Journal Article

Molecular profiling of activated olfactory neurons identifies odorant receptors for odors in vivo

Jiang, Yue, Hu, Xiaoyang Serene, Ni, Mengjue Jessica, Gong, Naihua Natalie, Pasi, Radhika, Matsunami, Hiroaki

2015

Overview

Using awake and freely behaving mice, this study employs a high-throughput in vivo RNA-seq approach to identify odorant receptor repertoires. The authors find sets of odorant receptors for two odorants encompassing 69 odorant receptor-odor pairs and develop models to predict receptor activation. The mammalian olfactory system uses a large family of odorant receptors (ORs) to detect and discriminate amongst a myriad of volatile odor molecules. Understanding odor coding requires comprehensive mapping between ORs and corresponding odors. We developed a means of high-throughput in vivo identification of OR repertoires responding to odorants using phosphorylated ribosome immunoprecipitation of mRNA from olfactory epithelium of odor-stimulated mice followed by RNA-Seq. This approach screened the endogenously expressed ORs against an odor in one set of experiments using awake and freely behaving mice. In combination with validations in a heterologous system, we identified sets of ORs for two odorants, acetophenone and 2,5-dihydro-2,4,5-trimethylthiazoline (TMT), encompassing 69 OR-odorant pairs. We also identified shared amino acid residues specific to the acetophenone or TMT receptors and developed models to predict receptor activation by acetophenone. Our results provide a method for understanding the combinatorial coding of odors in vivo .

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: 14/1 / 14/32 / 14/63 / 38/91 / 631/1647/2217/2018 / 631/378/2624/2625 / 631/378/340 / 64/60 / Acetophenone / Amino acids / Animal Genetics and Genomics / Animals / Behavioral Sciences / Biological Techniques / Biomedicine / Combinatorial analysis / Female / Gene expression / Gene Expression Profiling - methods / Genetic aspects / Genotype / High-Throughput Nucleotide Sequencing / Identification and classification / Immunohistochemistry / Immunoprecipitation / In Situ Hybridization / In vivo methods and tests / Ligands / Male / Messenger RNA / Methods / Mice / Mice, Inbred C57BL / mRNA / Neurobiology / Neurons - metabolism / Neurosciences / Odorant receptors / Odorants / Odors / Olfactory epithelium / Olfactory neurons / Olfactory Receptor Neurons - metabolism / Olfactory system / Phosphorylation / Polymerase Chain Reaction / Properties / Receptor mechanisms / Receptors / Receptors, Odorant - metabolism / Ribonucleic acid / RNA / RNA sequencing / Sensory receptors / Smell / Smell - physiology / Trimethylthiazoline