High-Throughput Odorant Receptor Deorphanization Via Phospho-S6 Ribosomal Protein Immunoprecipitation and mRNA Profiling.

Published

Journal Article

We describe an approach for the high-throughput surveying of odorant receptors (ORs) expressed in olfactory sensory neurons (OSNs) that have been activated by specific odorants. When OSNs are activated, there is a molecular signature in the form of a phosphorylated-S6 (pS6) ribosomal subunit. By the immunoprecipitation of the protein-RNA complex containing pS6, we identify the OR mRNA species expressed in these activated OSNs. The one neuron - one receptor rule (mature OSN expresses a single unique OR) allows for the identification of the collection of ORs that responded toward the tested odorant. Here we detail the procedure of (1) odor stimulation, (2) tissue harvesting, (3) immunoprecipitation, and (4) mRNA profiling for the high-throughput deorphanization of ORs in vivo.

Full Text

Duke Authors

Cited Authors

  • Hu, XS; Matsunami, H

Published Date

  • 2018

Published In

Volume / Issue

  • 1820 /

Start / End Page

  • 95 - 112

PubMed ID

  • 29884940

Pubmed Central ID

  • 29884940

Electronic International Standard Serial Number (EISSN)

  • 1940-6029

Digital Object Identifier (DOI)

  • 10.1007/978-1-4939-8609-5_8

Language

  • eng

Conference Location

  • United States