title: Time-resolved FRET between GPCR ligands reveals oligomers in native tissues.
abstract: G protein-coupled receptor (GPCR) oligomers have been proposed to play critical roles in cell signaling, but confirmation of their existence in a native context remains elusive, as no direct interactions between receptors have been reported. To demonstrate their presence in native tissues, we developed a time-resolved FRET strategy that is based on receptor labeling with selective fluorescent ligands. Specific FRET signals were observed with four different receptors expressed in cell lines, consistent with their dimeric or oligomeric nature in these transfected cells. More notably, the comparison between FRET signals measured with sets of fluorescent agonists and antagonists was consistent with an asymmetric relationship of the two protomers in an activated GPCR dimer. Finally, we applied the strategy to native tissues and succeeded in demonstrating the presence of oxytocin receptor dimers and/or oligomers in mammary gland.
authors.affiliation: Institut de Génomique Fonctionnelle, Centre National de la Recherche Scientifique, Montpellier, France.
authors.forename: Laura
authors.initials: L
authors.lastname: Albizu
authors.forename: Martin
authors.initials: M
authors.lastname: Cottet
authors.forename: Michaela
authors.initials: M
authors.lastname: Kralikova
authors.forename: Stoytcho
authors.initials: S
authors.lastname: Stoev
authors.forename: René
authors.initials: R
authors.lastname: Seyer
authors.forename: Isabelle
authors.initials: I
authors.lastname: Brabet
authors.forename: Thomas
authors.initials: T
authors.lastname: Roux
authors.forename: Hervé
authors.initials: H
authors.lastname: Bazin
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Bourrier
authors.forename: Laurent
authors.initials: L
authors.lastname: Lamarque
authors.forename: Christophe
authors.initials: C
authors.lastname: Breton
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Rives
authors.forename: Amy
authors.initials: A
authors.lastname: Newman
authors.forename: Jonathan
authors.initials: J
authors.lastname: Javitch
authors.forename: Eric
authors.initials: E
authors.lastname: Trinquet
authors.forename: Maurice
authors.initials: M
authors.lastname: Manning
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Pin
authors.forename: Bernard
authors.initials: B
authors.lastname: Mouillac
authors.forename: Thierry
authors.initials: T
authors.lastname: Durroux
mesh_terms: D000465:Algorithms; D000818:Animals; D065092:Antidiuretic Hormone Receptor Antagonists; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D019281:Dimerization; D065127:Dopamine D2 Receptor Antagonists; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes; D008024:Ligands; D008321:Mammary Glands, Animal / Q000378:metabolism; D008958:Models, Molecular; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism; D011869:Radioligand Assay; D051381:Rats; D017448:Receptors, Dopamine D2 / Q000819:agonists / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018045:Receptors, Oxytocin / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism; D017483:Receptors, Vasopressin / Q000819:agonists / Q000378:metabolism
chemical_list: D065092:Antidiuretic Hormone Receptor Antagonists; D065127:Dopamine D2 Receptor Antagonists; D005456:Fluorescent Dyes; D008024:Ligands; D009842:Oligopeptides; D017448:Receptors, Dopamine D2; D043562:Receptors, G-Protein-Coupled; D018045:Receptors, Oxytocin; D017483:Receptors, Vasopressin