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Differential activity-dependent regulation of the lateral mobilities of AMPA and NMDA receptors

Nature Neuroscience volume 7, pages 695–696 (2004)Cite this article

Abstract

The basis for differences in activity-dependent trafficking of AMPA receptors (AMPARs) and NMDA receptors (NMDARs) remains unclear. Using single-molecule tracking, we found different lateral mobilities for AMPARs and NMDARs: changes in neuronal activity modified AMPAR but not NMDAR mobility, whereas protein kinase C activation modified both. Differences in mobility were mainly detected for extrasynaptic AMPARs, suggesting that receptor diffusion between synaptic and extrasynaptic domains is involved in plasticity processes.

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Figure 1: Differential lateral diffusion of AMPARs (░) and NMDARs (▪) at the surface of 10-d.i.v. hippocampal neurons.
Figure 2: Neuronal activity differentially regulates AMPAR and NMDAR lateral diffusions.

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Acknowledgements

We thank D. Bouchet for hippocampal neuron cultures. This work was supported by grants from Conseil Régional d'Aquitaine, the European Community (QLG3-CT-2001-02089) and the BBSRC (UK).

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Authors and Affiliations

  1. UMR 5091 CNRS–Université Bordeaux 2, Bordeaux, 33077, France

    Laurent Groc, Martin Heine & Daniel Choquet

  2. CNRS-UMR 5798, Talence, France

    Laurent Cognet & Brahim Lounis

  3. School of Pharmacy, University of London, London, UK

    Kieran Brickley & F Anne Stephenson

Authors
  1. Laurent Groc
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  2. Martin Heine
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  7. Daniel Choquet
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Correspondence to Daniel Choquet.

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Supplementary information

Supplementary Fig. 1

Co-localization of Deep Red Mitotrack and synaptotagmin clusters on live neurons. (a-b) Staining of Deep Red Mitotrack and synaptotagmin (polyclonal rabbit anti-synaptotagmin, a gift from C. Dotti coupled to an anti-Fab Zenon 488) on 10 DIV dendritic branches. Neurons were incubated with Deep Red Mitotrack for 1 min (1nM) and then depolarized with KCl (40 mM, 1-2 min) to load synaptotagmin antibody within synaptic vesicles Note the punctuate staining. (c) Merged images from (a) and (b) show that Deep Red Mitotrack and synaptotagmin clusters do colocalize. (d) The quantification of the Mitotrack and synaptotagmin cluster colocalization was done using Metamorph (bar graph: mean ± s.e.m., n = 6 neurons). M = Mitotrack ; S = synaptotagmin. (PDF 38 kb)

Supplementary Fig. 2

Comparison between Cy3-single molecule and quantum dot (QD)-single trajectory for the GluR2 subunit in extrasynaptic and synaptic membranes of 8-15 DIV rat hippocampal neurons. GluR2 Cy3-single molecules were tracked and analyzed as described (article and Supplementary Methods). The QD 605 (emission) conjugated with protein A (Quantum Dot Corporation, Hayward, USA) were visualized by an intensified CCD camera system (Pentamax, Princeton Instruments, Trenton, USA) after excitation with an Argon laser (488 nm excitation). Neurons were first incubated with anti-GluR2 antibodies for 10-15 min at 37°C, incubated with the protein A-conjugated QDs (1 min), and single QDs were tracked for the following 30 min at 30°C (each QT trajectory is at least 100 s). For the Cy3-single molecule experiments, synapses were labelled with the Deep Red Mitotrack (see Supplementary Methods). (a) In the extrasynaptic membrane, GluR2 diffusion distribution from Cy3-single molecule (median = 0.01 μm2/s, inter-quartile range (IQR) 25%-75% = 0.00015-0.09, n = 1722) or single QD (median = 0.044 μm2/s, IQR = 0.00031-0.15, n = 116) was similar (Mann-Whitney test, P = 0.34). (b) Within synapses, diffusion distributions were however significantly different (P < 0.0001) since Cy3-single molecule (median = 0.028 μm2/s, IQR = 0.0019-0.12, n = 154) diffusion was 5-times faster than the single QD (median = 0.005 μm2/s, IQR = 0.0005-0.03, n = 59). Such slower diffusion of QDs may be due to the size of the synaptic cleft (approx. 10-15 nm) or to the high density of receptor within the cleft resulting in cross-linking of the multi-conjugated QDs. (PDF 10 kb)

Supplementary Methods (PDF 125 kb)

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Groc, L., Heine, M., Cognet, L. et al. Differential activity-dependent regulation of the lateral mobilities of AMPA and NMDA receptors. Nat Neurosci 7, 695–696 (2004). https://doi.org/10.1038/nn1270

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