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While the overall, intracellular trafficking of the receptor was visualized continuously by using a GFP tag, selective imaging of cell surface receptors was achieved by pulse-labeling an acyl carrier protein tag. We found that OR17-40 efficiently translocated to the plasma membrane only at low expression, whereas at higher biosynthesis the receptor accumulated in intracellular compartments. Receptors in the plasma membrane showed high turnover resulting from constitutive internalization along the clathrin pathway, even in the absence of ligand. Single-molecule microscopy allowed monitoring of the early, dynamic processes in odorant receptor signaling. Although mobile receptors initially diffused either freely or within domains of various sizes, binding of an agonist or an antagonist increased partitioning of receptors into small domains of \u2248190 nm, which likely are precursors of clathrin-coated pits. The binding of a ligand, therefore, resulted in modulation of the continuous, constitutive internalization. After endocytosis, receptors were directed to early endosomes for recycling. This unique mechanism of continuous internalization and recycling of OR17-40 might be instrumental in allowing rapid recovery of odor perception.<\/jats:p>","DOI":"10.1073\/pnas.0603942103","type":"journal-article","created":{"date-parts":[[2006,9,16]],"date-time":"2006-09-16T00:41:26Z","timestamp":1158367286000},"page":"14325-14330","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":90,"title":["Visualizing odorant receptor trafficking in living cells down to the single-molecule level"],"prefix":"10.1073","volume":"103","author":[{"given":"V.","family":"Jacquier","sequence":"first","affiliation":[{"name":"Institute of Chemical Sciences and Engineering (ISIC), Ecole Polytechnique F\u00e9d\u00e9rale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland"}]},{"given":"M.","family":"Prummer","sequence":"additional","affiliation":[{"name":"Institute of Chemical Sciences and Engineering (ISIC), Ecole Polytechnique F\u00e9d\u00e9rale de 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