title: Synaptogenic Assays Using Neurons Cultured on Micropatterned Substrates.
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.
authors.affiliation: Interdisciplinary Institute for Neuroscience, University of Bordeaux, UMR 5297, 146 rue Leo Saignat, F-33000, Bordeaux, France.|Interdisciplinary Institute for Neuroscience, CNRS, UMR 5297, F-33000, Bordeaux, France.
authors.forename: Katalin
authors.initials: K
authors.lastname: Czöndör
authors.affiliation: Interdisciplinary Institute for Neuroscience, University of Bordeaux, UMR 5297, 146 rue Leo Saignat, F-33000, Bordeaux, France. olivier.thoumine@u-bordeaux.fr.|Interdisciplinary Institute for Neuroscience, CNRS, UMR 5297, F-33000, Bordeaux, France. olivier.thoumine@u-bordeaux.fr.
authors.forename: Olivier
authors.initials: O
authors.lastname: Thoumine
mesh_terms: D000818:Animals; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D055808:Drug Discovery / Q000379:methods*; D005455:Fluorescent Antibody Technique; D057054:Molecular Imaging / Q000379:methods; D009474:Neurons / Q000187:drug effects* / Q000502:physiology*; D017729:Presynaptic Terminals / Q000187:drug effects / Q000378:metabolism; D017966:Pyramidal Cells / Q000187:drug effects / Q000502:physiology; D051381:Rats; D018091:Receptors, AMPA / Q000378:metabolism; D013569:Synapses / Q000187:drug effects* / Q000502:physiology*
chemical_list: D015816:Cell Adhesion Molecules, Neuronal; D018091:Receptors, AMPA
keywords: Adhesion molecules; Micropatterned substrates; N-cadherin; Neurexin; Neuroligin; SynCAM; Synapse formation