Synaptogenic Assays Using Neurons Cultured on Micropatterned Substrates.

1. Methods Mol Biol. 2017;1538:29-44.

Synaptogenic Assays Using Neurons Cultured on Micropatterned Substrates.

Czöndör K(1)(2), Thoumine O(3)(4).

Author information:
(1)Interdisciplinary Institute for Neuroscience, University of Bordeaux, UMR
5297, 146 rue Leo Saignat, F-33000, Bordeaux, France.
(2)Interdisciplinary Institute for Neuroscience, CNRS, UMR 5297, F-33000,
Bordeaux, France.
(3)Interdisciplinary Institute for Neuroscience, University of Bordeaux, UMR
5297, 146 rue Leo Saignat, F-33000, Bordeaux, France.
.
(4)Interdisciplinary Institute for Neuroscience, CNRS, UMR 5297, F-33000,
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
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.

DOI: 10.1007/978-1-4939-6688-2_3
PMID: 27943181 [Indexed for MEDLINE]

Auteurs Bordeaux Neurocampus