title: Deciphering the Receptor Repertoire Encoding Specific Odorants by Time-Lapse Single-Cell Array Cytometry.
abstract: Mammals can recognize a vast number of odorants by using olfactory receptors (ORs) known as G protein-coupled receptors. The OR gene family is one of the most diverse gene families in mammalian genomes. Because of the vast combinations of ORs and odorants, few ORs have thus far been linked to specific odorants. Here, we established a functional screening method for OR genes by using a microchamber array containing >5,400 single olfactory epithelium-derived cells from mice applied to time-lapse single-cell array cytometry. This method facilitated the prompt isolation of single olfactory sensory neurons (OSNs) responding to the odorant of interest. Subsequent single-cell RT-PCR allowed us to isolate the genes encoding respective ORs. By using volatile molecules recognized as biomarkers for lung cancers, this method could deorphanize ORs and thereby reconstitute the OR-mediated signaling cascade in HEK293T cells. Thus, our system could be applied to identify any receptor by using specific ligands in the fields of physiopathology and pharmacology.
authors.affiliation: Advanced Research Division, Panasonic Corporation, 3-4 Hikaridai, Seika, Kyoto 619-0237, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Nobuo
authors.initials: N
authors.lastname: Yoshimoto
authors.affiliation: Advanced Research Division, Panasonic Corporation, 3-4 Hikaridai, Seika, Kyoto 619-0237, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Shimono
authors.affiliation: The Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Shun'ichi
authors.initials: S
authors.lastname: Kuroda
mesh_terms: D000818:Animals; D002469:Cell Separation / Q000379:methods*; D051379:Mice; D009812:Odorants; D009831:Olfactory Mucosa / Q000737:chemistry / Q000166:cytology; D018035:Receptors, Odorant / Q000737:chemistry* / Q000378:metabolism; D011984:Sensory Receptor Cells / Q000737:chemistry / Q000166:cytology; D046888:Tissue Array Analysis / Q000379:methods*
chemical_list: D018035:Receptors, Odorant
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