Direct Visualization of Trans-Synaptic Neurexin-Neuroligin Interactions during Synapse Formation

作者：Tsetsenis Theodoros^*; Boucard Antony A; Arac Demet; Brunger Axel T; Suedhof Thomas C

来源：Journal of Neuroscience, 2014, 34(45): 15083-15096.

DOI：10.1523/JNEUROSCI.0348-14.2014

• microscopy
• neurexin
• neuroligin
• split-GFP
• synapses
• SynView

Neurexins and neuroligins are synaptic cell-adhesion molecules that are essential for normal synapse specification and function and are thought to bind to each other trans-synaptically, but such interactions have not been demonstrated directly. Here, we generated neurexin-1 beta and neuroligin-1 and neuroligin-2 fusion proteins containing complementary %26quot;split%26quot; GFP fragments positioned such that binding of neurexin-1 beta to neuroligin-1 or neuroligin-2 allowed GFP reconstitution without dramatically changing their binding affinities. GFP fluorescence was only reconstituted from split-GFP-modified neurexin-1 beta and neuroligin-1 if and after neurexin-1 beta bound to its neuroligin partner; reassociation of the split-GFP components with each other did not mediate binding. Using trans-cellular reconstitution of GFP fluorescence from split-GFP-modified neurexin-1 beta and neuroligins as an assay, we demonstrate that trans-synaptic neurexin/neuroligin binding indeed occurred when mouse hippocampal neurons formed synapses onto non-neuronal COS-7 cells expressing neuroligins or when mouse hippocampal neurons formed synapses with each other. This visualization of synapses by neurexin/neuroligin binding prompted us to refer to this approach as %26quot;SynView.%26quot; Our data demonstrate that neurexin-1 beta forms a trans-synaptic complex with neuroligin-1 and neuroligin-2 and that this interaction can be used to label synapses in a specific fashion in vivo.

• 出版日期2014-11-5