title: Membrane distribution of the glycine receptor α3 studied by optical super-resolution microscopy.
abstract: In this study, the effect of glycine receptor (GlyR) α3 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 α3K and α3L 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 α3L compared to α3K (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 μm(-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 α3 membrane distribution, which has consequences for neuronal GlyR physiology and function.
authors.affiliation: Biomedical Research Institute, Hasselt University and School of Life Sciences, Transnational University Limburg, Agoralaan Gebouw C, 3590, Diepenbeek, Belgium.
authors.forename: Kristof
authors.initials: K
authors.lastname: Notelaers
authors.forename: Susana
authors.initials: S
authors.lastname: Rocha
authors.forename: Rik
authors.initials: R
authors.lastname: Paesen
authors.forename: Nina
authors.initials: N
authors.lastname: Swinnen
authors.forename: Jeroen
authors.initials: J
authors.lastname: Vangindertael
authors.forename: Jochen C
authors.initials: JC
authors.lastname: Meier
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Rigo
authors.forename: Marcel
authors.initials: M
authors.lastname: Ameloot
authors.forename: Johan
authors.initials: J
authors.lastname: Hofkens
mesh_terms: D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D057809:HEK293 Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D012326:RNA Splicing / Q000235:genetics; D018009:Receptors, Glycine / Q000032:analysis* / Q000235:genetics / Q000378:metabolism
chemical_list: D018009:Receptors, Glycine; C113965:glycine receptor alpha3 subunit