Direct imaging of lateral movements of AMPA receptors inside synapses

Trafficking of AMPA receptors in and out of synapses is crucial for synaptic plasticity. Previous studies have focused on the role of endo/exocytosis processes or that of lateral diffusion of extra‐synaptic receptors. We have now directly imaged AMPAR movements inside and outside synapses of live neurons using single‐ molecule fluorescence microscopy. Inside individual synapses, we found immobile and mobile receptors, which display restricted diffusion. Extra‐synaptic receptors display free diffusion. Receptors could also exchange between these membrane compartments through lateral diffusion. Glutamate application increased both receptor mobility inside synapses and the fraction of mobile receptors present in a juxtasynaptic region. Block of inhibitory transmission to favor excitatory synaptic activity induced a transient increase in the fraction of mobile receptors and a decrease in the proportion of juxtasynaptic receptors. Altogether, our data show that rapid exchange of receptors between a synaptic and extra‐synaptic localization occurs through regulation of receptor diffusion inside synapses.

[1]  M. Sheng,et al.  Postsynaptic Signaling and Plasticity Mechanisms , 2002, Science.

[2]  M. Ehlers,et al.  Dynamics and Regulation of Clathrin Coats at Specialized Endocytic Zones of Dendrites and Spines , 2002, Neuron.

[3]  R. Nicoll,et al.  Direct interactions between PSD-95 and stargazin control synaptic AMPA receptor number , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  D. Choquet,et al.  Regulation of AMPA receptor lateral movements , 2002, Nature.

[5]  E. Ziff,et al.  Receptor trafficking and the plasticity of excitatory synapses , 2002, Current Opinion in Neurobiology.

[6]  A. Sergé,et al.  Receptor Activation and Homer Differentially Control the Lateral Mobility of Metabotropic Glutamate Receptor 5 in the Neuronal Membrane , 2002, The Journal of Neuroscience.

[7]  W. Webb,et al.  Precise nanometer localization analysis for individual fluorescent probes. , 2002, Biophysical journal.

[8]  G. Westbrook,et al.  Mobile NMDA Receptors at Hippocampal Synapses , 2002, Neuron.

[9]  M. Hallek,et al.  Real-Time Single-Molecule Imaging of the Infection Pathway of an Adeno-Associated Virus , 2001, Science.

[10]  T. Schikorski,et al.  Inactivity Produces Increases in Neurotransmitter Release and Synapse Size , 2001, Neuron.

[11]  Mark F. Bear,et al.  Internalization of ionotropic glutamate receptors in response to mGluR activation , 2001, Nature Neuroscience.

[12]  V. Piëch,et al.  Subunit-specific temporal and spatial patterns of AMPA receptor exocytosis in hippocampal neurons , 2001, Nature Neuroscience.

[13]  T. Yanagida,et al.  Single-Molecule Analysis of Chemotactic Signaling in Dictyostelium Cells , 2001, Science.

[14]  Mark von Zastrow,et al.  Role of ampa receptor endocytosis in synaptic plasticity , 2001, Nature Reviews Neuroscience.

[15]  A. Triller,et al.  Fast and reversible trapping of surface glycine receptors by gephyrin , 2001, Nature Neuroscience.

[16]  Wei-Yang Lu,et al.  Activation of Synaptic NMDA Receptors Induces Membrane Insertion of New AMPA Receptors and LTP in Cultured Hippocampal Neurons , 2001, Neuron.

[17]  R. Nicoll,et al.  Contribution of cytoskeleton to the internalization of AMPA receptors. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Dane M. Chetkovich,et al.  Stargazin regulates synaptic targeting of AMPA receptors by two distinct mechanisms , 2000, Nature.

[19]  Mark von Zastrow,et al.  Regulation of AMPA receptor endocytosis by a signaling mechanism shared with LTD , 2000, Nature Neuroscience.

[20]  M. Sheng,et al.  Distinct molecular mechanisms and divergent endocytotic pathways of AMPA receptor internalization , 2000, Nature Neuroscience.

