title: In vivo identification of eugenol-responsive and muscone-responsive mouse odorant receptors.
abstract: Our understanding of mammalian olfactory coding has been impeded by the paucity of information about the odorant receptors (ORs) that respond to a given odorant ligand in awake, freely behaving animals. Identifying the ORs that respond in vivo to a given odorant ligand from among the ∼1100 ORs in mice is intrinsically challenging but critical for our understanding of olfactory coding at the periphery. Here, we report an in vivo assay that is based on a novel gene-targeted mouse strain, S100a5-tauGFP, in which a fluorescent reporter selectively marks olfactory sensory neurons that have been activated recently in vivo. Because each olfactory sensory neuron expresses a single OR gene, multiple ORs responding to a given odorant ligand can be identified simultaneously by capturing the population of activated olfactory sensory neurons and using expression profiling methods to screen the repertoire of mouse OR genes. We used this in vivo assay to re-identify known eugenol- and muscone-responsive mouse ORs. We identified additional ORs responsive to eugenol or muscone. Heterologous expression assays confirmed nine eugenol-responsive ORs (Olfr73, Olfr178, Olfr432, Olfr610, Olfr958, Olfr960, Olfr961, Olfr913, and Olfr1234) and four muscone-responsive ORs (Olfr74, Olfr235, Olfr816, and Olfr1440). We found that the human ortholog of Olfr235 and Olfr1440 responds to macrocyclic ketone and lactone musk odorants but not to polycyclic musk odorants or a macrocyclic diester musk odorant. This novel assay, called the Kentucky in vivo odorant ligand-receptor assay, should facilitate the in vivo identification of mouse ORs for a given odorant ligand of interest.
authors.affiliation: Department of Physiology, University of Kentucky, Lexington, Kentucky 40536, mcclint@uky.edu.
authors.forename: Timothy S
authors.identifier: http://orcid.org/0000-0002-4243-5323
authors.initials: TS
authors.lastname: McClintock
authors.affiliation: Department of Molecular Genetics and Microbiology and.
authors.forename: Kaylin
authors.initials: K
authors.lastname: Adipietro
authors.affiliation: Department of Physiology, University of Kentucky, Lexington, Kentucky 40536.
authors.forename: William B
authors.initials: WB
authors.lastname: Titlow
authors.affiliation: Department of Biostatistics, University of Iowa, Iowa City, Iowa 52242.
authors.forename: Patrick
authors.initials: P
authors.lastname: Breheny
authors.affiliation: Rockefeller University, New York, New York 10065.
authors.forename: Andreas
authors.initials: A
authors.lastname: Walz
authors.affiliation: Rockefeller University, New York, New York 10065, Max Planck Research Unit for Neurogenetics, D-60438 Frankfurt, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Mombaerts
authors.affiliation: Department of Molecular Genetics and Microbiology and Duke Institute for Brain Sciences, Department of Neurobiology, Duke University Medical Center, Durham, North Carolina 27710.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Matsunami
mesh_terms: D000818:Animals; D003516:Cycloparaffins / Q000494:pharmacology*; D005054:Eugenol / Q000494:pharmacology*; D006801:Humans; D008024:Ligands; D051379:Mice; D008810:Mice, Inbred C57BL; D009812:Odorants; D043562:Receptors, G-Protein-Coupled / Q000502:physiology; D018035:Receptors, Odorant / Q000187:drug effects*
chemical_list: D003516:Cycloparaffins; D008024:Ligands; D043562:Receptors, G-Protein-Coupled; D018035:Receptors, Odorant; D005054:Eugenol; C031021:muscone
keywords: G-protein coupled receptor; cell sorting; expression profiling; odor detection; olfaction; sensory coding