Histochemistry and Cell Biology

, Volume 142, Issue 1, pp 79–90

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

  • Kristof Notelaers
  • Susana Rocha
  • Rik Paesen
  • Nina Swinnen
  • Jeroen Vangindertael
  • Jochen C. Meier
  • Jean-Michel Rigo
  • Marcel Ameloot
  • Johan Hofkens
Original Paper

DOI: 10.1007/s00418-014-1197-y

Cite this article as:
Notelaers, K., Rocha, S., Paesen, R. et al. Histochem Cell Biol (2014) 142: 79. doi:10.1007/s00418-014-1197-y

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.

Keywords

Super-resolution microscopyDirect stochastic optical reconstruction microscopyPair correlation analysisGlycine receptorα3 SubunitRNA splicing

Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Kristof Notelaers
    • 1
    • 2
  • Susana Rocha
    • 2
  • Rik Paesen
    • 1
  • Nina Swinnen
    • 1
  • Jeroen Vangindertael
    • 2
  • Jochen C. Meier
    • 3
  • Jean-Michel Rigo
    • 1
  • Marcel Ameloot
    • 1
  • Johan Hofkens
    • 2
  1. 1.Biomedical Research Institute, Hasselt University and School of Life SciencesTransnational University LimburgDiepenbeekBelgium
  2. 2.Laboratory for Photochemistry and Spectroscopy, Department of ChemistryKU LeuvenLouvainBelgium
  3. 3.RNA Editing and Hyperexcitability Disorders GroupMax Delbrück Center for Molecular MedicineBerlinGermany