[21]  Richard L. Huganir,et al.  Postsynaptic organisation and regulation of excitatory synapses , 2000, Nature Reviews Neuroscience.

[22]  M. Ehlers,et al.  Reinsertion or Degradation of AMPA Receptors Determined by Activity-Dependent Endocytic Sorting , 2000, Neuron.

[23]  E. Gouaux,et al.  Mechanisms for Activation and Antagonism of an AMPA-Sensitive Glutamate Receptor Crystal Structures of the GluR2 Ligand Binding Core , 2000, Neuron.

[24]  Z. Nusser.  AMPA amd NMDA receptors: similarities and differences in their synaptic distribution , 2000, Current Opinion in Neurobiology.

[25]  J. Henley,et al.  Interactions between AMPA receptors and intracellular proteins , 2000, Neuropharmacology.

[26]  D. Linden,et al.  Expression of Cerebellar Long-Term Depression Requires Postsynaptic Clathrin-Mediated Endocytosis , 2000, Neuron.

[27]  Yu Tian Wang,et al.  Regulation of AMPA Receptor–Mediated Synaptic Transmission by Clathrin-Dependent Receptor Internalization , 2000, Neuron.

[28]  Gerald Kada,et al.  Properties of lipid microdomains in a muscle cell membrane visualized by single molecule microscopy , 2000, The EMBO journal.

[29]  R. Nicoll,et al.  Dynamin-dependent endocytosis of ionotropic glutamate receptors. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Mark von Zastrow,et al.  Role of AMPA Receptor Cycling in Synaptic Transmission and Plasticity , 1999, Neuron.

[31]  Petter Laake,et al.  Different modes of expression of AMPA and NMDA receptors in hippocampal synapses , 1999, Nature Neuroscience.

[32]  G. Collingridge,et al.  Surface Expression of AMPA Receptors in Hippocampal Neurons Is Regulated by an NSF-Dependent Mechanism , 1999, Neuron.

[33]  Mark von Zastrow,et al.  Rapid redistribution of glutamate receptors contributes to long-term depression in hippocampal cultures , 1999, Nature Neuroscience.

[34]  W. Moerner,et al.  Illuminating single molecules in condensed matter. , 1999, Science.

[35]  R. Dingledine,et al.  The glutamate receptor ion channels. , 1999, Pharmacological reviews.

[36]  R. Nicoll,et al.  Rapid, Activation-Induced Redistribution of Ionotropic Glutamate Receptors in Cultured Hippocampal Neurons , 1999, The Journal of Neuroscience.

[37]  T. Schikorski,et al.  Quantitative Ultrastructural Analysis of Hippocampal Excitatory Synapses Materials and Methods Terminology Fixation and Embedding , 2022 .

[38]  H Schindler,et al.  Single-molecule microscopy on model membranes reveals anomalous diffusion. , 1997, Biophysical journal.

[39]  W. Moerner,et al.  Three-Dimensional Imaging of Single Molecules Solvated in Pores of Poly(acrylamide) Gels , 1996, Science.

[40]  Werner Baumgartner,et al.  Characterization of Photophysics and Mobility of Single Molecules in a Fluid Lipid Membrane , 1995 .

[41]  A. Kusumi,et al.  Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cells. , 1993, Biophysical journal.

[42]  R. Cherry,et al.  Tracking of cell surface receptors by fluorescence digital imaging microscopy using a charge-coupled device camera. Low-density lipoprotein and influenza virus receptor mobility at 4 degrees C. , 1992, Journal of cell science.

[43]  R. Malenka,et al.  AMPA receptor trafficking and synaptic plasticity. , 2002, Annual review of neuroscience.

[44]  Michel Orrit,et al.  Fluorescence Spectroscopy of Single Biomolecules Shimon Weiss , 1999 .

[45]  K. Jacobson,et al.  Structural mosaicism on the submicron scale in the plasma membrane. , 1998, Biophysical journal.