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PKA phosphorylation of AMPA receptor subunits controls synaptic trafficking underlying plasticity

Nature Neuroscience volume 6, pages 136–143 (2003)Cite this article

Abstract

The regulated incorporation of AMPA receptors into synapses is important for synaptic plasticity. Here we examine the role of protein kinase A (PKA) in this process. We found that PKA phosphorylation of the AMPA receptor subunits GluR4 and GluR1 directly controlled the synaptic incorporation of AMPA receptors in organotypic slices from rat hippocampus. Activity-driven PKA phosphorylation of GluR4 was necessary and sufficient to relieve a retention interaction and drive receptors into synapses. In contrast, PKA phosphorylation of GluR1 and the activity of calcium/calmodulin-dependent kinase II (CaMKII) were both necessary for receptor incorporation. Thus, PKAphosphorylation of AMPA receptor subunits contributes to diverse mechanisms underlying synaptic plasticity.

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Figure 1: PKA activity is necessary and sufficient for GluR4–GFP incorporation into synapses.
Figure 2: GluR4–GFP is driven into synapses rapidly upon acute PKA activation.
Figure 3: Regulated phosphorylation of GluR4 and GluR1 by PKA and CaMKII.
Figure 4: GluR4 Ser842 controls a retention interaction.
Figure 5: PKA activity is necessary but not sufficient for the CaMKII-driven incorporation of GluR1 into synapses.
Figure 6: Phosphorylation of PKA target S845, but not CaMKII target S831, is required for stable LTP.

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Acknowledgements

We thank N. Dawkins-Pisani and J. Kuchibhotia for technical assistance. This work was supported by National Alliance for Research in Schizophrenia and Depression (NARSAD to J.E.), National Institutes of Health NS32827 (to R.M.), Alle Davis and Maxine Harrison Endowment (to R.M.), The Howard Hughes Medical Institute (to R.L.H.) and National Institutes of Health NS36715 (to R.L.H.).

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Author notes

  1. José A. Esteban

    Present address: Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan, 48109, USA

Authors and Affiliations

  1. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 11724, USA

    José A. Esteban, Song-Hai Shi, Christopher Wilson & Roberto Malinow

  2. Department of Neuroscience, Howard Hughes Medical Institute, The Johns Hopkins University School of Medicine, Baltimore, 21205, Maryland, USA

    Mutsuo Nuriya & Richard L. Huganir

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Correspondence to José A. Esteban or Roberto Malinow.

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Supplementary information

Supplementary Fig. 1.

Specificity of GluR4 phospho-specific antibody. (a) Rat cerebellum P2 fraction was prepared and resolved by SDS-PAGE. Membranes were treated with l phosphatase (1,2000 U/ml) or control solution and then blocked with each peptide (1mg/ml) as indicated. Membranes were then probed with anti GluR4 phospho-S842 specific antibody and reprobed with anti C-terminus antibody. (b) GluR4 full length constructs (wild type or S842A mutant) were transfected into HEK293T cells. After ~48hrs, cells were treated with forskolin (20 μM, 15min at 37 ºC) or vehicle (DMSO). Cell lysates were prepared and tested with anti-GluR4 C-terminus antibody and phospho-S842 specific antibody by western blotting. (JPG 33 kb)

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Esteban, J., Shi, SH., Wilson, C. et al. PKA phosphorylation of AMPA receptor subunits controls synaptic trafficking underlying plasticity. Nat Neurosci 6, 136–143 (2003). https://doi.org/10.1038/nn997

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