Synaptogenic Assays Using Primary Neurons Cultured on Micropatterned Substrates

Katalin Czöndör, Nathalie Piette, Béatrice Tessier, Vincent Studer, Olivier Thoumine
Methods in Molecular Biology. 2025-01-01; : 3-26
DOI: 10.1007/978-1-0716-4446-1_1

Czöndör K(1), Piette N(1)(2), Tessier B(1), Studer V(1), Thoumine O(3).

Author information:
(1)University of Bordeaux, CNRS UMR 5297, Interdisciplinary Institute for
Neuroscience, IINS, Bordeaux, France.
(2)Alveole S.A., Paris, France.
(3)University of Bordeaux, CNRS UMR 5297, Interdisciplinary Institute for
Neuroscience, IINS, Bordeaux, France. .

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
for the investigation of biomimetic 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 SynCAM1 triggers the formation of functional
presynaptic structures in axons, while neurexin-1β elicits postsynapses in
dendrites from neurons expressing the counter receptor neuroligin-1. This assay
can be combined with various optical 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.