title: High-throughput analysis of mammalian olfactory receptors: measurement of receptor activation via luciferase activity.
abstract: Odorants create unique and overlapping patterns of olfactory receptor activation, allowing a family of approximately 1,000 murine and 400 human receptors to recognize thousands of odorants. Odorant ligands have been published for fewer than 6% of human receptors(1-11). This lack of data is due in part to difficulties functionally expressing these receptors in heterologous systems. Here, we describe a method for expressing the majority of the olfactory receptor family in Hana3A cells, followed by high-throughput assessment of olfactory receptor activation using a luciferase reporter assay. This assay can be used to (1) screen panels of odorants against panels of olfactory receptors; (2) confirm odorant/receptor interaction via dose response curves; and (3) compare receptor activation levels among receptor variants. In our sample data, 328 olfactory receptors were screened against 26 odorants. Odorant/receptor pairs with varying response scores were selected and tested in dose response. These data indicate that a screen is an effective method to enrich for odorant/receptor pairs that will pass a dose response experiment, i.e. receptors that have a bona fide response to an odorant. Therefore, this high-throughput luciferase assay is an effective method to characterize olfactory receptors-an essential step toward a model of odor coding in the mammalian olfactory system.
authors: Monell Chemical Senses Center.
authors: Casey
authors: C
authors: Trimmer
authors: Monell Chemical Senses Center.
authors: Lindsey L
authors: LL
authors: Snyder
authors: Monell Chemical Senses Center; jmainland@monell.org.
authors: Joel D
authors: JD
authors: Mainland
mesh_terms: D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008024:Ligands; D008156:Luciferases / Q000032:analysis* / Q000378:metabolism; D009812:Odorants / Q000032:analysis; D018035:Receptors, Odorant / Q000032:analysis* / Q000378:metabolism
chemical_list: D008024:Ligands; D018035:Receptors, Odorant; D008156:Luciferases