Odorant receptor proteins in the mouse main olfactory epithelium and olfactory bulb. (6th March 2017)

• Record Type: Journal Article
• Authors: Low, Victoria F.; Mombaerts, Peter
• Is Part Of: Neuroscience. Volume 344(2017)
• Page Start: 167
• Page End: 177
• Publication Date: 2017-03-06
• Subjects: aa amino acid -- AR antigen retrieval -- EDTA ethylenediaminetetraacetic acid -- GFP green fluorescent protein -- IRES internal ribosome entry site -- MOE main olfactory epithelium -- OMP olfactory marker protein -- OR odorant receptor -- OSN olfactory sensory neuron -- PBS phosphate-buffered saline -- PFA paraformaldehyde -- SDS sodium dodecyl sulfate; olfactory receptor -- olfactory sensory neuron -- glomerulus -- antibody -- immunofluorescence -- antigen retrieval; Neurochemistry -- Periodicals; Neurophysiology -- Periodicals; Neurology -- Periodicals; Neurochimie -- Périodiques; Neurophysiologie -- Périodiques; Neurochemistry; Neurophysiology; Electronic journals; Periodicals; Electronic journals; 612.8
• DOI: 10.1016/j.neuroscience.2016.12.044
• Languages: English
• ISSNs: 0306-4522

Abstract

Highlights: Odorant receptor proteins are expressed in mature olfactory sensory neurons of the mouse main olfactory epithelium. We screened 40 commercial antibodies against human and mouse ORs by immunofluorescence in the mouse main olfactory epithelium. Specificity and sensitivity of an antibody was evaluated for Olfr1507/MOR28 with a gene-targeted mouse strain. At least 20% of these antibodies are informative in immunohistochemical analyses of the mouse main olfactory epithelium. The commercial availability of these antibodies should facilitate the replicability of results in the literature. Abstract: In the mouse, odorant receptor proteins (ORs) are G-protein-coupled receptors expressed in mature olfactory sensory neurons (OSNs) of the main olfactory epithelium (MOE). ORs mediate odorant reception at the level of the OSN cilia. Most of the ∼1100 OR genes in the mouse genome are expressed, at the RNA level, in mature OSNs. The literature on antibodies against ORs is limited, and most reports are with antibodies that are not commercially available. Here we have screened 40 commercial antibodies against human and mouse ORs by immunofluorescence staining of coronal cryosections of the MOE of 21-day-old C57BL/6J mice. Various methods of antigen retrieval were tested. Of the 19 antibodies raised against human ORs, three yielded a consistent immunoreactive signal in the mouse MOE; of these three, two appeared to cross react against one or more, unknown, mouse ORs. Of the 21 antibodies raised against mouse ORs, six yielded a consistent immunoreactive signal in the mouse MOE; of these six, two also stained specific glomeruli in the olfactory bulb. Antibody specificity could be validated with gene-targeted mouse strains in the case of three ORs. The number of OSNs immunoreactive for the MOR28/Olfr1507 antibody is greater in C57BL/6J than in 129S6/SvEvTac wild-type mice. Taken together, our results are encouraging: 20–30% of these commercially available antibodies are informative in immunohistochemical analyses of the mouse MOE. The commercial availability of these antibodies should facilitate the study of OR proteins in the MOE and the olfactory bulb, and the replicability of results in the literature.