Tower-Gilchrist, CristyStyers, Melanie L.Yoder, Bradley K.Berbari, Nicolas F.Sztul, Elizabeth2023-09-192023-09-192014Tower-Gilchrist C, Styers ML, Yoder BK, Berbari NF, Sztul E. Monitoring endosomal trafficking of the g protein-coupled receptor somatostatin receptor 3. Methods Enzymol. 2014;534:261-280. doi:10.1016/B978-0-12-397926-1.00015-9https://hdl.handle.net/1805/35609Endocytic trafficking of G protein-coupled receptors (GPCRs) regulates the number of cell surface receptors available for activation by agonists and serves as one mechanism that controls the intensity and duration of signaling. Deregulation of GPCR-mediated signaling pathways results in a multitude of diseases, and thus extensive efforts have been directed toward understand the pathways and molecular events that regulate endocytic trafficking of these receptors. The general paradigms associated with internalization and recycling, as well as many of the key regulators involved in endosomal trafficking of GPCRs have been identified. This knowledge provides goalposts to facilitate the analysis of endosomal pathways traversed by previously uncharacterized GPCRs. Some of the most informative markers associated with GPCR transit are the Rab members of the Ras-related family of small GTPases. Individual Rabs show high selectivity for distinct endosomal compartments, and thus co-localization of a GPCR with a particular Rab informs on the internalization pathway traversed by the receptor. Progress in our knowledge of endosomal trafficking of GPCRs has been achieved through advances in our ability to tag GPCRs and Rabs with fluorescent proteins and perform live cell imaging of multiple fluorophores, allowing real-time observation of receptor trafficking between subcellular compartments in a cell culture model.en-USPublisher PolicySomatostatin receptor 3Endosomal traffickingLive cell imagingG protein-coupled receptor (GPCR)Rab GTPasesEpitope tagFluorescent proteinsArticle