Membrane distribution of the glycine receptor alpha 3 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

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

15 Citations (Web of Science)

Abstract

In this study, the effect of glycine receptor (GlyR) ?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 ?3K and ?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 ?3L compared to ?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 ?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 ?3 membrane distribution, which has consequences for neuronal GlyR physiology and function.
Original languageEnglish
Pages (from-to)79-90
JournalHistochemistry and Cell Biology
Volume142
Issue number1
DOIs
Publication statusPublished - Jul 2014

Keywords

  • Super-resolution microscopy
  • Direct stochastic optical reconstruction microscopy
  • Pair correlation analysis
  • Glycine receptor
  • alpha 3 Subunit
  • RNA splicing

Cite this