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Title: | Membrane distribution of the glycine receptor alpha 3 studied by optical super-resolution microscopy | Authors: | NOTELAERS, Kristof Rocha, Susana PAESEN, Rik SWINNEN, Nina Vangindertael, Jeroen Meier, Jochen C. RIGO, Jean-Michel AMELOOT, Marcel Hofkens, Johan |
Issue Date: | 2014 | Publisher: | SPRINGER | Source: | HISTOCHEMISTRY AND CELL BIOLOGY, 142 (1), p. 79-90 | 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. | Notes: | Rocha, S (reprint author), Katholieke Univ Leuven, Dept Chem, Lab Photochem & Spect, Celestijnenlaan 200F, B-3001 Louvain, Belgium. [Notelaers, Kristof; Paesen, Rik; Swinnen, Nina; Rigo, Jean-Michel; Ameloot, Marcel] Hasselt Univ, Biomed Res Inst, B-3590 Diepenbeek, Belgium. [Notelaers, Kristof; Paesen, Rik; Swinnen, Nina; Rigo, Jean-Michel; Ameloot, Marcel] Transnat Univ Limburg, Sch Life Sci, B-3590 Diepenbeek, Belgium. [Notelaers, Kristof; Rocha, Susana; Vangindertael, Jeroen; Hofkens, Johan] Katholieke Univ Leuven, Dept Chem, Lab Photochem & Spect, B-3001 Louvain, Belgium. [Meier, Jochen C.] Max Delbruck Ctr Mol Med, RNA Editing & Hyperexcitabil Disorders Grp, D-13092 Berlin, Germany. susana.rocha@chem.kuleuven.be; johan.hofkens@chem.kuleuven.be | Keywords: | Super-resolution microscopy; Direct stochastic optical reconstruction microscopy; Pair correlation analysis; Glycine receptor; alpha 3 Subunit; RNA splicing;super-resolution microscopy; direct stochastic optical reconstruction; microscopy; pair correlation analysis; glycine receptor; α3 subunit; RNA splicing | Document URI: | http://hdl.handle.net/1942/17056 | ISSN: | 0948-6143 | e-ISSN: | 1432-119X | DOI: | 10.1007/s00418-014-1197-y | ISI #: | 000338498700008 | Rights: | © Springer-Verlag Berlin Heidelberg 2014. | Category: | A1 | Type: | Journal Contribution | Validations: | ecoom 2015 |
Appears in Collections: | Research publications |
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