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Nanoscale Imaging of Synapses pp 59–74Cite as

Investigating AMPA Receptor Diffusion and Nanoscale Organization at Synapses with High-Density Single-Molecule Tracking Methods

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Part of the book series: Neuromethods ((NM,volume 84))

Abstract

First used in the mid-1980s to complement bulk measurement methods such as fluorescence recovery after photobleaching (FRAP), single-particle tracking techniques have now become a major tool to understand protein mobility and organization in cells. Indeed, bulk measurement methods cannot give access to the full range of heterogeneities resulting from variations across time or from behavior of individual molecules. Only following individual molecules can provide the full distribution of their time-dependent properties. Thanks to improvements in visualization approaches combined with new strategies to label proteins with nanoscale probes, important developments in single-molecule imaging technologies have been made. In this chapter, we will describe the advantages and limitations of the different characteristics of three different approaches used to track single molecules in live neurons: quantum dots (QDs) tracking, sptPALM (single-particle tracking PhotoActivation Light Microscopy) and uPAINT (universal Point Accumulation In Nanoscale Topography). These complementary techniques provide high-density dynamical information revealing localization-specific diffusion properties of molecules on a single cell. We will present the application of these approaches to track AMPA receptors and describe the new insights they give on the dynamic organization of synapses.

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Author information

Authors and Affiliations

  1. Interdisciplinary Institute for Neuroscience (IINS), UMR 5297, University of Bordeaux, Bordeaux, France

    Audrey Constals

  2. Interdisciplinary Institute for Neuroscience (IINS), UMR 5297, CNRS, Bordeaux, France

    Audrey Constals

  3. Interdisciplinary Institute for Neuroscience (IINS), University of Bordeaux, Bordeaux, France

    Eric Hosy & Daniel Choquet

  4. Interdisciplinary Institute for Neuroscience (IINS), CNRS UMR 5297, Bordeaux, France

    Eric Hosy & Daniel Choquet

Authors
  1. Audrey Constals
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  2. Eric Hosy
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  3. Daniel Choquet
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Editor information

Editors and Affiliations

  1. Interdisciplinary Institute for Neuroscience, University of Bordeaux, Bordeaux, France

    U. Valentin Nägerl

  2. Institute of Biology at Ecole Normale Supérieure, Ecole Normale Supérieure, Paris, France

    Antoine Triller

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Constals, A., Hosy, E., Choquet, D. (2014). Investigating AMPA Receptor Diffusion and Nanoscale Organization at Synapses with High-Density Single-Molecule Tracking Methods. In: Nägerl, U., Triller, A. (eds) Nanoscale Imaging of Synapses. Neuromethods, vol 84. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4614-9179-8_4

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  • DOI: https://doi.org/10.1007/978-1-4614-9179-8_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4614-9178-1

  • Online ISBN: 978-1-4614-9179-8

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