title: Expression of glycine receptors in rat sensory neurons vs. HEK293 cells yields different functional properties.
abstract: Many structure-function studies of the glycine receptor (GlyR), and other ligand-gated ion channels, use somatic cell lines or Xenopus oocytes as expression systems. Using a polyethylenimine-based technique, we transfected GlyR cDNA into primary cultures of rat dorsal root ganglion (DRG) neurons. We then compared the functional properties of wildtype and a mutant GlyR expressed in DRG neurons with HEK 293 cells. The glycine sensitivity of the wildtype GlyR was nearly identical for the two cell types. The mutant GlyR has an arginine for glutamine substitution at position 271 (R271Q), which results in low glycine sensitivity relative to wildtype receptors expressed in HEK cells. This point mutation is associated with startle disease (hyperekplexia) in humans. Mutant GlyR expression in DRG neurons resulted in a significantly lower glycine sensitivity than was seen in HEK cells. This supports the idea that neuron-specific post-translational modifications may be important for determining receptor function.
authors.affiliation: Department of Anesthesia & Critical Care, University of Chicago, 5841 S, Maryland Avenue, MC4028, Chicago, IL 60637, USA.
authors.forename: A Y
authors.initials: AY
authors.lastname: Kung
authors.forename: C
authors.initials: C
authors.lastname: Rick
authors.forename: S
authors.initials: S
authors.lastname: O'Shea
authors.forename: N L
authors.initials: NL
authors.lastname: Harrison
authors.forename: D S
authors.initials: DS
authors.lastname: McGehee
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D005727:Ganglia, Spinal / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D009475:Neurons, Afferent / Q000378:metabolism / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018009:Receptors, Glycine / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology*; D014162:Transfection / Q000379:methods
chemical_list: D018076:DNA, Complementary; D018009:Receptors, Glycine