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Synaptogenic Assays Using Neurons Cultured on Micropatterned Substrates

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Synapse Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1538))

Abstract

One of the difficulties for studying the mechanisms of synaptogenesis stems from the spatial unpredictability of contact formation between neurons, and the involvement of many parallel adhesive pathways mediating axon/dendrite recognition. To circumvent these limitations, we describe here a method allowing the investigation of synaptic contacts at controlled locations with high precision and statistics. Specifically, primary neurons are cultured on micropatterned substrates comprising arrays of micron-scale dots coated with purified synaptogenic adhesion molecules. Coating the substrates with the homophilic adhesion molecule SynCAM triggers the formation of functional presynaptic structures in axons, while neurexin elicits postsynapses in dendrites from neurons expressing the counter receptor neuroligin. This assay can be combined with various imaging techniques including immunocytochemistry to screen the accumulation of synaptic components, long-term live cell recordings to probe the kinetics of neurite growth and synapse differentiation, as well as high resolution single molecule tracking.

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Acknowledgments

We thank P. Scheiffele and S. Okabe for the generous gift of plasmids; B. Tessier, F. Neca, and Z. Karatas for molecular biology; the Cell culture facility of the Institute; A. Azioune for help on coating protocols, J.B. Sibarita and F. Levet for image analysis; and S. Lafosse for the temporary loan of the Nikon BioStation.

This work received funding from the Centre National de la Recherche Scientifique, Agence Nationale de la Recherche (grants Synapse-2Dt and SynAdh), Conseil Régional Aquitaine and Fondation pour la Recherche Médicale.

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Authors and Affiliations

  1. Interdisciplinary Institute for Neuroscience, University of Bordeaux, UMR 5297, 146 rue Leo Saignat, F-33000, Bordeaux, France

    Katalin Czöndör & Olivier Thoumine

  2. Interdisciplinary Institute for Neuroscience, CNRS, UMR 5297, F-33000, Bordeaux, France

    Katalin Czöndör & Olivier Thoumine

Authors
  1. Katalin Czöndör
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  2. Olivier Thoumine
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Correspondence to Olivier Thoumine .

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Editors and Affiliations

  1. Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA

    Alexandros Poulopoulos

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Czöndör, K., Thoumine, O. (2017). Synaptogenic Assays Using Neurons Cultured on Micropatterned Substrates. In: Poulopoulos, A. (eds) Synapse Development. Methods in Molecular Biology, vol 1538. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6688-2_3

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  • DOI: https://doi.org/10.1007/978-1-4939-6688-2_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6686-8

  • Online ISBN: 978-1-4939-6688-2

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