PUBLICATION

Functional integrity of green fluorescent protein conjugated glycine receptor channels

Authors
David-Watine, B., Shorte, S.L., Fucile, S., de Saint Jan, D., Korn, H., and Bregestovski, P.
ID
ZDB-PUB-990628-8
Date
1999
Source
Neuropharmacology   38(6): 785-792 (Journal)
Registered Authors
Bregestovski, Piotz, David-Watine, Brigitte, de Saint Jan, Didier, Korn, Henri
Keywords
glycine; GFP; imaging; receptor sorting; zebrafish; ion channel
MeSH Terms
  • Animals
  • Green Fluorescent Proteins
  • Indicators and Reagents
  • Luminescent Proteins/chemistry*
  • Microscopy, Fluorescence
  • Receptors, Glycine/chemistry*
  • Zebrafish
PubMed
10465682 Full text @ Neuropharmacology
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
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping