Deciphering the Receptor Repertoire Encoding Specific Odorants by Time-Lapse Single-Cell Array Cytometry

Scientific Reports
Masato SuzukiShun'ichi Kuroda

Abstract

Mammals can recognize a vast number of odorants by using olfactory receptors (ORs) known as G protein-coupled receptors. The OR gene family is one of the most diverse gene families in mammalian genomes. Because of the vast combinations of ORs and odorants, few ORs have thus far been linked to specific odorants. Here, we established a functional screening method for OR genes by using a microchamber array containing >5,400 single olfactory epithelium-derived cells from mice applied to time-lapse single-cell array cytometry. This method facilitated the prompt isolation of single olfactory sensory neurons (OSNs) responding to the odorant of interest. Subsequent single-cell RT-PCR allowed us to isolate the genes encoding respective ORs. By using volatile molecules recognized as biomarkers for lung cancers, this method could deorphanize ORs and thereby reconstitute the OR-mediated signaling cascade in HEK293T cells. Thus, our system could be applied to identify any receptor by using specific ligands in the fields of physiopathology and pharmacology.

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Citations

Apr 19, 2019·Lab on a Chip·Yusuke HirataShoji Takeuchi
Feb 19, 2019·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Hiroki OkayamaTomoyuki Yasukawa
May 7, 2021·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Misaki HataTomoyuki Yasukawa
Jun 17, 2021·Lab on a Chip·Yusuke HirataShoji Takeuchi

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Datasets Mentioned

BETA
AY317252
and
BC150839
AY317653
AY317250

Methods Mentioned

BETA
fluorescence-activated cell sorter
chip
PCR
nucleotide exchange
Single-Cell Array Cytometry

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