title: Hierarchical deconstruction of mouse olfactory sensory neurons: from whole mucosa to single-cell RNA-seq.
abstract: The mouse olfactory mucosa is a complex chemosensory tissue composed of multiple cell types, neuronal and non-neuronal. We have here applied RNA-seq hierarchically, in three steps of decreasing cellular heterogeneity: starting with crude tissue samples dissected from the nose, proceeding to flow-cytometrically sorted pools of mature olfactory sensory neurons (OSNs), and finally arriving at single mature OSNs. We show that 98.9% of intact olfactory receptor (OR) genes are expressed in mature OSNs. We uncover a hitherto unknown bipartition among mature OSNs. We find that 19 of 21 single mature OSNs each express a single intact OR gene abundantly, consistent with the one neuron-one receptor rule. For the 9 single OSNs where the two alleles of the abundantly expressed OR gene exhibit single-nucleotide polymorphisms, we demonstrate that monoallelic expression of the abundantly expressed OR gene is extremely tight. The remaining two single mature OSNs lack OR gene expression but express Trpc2 and Gucy1b2. We establish these two cells as a neuronal cell type that is fundamentally distinct from canonical, OR-expressing OSNs and that is defined by the differential, higher expression of 55 genes. We propose this tiered experimental approach as a paradigm to unravel gene expression in other cellularly heterogeneous systems.
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton-Cambridge, CB10 1SA, United Kingdom.|European Bioinformatics Institute (EMBL-EBI), European Molecular Biology Laboratory, Wellcome Genome Campus, Hinxton-Cambridge, CB10 1SD, United Kingdom.|Department of Experimental Genetics, Sidra Medical &Research Center, Qatar Foundation, PO Box 26999, Doha, Qatar.
authors.forename: Luis R
authors.initials: LR
authors.lastname: Saraiva
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton-Cambridge, CB10 1SA, United Kingdom.
authors.forename: Ximena
authors.initials: X
authors.lastname: Ibarra-Soria
authors.affiliation: Max Planck Research Unit for Neurogenetics, Max-von-Laue-Strasse 4, 60438 Frankfurt, Germany.
authors.forename: Mona
authors.initials: M
authors.lastname: Khan
authors.affiliation: Max Planck Research Unit for Neurogenetics, Max-von-Laue-Strasse 4, 60438 Frankfurt, Germany.
authors.forename: Masayo
authors.initials: M
authors.lastname: Omura
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton-Cambridge, CB10 1SA, United Kingdom.|European Bioinformatics Institute (EMBL-EBI), European Molecular Biology Laboratory, Wellcome Genome Campus, Hinxton-Cambridge, CB10 1SD, United Kingdom.
authors.forename: Antonio
authors.initials: A
authors.lastname: Scialdone
authors.affiliation: Max Planck Research Unit for Neurogenetics, Max-von-Laue-Strasse 4, 60438 Frankfurt, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Mombaerts
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton-Cambridge, CB10 1SA, United Kingdom.|European Bioinformatics Institute (EMBL-EBI), European Molecular Biology Laboratory, Wellcome Genome Campus, Hinxton-Cambridge, CB10 1SD, United Kingdom.
authors.forename: John C
authors.initials: JC
authors.lastname: Marioni
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton-Cambridge, CB10 1SA, United Kingdom.|Monell Chemical Senses Center, 3500 Market Street, Philadelphia, PA 19104, USA.
authors.forename: Darren W
authors.initials: DW
authors.lastname: Logan
mesh_terms: D000818:Animals; D016000:Cluster Analysis; D019295:Computational Biology / Q000379:methods; D005260:Female; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing; D008297:Male; D051379:Mice; D009831:Olfactory Mucosa / Q000166:cytology / Q000378:metabolism; D018034:Olfactory Receptor Neurons / Q000502:physiology*; D018035:Receptors, Odorant / Q000235:genetics; D059467:Transcriptome
chemical_list: D018035:Receptors, Odorant