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Subunit-specific temporal and spatial patterns of AMPA receptor exocytosis in hippocampal neurons

Nature Neuroscience volume 4, pages 917–926 (2001)Cite this article

Abstract

Using a thrombin cleavage assay in cultured hippocampal neurons, we studied the kinetics, regulation and site of AMPA receptor surface delivery. Surface insertion of the GluR1 subunit occurs slowly in basal conditions and is stimulated by NMDA receptor activation and insulin, whereas GluR2 exocytosis is constitutively rapid. Although both subunits ultimately concentrate in synapses, GluR1 and GluR2 show different spatial patterns of surface accumulation, consistent with GluR1 being inserted initially at extrasynaptic sites and GluR2 being inserted more directly at synapses. The spatiotemporal pattern of surface accumulation is determined by the cytoplasmic tails of GluR subunits, and in heteromeric receptors, GluR1 acts dominantly over GluR2. We propose that GluR1 controls the exocytosis and GluR2/3, the recycling and endocytosis of AMPA receptors.

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Figure 1: Specificity of thrombin cleavage of surface HA/T-GluR.
Figure 2: Time course of surface delivery of HA/T-GluR1 and -GluR2 in cultured hippocampal neurons.
Figure 3: Time course of HA/T-GluR1 and HA/T-GluR2 recycling in hippocampal neurons.
Figure 4: Time course of exocytosis of GluR chimeras and heteromeric GluR1/GluR2 receptors.
Figure 5: Regulation of exocytosis of HA/T-GluR1 and -GluR2 by NMDA receptor activation and insulin.
Figure 6: Effect of C-terminal mutations on the surface level and exocytosis of GluR1.
Figure 7: Effect of C-terminal mutations on surface levels and exocytosis of GluR2.
Figure 8: Synaptic concentration of HA/T-GluR1 and HA/T-GluR2 in hippocampal neurons.
Figure 9: Differential spatiotemporal pattern of surface accumulation of HA/T-GluR1 and -GluR2.

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Acknowledgements

We thank C. Sala, S. Hyoung Lee, M. Wyszynski and Y. Tian Wang for discussions. This work was supported by NIH grant NS 35050 (M.S.). M.S. is Associate Investigator of the Howard Hughes Medical Institute.

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Author notes

  1. Morgan Sheng

    Present address: Center for Learning and Memory, Howard Hughes Medical Institute, Massachusetts Institute of Technology (E18-215), 77 Massachusetts Avenue, Cambridge, Massachusetts, 02139, USA

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  1. Department of Neurobiology and Howard Hughes Medical Institute, Massachusetts General Hospital and Harvard Medical School, 50 Blossom Street, Boston, 02114, Massachusetts, USA

    Maria Passafaro, Valentin Piëch & Morgan Sheng

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Correspondence to Morgan Sheng.

Supplementary information

Supplementary Fig.1.

A model for postsynaptic AMPA receptor trafficking.AMPA receptors are represented as heteromers of GluR1/GluR2 or GluR2/GluR3.GluR1 controls the regulated exocytosis of AMPA receptors (acting dominantly in GluR1/GluR2 heteromers).GluR2 mediates the constitutive recycling and regulated endocytosis of AMPA receptors.(See text for details). (GIF 15 kb)

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Passafaro, M., Piëch, V. & Sheng, M. Subunit-specific temporal and spatial patterns of AMPA receptor exocytosis in hippocampal neurons. Nat Neurosci 4, 917–926 (2001). https://doi.org/10.1038/nn0901-917

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