title: Dissociation and trafficking of rat GABAB receptor heterodimer upon chronic capsaicin stimulation.
abstract: Gamma-aminobutyric acid type B receptors (GABAB) are G-protein-coupled receptors that mediate GABAergic inhibition in the brain. Their functional expression is dependent upon the formation of heterodimers between GABAB1 and GABAB2 subunits, a process that occurs within the endoplasmic reticulum. However, the mechanisms that regulate GABAB receptor oligomerization at the plasma membrane remain largely unknown. We first characterized the functional cytoarchitecture of an organotypic co-culture model of rat dorsal root ganglia and spinal cord. Subsequently, we studied the interactions between GABAB subunits after chronic stimulation of sensory fibres with capsaicin. Surface labelling of recombinant proteins showed a decrease in subunit co-localization and GABAB2 labelling, after capsaicin treatment. In these conditions, fluorescence lifetime imaging measurements further demonstrated a loss of interactions between green fluorescent protein-GABAB1b and t-dimer discosoma sp red fluorescent protein-GABAB2 subunits. Finally, we established that the GABAB receptor undergoes clathrin-dependent internalization and rapid recycling to the plasma membrane following activation with baclofen, a GABAB agonist. However, in cultures chronically stimulated with capsaicin, the agonist-induced endocytosis was decreased, reflecting changes in the dimeric state of the receptor. Taken together, our results indicate that the chronic stimulation of sensory fibres can dissociate the GABAB heterodimer and alters its responsiveness to the endogenous ligand. Chronic stimulation thus modulates receptor oligomerization, providing additional levels of control of signalling.
authors.affiliation: INSERM U 862, Institut François Magendie, Université Bordeaux 2, Bordeaux, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Laffray
authors.forename: Kelly
authors.initials: K
authors.lastname: Tan
authors.forename: Josette
authors.initials: J
authors.lastname: Dulluc
authors.forename: Rabia
authors.initials: R
authors.lastname: Bouali-Benazzouz
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Calver
authors.forename: Frédéric
authors.initials: F
authors.lastname: Nagy
authors.forename: Marc
authors.initials: M
authors.lastname: Landry
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001640:Bicuculline / Q000494:pharmacology; D002211:Capsaicin / Q000494:pharmacology*; D018920:Coculture Techniques / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology; D018756:GABA Antagonists / Q000494:pharmacology; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology; D007476:Ionophores / Q000494:pharmacology; D008985:Monensin / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009475:Neurons, Afferent / Q000187:drug effects* / Q000502:physiology; D009924:Organ Culture Techniques; D021122:Protein Subunits / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D018080:Receptors, GABA-B / Q000378:metabolism*; D013116:Spinal Cord / Q000166:cytology / Q000502:physiology; D013997:Time Factors; D014162:Transfection / Q000379:methods
chemical_list: D018756:GABA Antagonists; D007476:Ionophores; D009419:Nerve Tissue Proteins; D021122:Protein Subunits; D018080:Receptors, GABA-B; D008985:Monensin; D002211:Capsaicin; D001640:Bicuculline