Special Section on Super-resolution Microscopy of Neural Structure and Function

Nanoscale organization of synaptic adhesion proteins revealed by single-molecule localization microscopy

[+] Author Affiliations
Ingrid Chamma, Florian Levet, Jean-Baptiste Sibarita, Matthieu Sainlos, Olivier Thoumine

Centre National de la Recherche Scientifique, Interdisciplinary Institute for Neuroscience, UMR 5297, 147 rue Léo-Saignat, Bordeaux Cedex 33077, France

University of Bordeaux, Interdisciplinary Institute for Neuroscience, 147 rue Léo-Saignat, Bordeaux Cedex 33077, France

Neurophoton. 3(4), 041810 (Nov 03, 2016). doi:10.1117/1.NPh.3.4.041810
History: Received July 4, 2016; Accepted October 11, 2016
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Abstract.  The advent of superresolution imaging has created a strong need for both optimized labeling strategies and analysis methods to probe the nanoscale organization of complex biological structures. We present a thorough description of the distribution of synaptic adhesion proteins at the nanoscopic scale, namely presynaptic neurexin-1β (Nrx1β), and its two postsynaptic binding partners neuroligin-1 (Nlg1) and leucine-rich-repeat transmembrane protein 2 (LRRTM2). We monitored these proteins in the membrane of neurons by direct stochastic optical reconstruction microscopy, after live surface labeling with Alexa647-conjugated monomeric streptavidin. The small probe (3  nm) efficiently penetrates into crowded synaptic junctions and reduces the distance to target. We quantified the organization of the single-molecule localization data using a tesselation-based analysis technique. We show that Nlg1 exhibits a fairly disperse organization within dendritic spines, while LRRTM2 is organized in compact domains, and Nrx1β in presynaptic terminals displays a dual-organization pattern intermediate between that of Nlg1 and LRRTM2. These results suggest that part of Nrx1β interacts transsynaptically with Nlg1 and the other part with LRRTM2.

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© 2016 Society of Photo-Optical Instrumentation Engineers

Citation

Ingrid Chamma ; Florian Levet ; Jean-Baptiste Sibarita ; Matthieu Sainlos and Olivier Thoumine
"Nanoscale organization of synaptic adhesion proteins revealed by single-molecule localization microscopy", Neurophoton. 3(4), 041810 (Nov 03, 2016). ; http://dx.doi.org/10.1117/1.NPh.3.4.041810


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