title: Functional integrity of green fluorescent protein conjugated glycine receptor channels.
abstract: The alpha subunit (alphaZ1) of the zebrafish glycine receptor (GlyR) has been N-terminus fused with green fluorescent protein (GFP). We found that both pharmacological and electrophysiological properties of this chimeric alphaZ1-GFP are indistinguishable from those of the wild-type receptor when expressed in Xenopus oocytes and cell lines. The apparent affinities of this receptor for agonists (glycine, taurine and GABA), and the antagonist (strychnine) are unchanged, and single channel kinetics are not altered. In the same expression systems, alphaZ1-GFP was visualized using fluorescence microscopy. Fluorescence was distributed anisotropically across cellular membranes. In addition to the Golgi apparatus and endoplasmic reticulum, its presence was also detected on the plasmalemma, localized at discrete hot-spots which were identified as sites of high membrane turnover. Overall, the preservation in alphaZ1-GFPs of the wild type receptor functional properties makes it a promising new tool for further in situ investigations of GlyR expression, distribution and function.
authors.affiliation: INSERM U-261, Institut Pasteur, Paris, France. bdwati@pasteur.fr
authors.forename: B
authors.initials: B
authors.lastname: David-Watine
authors.forename: S L
authors.initials: SL
authors.lastname: Shorte
authors.forename: S
authors.initials: S
authors.lastname: Fucile
authors.forename: D
authors.initials: D
authors.lastname: de Saint Jan
authors.forename: H
authors.initials: H
authors.lastname: Korn
authors.forename: P
authors.initials: P
authors.lastname: Bregestovski
mesh_terms: D000818:Animals; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents; D008164:Luminescent Proteins / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D018009:Receptors, Glycine / Q000737:chemistry*; D015027:Zebrafish
chemical_list: D007202:Indicators and Reagents; D008164:Luminescent Proteins; D018009:Receptors, Glycine; D049452:Green Fluorescent Proteins