Confocal and TIRF microscopy based approaches to visualize arrestin trafficking in living cells
- Resource Type
- Authors
- Silvia Sposini; Frederic Jean-Alphonse
- Source
- Biomolecular Interactions Part A
Biomolecular Interactions Part A, 166, Elsevier, pp.179-203, 2021, Methods in Cell Biology, ⟨10.1016/bs.mcb.2021.06.009⟩
Biomolecular Interactions Part A ISBN: 9780128233511
- Subject
- Fluorescence-lifetime imaging microscopy
genetic structures
Endosome
Confocal
media_common.quotation_subject
Endosomes
Biology
law.invention
[SDV.BDLR.RS]Life Sciences [q-bio]/Reproductive Biology/Sexual reproduction
03 medical and health sciences
0302 clinical medicine
GPCR
Confocal microscopy
law
[SDV.BC.IC]Life Sciences [q-bio]/Cellular Biology/Cell Behavior [q-bio.CB]
Arrestin
Internalization
030304 developmental biology
media_common
G protein-coupled receptor
0303 health sciences
Trafficking
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology
[SDV.BBM.MN]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular Networks [q-bio.MN]
Subcellular localization
Cell biology
[SDV.SP.PHARMA]Life Sciences [q-bio]/Pharmaceutical sciences/Pharmacology
sense organs
030217 neurology & neurosurgery
TIRF microscopy
- Language
- English
Chapitre 8; International audience; Arrestins are key proteins that serve as versatile scaffolds to control and mediate G protein coupled receptors (GPCR) activity. Arrestin control of GPCR functions involves their recruitment from the cytosol to plasma membrane-localized GPCRs and to endosomal compartments, where they mediate internalization, sorting and signaling of GPCRs. Several methods can be used to monitor trafficking of arrestins; however, live fluorescence imaging remains the method of choice to both assess arrestin recruitment to ligand-activated receptors and to monitor its dynamic subcellular localization. Here, we present two approaches based on Total Internal Fluorescence (TIRF) microscopy and confocal microscopy to visualize arrestin trafficking in live cells in real time and to assess their co-localization with the GPCR of interest and their localization at specific subcellular locations.