3 November 2016 Nanoscale organization of synaptic adhesion proteins revealed by single-molecule localization microscopy
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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
© 2016 Society of Photo-Optical Instrumentation Engineers (SPIE)
Ingrid Chamma, Ingrid Chamma, Florian Levet, Florian Levet, Jean-Baptiste Sibarita, Jean-Baptiste Sibarita, Matthieu Sainlos, Matthieu Sainlos, Olivier Thoumine, Olivier Thoumine, } "Nanoscale organization of synaptic adhesion proteins revealed by single-molecule localization microscopy," Neurophotonics 3(4), 041810 (3 November 2016). https://doi.org/10.1117/1.NPh.3.4.041810 . Submission:

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