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Distinct molecular mechanisms and divergent endocytotic pathways of AMPA receptor internalization

Nature Neuroscience volume 3, pages 1282–1290 (2000)Cite this article

Abstract

Internalization of postsynaptic AMPA receptors depresses excitatory transmission, but the underlying dynamics and mechanisms of this process are unclear. Using immunofluorescence and surface biotinylation, we characterized and quantified basal and regulated AMPA receptor endocytosis in cultured hippocampal neurons, in response to synaptic activity, AMPA and insulin. AMPA-induced AMPA receptor internalization is mediated in part by secondary activation of voltage-dependent calcium channels, and in part by ligand binding independent of receptor activation. Although both require dynamin, insulin- and AMPA-induced AMPA receptor internalization are differentially dependent on protein phosphatases and sequence determinants within the cytoplasmic tails of GluR1 and GluR2 subunits. AMPA receptors internalized in response to AMPA stimulation enter a recycling endosome system, whereas those internalized in response to insulin diverge into a distinct compartment. Thus, the molecular mechanisms and intracellular sorting of AMPA receptors are diverse, and depend on the internalizing stimulus.

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Figure 1: Rapid ligand-dependent endocytosis of AMPA receptors in cultured hippocampal neurons.
Figure 5: Differential inhibition of AMPA- and insulin-induced AMPA receptor endocytosis by inhibitors of protein phosphatases and L-type calcium channels.
Figure 2: Time course of basal, AMPA-, and insulin-induced internalization of AMPA receptors.
Figure 3: Continued AMPA receptor internalization during the plateau phase of internalized AMPA receptor signal.
Figure 4: AMPA- and insulin-induced AMPA receptor internalization through a clathrin-mediated, dynamin-dependent mechanism.
Figure 6: Differential sorting of internalized AMPA receptors after AMPA and insulin stimulation.
Figure 7: Differential subunit and sequence requirements for AMPA- and insulin-induced AMPA receptor endocytosis.

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Acknowledgements

We thank R.B. Vallee (Worcester Institute, Shrewsbury, Massachusetts) for dynamin I cDNAs, R. Scheller and R. Prekeris (Stanford University School of Medicine, Stanford, California) for syntaxin 13 antibodies, and R. Huganir (Johns Hopkins University, Baltimore, Maryland) for GluR1 antibodies. M.S. is Assistant Investigator of the Howard Hughes Medical Institute. Y.T.W. is a Research Scholar of the Heart and Stroke Foundation of Canada. Supported by grants from US National Institute of Health (NS35050) to M.S. and Heart and Stroke Foundation of Ontario (NA-3762), the Medical Research Council of Canada, and the EJLB Foundation to Y.T.W. S.H.L. was supported in part by a postdoctoral fellowship from KOSEF.

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  1. Howard Hughes Medical Institute, Massachusetts General Hospital (Wellman 423), 50 Blossom Street, Boston, 02114, Massachusetts, USA

    Jerry W. Lin, Kelly Foster, Sang Hyoung Lee, Michael Wyszynski & Morgan Sheng

  2. Department of Neurobiology, Harvard Medical School, 220 Longwood Ave., Boston, 02115, Massachusetts, USA

    Jerry W. Lin, Kelly Foster, Sang Hyoung Lee, Michael Wyszynski & Morgan Sheng

  3. Programme in Brain and Behavior and Division of Pathology, Hospital for Sick Children, 555 University Ave., Toronto, M5G 1X8, Ontario, Canada

    William Ju, Gholamreza Ahmadian & Yu Tian Wang

  4. Department of Laboratory Medicine and Pathobiology, University of Toronto, 100 College St., Toronto, M5G 1X8, Ontario, Canada

    William Ju, Gholamreza Ahmadian & Yu Tian Wang

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Lin, J., Ju, W., Foster, K. et al. Distinct molecular mechanisms and divergent endocytotic pathways of AMPA receptor internalization. Nat Neurosci 3, 1282–1290 (2000). https://doi.org/10.1038/81814

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