title: Multicolor time-resolved Förster resonance energy transfer microscopy reveals the impact of GPCR oligomerization on internalization processes.
abstract: Identifying the interacting partners and the dynamics of the molecular networks constitutes the key point in understanding cellular processes. Different methods often based on energy transfer strategies have been developed to examine the molecular dynamics of protein complexes. However, these methods suffer a couple of drawbacks: a single complex can be studied at a time, and its localization and tracking cannot generally be investigated. Here, we report a multicolor time-resolved Förster resonance energy transfer microscopy method that allows the identification of up to 3 different complexes simultaneously, their localization in cells, and their tracking after activation. Using this technique, we studied GPCR oligomerization and internalization in human embryonic kidney 293 cells. We definitively show that receptors can internalize as oligomers and that receptor coexpression deeply impacts oligomer internalization processes.
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Orestis
authors.initials: O
authors.lastname: Faklaris
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Martin
authors.initials: M
authors.lastname: Cottet
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Amandine
authors.initials: A
authors.lastname: Falco
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Brice
authors.initials: B
authors.lastname: Villier
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Laget
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Jurriaan M
authors.initials: JM
authors.lastname: Zwier
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Trinquet
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Mouillac
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France.
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Pin
authors.affiliation: *Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5203, Institut de Génomique Fonctionnelle, Montpellier, France; Institut National de la Santé et de la Recherche Médicale, Montpellier, France; Université Montpellier, Montpellier, France; Hamamastu Photonics France 19, Parc du Moulin de Massy, Massy, France; and Cisbio Bioassays, Codolet, France tdurroux@igf.cnrs.fr.
authors.forename: Thierry
authors.initials: T
authors.lastname: Durroux
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D055503:Protein Multimerization; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000378:metabolism*; D017483:Receptors, Vasopressin / Q000819:agonists / Q000737:chemistry / Q000378:metabolism; D015203:Reproducibility of Results; D059008:Time-Lapse Imaging / Q000295:instrumentation / Q000379:methods
chemical_list: D043562:Receptors, G-Protein-Coupled; D017483:Receptors, Vasopressin
keywords: lanthanide; multigate mode; oligomers; quantum dot; vasopressin V1a/V2 receptor