Abstract
gamma-aminobutyric acid type A (GABAA) receptors are the major sites of fast synaptic inhibition in the brain. They are constructed from four subunit classes with multiple members: alpha (1-6), beta (1-4), gamma (1-4), and delta (1). The contribution of subunit diversity in determining receptor subcellular targeting was examined in polarized Madin-Darby canine kidney (MDCK) cells. Significant detection of cell surface homomeric receptor expression by a combination of both immunological and electrophysiological methodologies was only found for the beta 3 subunit. Expression of alpha/beta binary combinations resulted in a nonpolarized distribution for alpha 1 beta 1 complexes, but specific basolateral targeting of both alpha 1 beta 2 and alpha 1 beta 3 complexes. The polarized distribution of these alpha/beta complexes was unaffected by the presence of the gamma 2S subunit. Interestingly, delivery of receptors containing the beta 3 subunit to the basolateral domain occurs via the apical surface. These results show that beta subunits can selectively target GABAA receptors to distinct cellular locations. Changes in the spatial and temporal expression of beta-subunit isoforms may therefore provide a mechanism for relocating GABAA receptor function between distinct neuronal domains. Given the critical role of these receptors in mediating synaptic inhibition, the contribution of different beta subunits in GABAA receptor function, may have implications in neuronal development and for receptor localization/clustering.
MeSH Terms
Animals
DNA, Complementary/administration & dosage
Dogs
Electrophysiology
Epitopes
Gene Expression
Kidney/cytology,metabolism
Microinjections
Microscopy, Phase-Contrast
Pentobarbital/pharmacology
Picrotoxin/pharmacology
Protein Conformation
Receptors, GABA-A/genetics,metabolism
Subcellular Fractions/metabolism
Chemicals
DNA, Complementary
Epitopes
Receptors, GABA-A
Picrotoxin
Pentobarbital
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Connolly C N
Medical Research Council Laboratory of Molecular Cell Biology, University College London, United Kingdom.
Wooltorton J R
Smart T G
Moss S J
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