E phosphomimic mutation increases the frequency of higher conductance openings and increases the single-channel coupling efficiency.

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Acknowledgements

We thank V. Derkach and J. Howe for comments on the manuscript. We thank D. Colquhoun (University College London) for providing software for time course fitting and single channel dwell time and amplitude analysis (SCAN, EKDIST). GluA1 and GluA2 cDNA was provided by P. Seeburg (Max-Planck-Institut für medizinische Forschung) and γ8 cDNA by R. Nicoll (University of California, San Francisco). This work was supported by the US National Institutes of Health (NS068464, NS036654; S.F.T.), the Howard Hughes Medical Institute (R.H.), the Alfred Benzon Foundation (A.S.K., T.G.B.) and the Lundbeck Foundation (A.S.).

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Authors and Affiliations

  1. Department of Pharmacology, Emory University School of Medicine, Atlanta, Georgia, USA

    Anders S Kristensen, Meagan A Jenkins, Tue G Banke & Stephen F Traynelis

  2. Department of Medicinal Chemistry, University of Copenhagen, Copenhagen, Denmark

    Anders S Kristensen

  3. Department of Pharmacology and Pharmacotherapy, University of Copenhagen, Copenhagen, Denmark

    Arne Schousboe

  4. Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Yuichi Makino, Richard C Johnson & Richard Huganir

  5. Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA

    Yuichi Makino, Richard C Johnson & Richard Huganir

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Contributions

A.S.K. and M.A.J. performed patch clamp recordings from recombinant and neuronal receptors; M.A.J. performed all recordings with neurons from transgenic animals. T.G.B. performed a subset of rapid agonist application experiments onto recombinant receptors. Y.M., R.C.J. and R.H. generated knock-in mice for this study. T.G.B. and A.S. performed Xenopus oocyte recordings. S.F.T. assisted with experimental design and participated in data analysis and interpretation. All authors contributed to writing the manuscript.

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Correspondence to Stephen F Traynelis.

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Kristensen, A., Jenkins, M., Banke, T. et al. Mechanism of Ca2+/calmodulin-dependent kinase II regulation of AMPA receptor gating. Nat Neurosci 14, 727–735 (2011). https://doi.org/10.1038/nn.2804

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