Micropatterned substrates coated with neuronal adhesion molecules for high-content study of synapse formation

Abstract

Studying the roles of different proteins and the mechanisms involved in synaptogenesis is hindered by the complexity and heterogeneity of synapse types, and by the spatial and temporal unpredictability of spontaneous synapse formation. Here we demonstrate a robust and high-content method to induce selectively presynaptic or postsynaptic structures at controlled locations. Neurons are cultured on micropatterned substrates comprising arrays of micron-scale dots coated with various synaptogenic adhesion molecules. When plated on neurexin-1b-coated micropatterns, neurons expressing neuroligin-1 exhibit specific dendritic organization and selective recruitment of the postsynaptic scaffolding molecule PSD-95. Furthermore, functional AMPA receptors are trapped at neurexin-1b dots, as revealed by live-imaging experiments. In contrast, neurons plated on SynCAM1-coated substrates exhibit strongly patterned axons and selectively assemble functional presynapses. N-cadherin coating, however, is not able to elicit synapses, indicating the specificity of our system. This method opens the way to both fundamental and therapeutic studies of various synaptic systems.

Geometrically Mediated Topographic Steering of Neurite Behaviors and Network Formation

“…These studies indicate an active interaction of cells and processes with topographical features of varying scales. Specific arrangements of such spacing and gratings at the cell vicinity have been shown to elicit neurite guidance and initiate growth cone direction along predestined patterns . Moreover, chemical micropatterns are also known to induce cytoplasmic rearrangements resulting in the subsequent changes in neuritogenesis trends .…”

High-content imaging of presynaptic assembly

“…Our SV clusters detection script selectively and accurately identifies hemi-synapses that are induced by a synaptogenic cue expressed at the surface of a heterologous cell. Two recent papers describe the use of micropatterned substrates (Czondor et al, 2013) or specialized arrays of cells expressing synaptogenic adhesion molecules (Shi et al, 2011) to improve the sensitivity of this assay. Although our SV clusters detection script requires no specialized arrays or patterned substrates to reliably detect presynaptic induction, it can easily be combined with these innovative approaches.…”

Brain cells and neuronal networks: Encounters with controlled microenvironments

“…In this context, the approach developed by Czöndör et al [121] provided a novel strategy to achieve a precise distribution of synaptic contacts by substituting the post-synaptic structure by micro-dots coated with various synaptogenetic adhesion molecules (e.g. neurexin-1b, SynCAM1).…”