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Membrane distribution of the glycine receptor α3 studied by optical super-resolution microscopy

Item Type:Article
Title:Membrane distribution of the glycine receptor α3 studied by optical super-resolution microscopy
Creators Name:Notelaers, K. and Rocha, S. and Paesen, R. and Swinnen, N. and Vangindertael, J. and Meier, J.C. and Rigo, J.M. and Ameloot, M. and Hofkens, J.
Abstract:In this study, the effect of glycine receptor (GlyR) {alpha}3 alternative RNA splicing on the distribution of receptors in the membrane of human embryonic kidney 293 cells is investigated using optical super-resolution microscopy. Direct stochastic optical reconstruction microscopy is used to image both {alpha}3K and {alpha}3L splice variants individually and together using single- and dual-color imaging. Pair correlation analysis is used to extract quantitative measures from the resulting images. Autocorrelation analysis of the individually expressed variants reveals clustering of both variants, yet with differing properties. The cluster size is increased for {alpha}3L compared to {alpha}3K (mean radius 92 ± 4 and 56 +/- 3 nm, respectively), yet an even bigger difference is found in the cluster density (9,870 ± 1,433 and 1,747 +/- 200 {mu}m(-2), respectively). Furthermore, cross-correlation analysis revealed that upon co-expression, clusters colocalize on the same spatial scales as for individually expressed receptors (mean co-cluster radius 94 +/- 6 nm). These results demonstrate that RNA splicing determines GlyR {alpha}3 membrane distribution, which has consequences for neuronal GlyR physiology and function.
Keywords:Super-Resolution Microscopy, Direct Stochastic Optical Reconstruction Microscopy, Pair Correlation Analysis, Glycine Receptor, {alpha}3 Subunit, RNA Splicing
Source:Histochemistry and Cell Biology
Page Range:79-90
Date:July 2014
Official Publication:https://doi.org/10.1007/s00418-014-1197-y
PubMed:View item in PubMed

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