Protocol to identify ligands of odorant receptors using two-electrode voltage clamp combined with the Xenopus oocytes heterologous expression system

Song Cao; Yang Liu; Guirong Wang

doi:10.1016/j.xpro.2022.101249

Protocol to identify ligands of odorant receptors using two-electrode voltage clamp combined with the Xenopus oocytes heterologous expression system

STAR Protoc. 2022 Mar 15;3(2):101249. doi: 10.1016/j.xpro.2022.101249. eCollection 2022 Jun 17.

Authors

Song Cao^{1

2}, Yang Liu¹, Guirong Wang^{1

2}

Affiliations

¹ State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
² Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, China.

Abstract

Two-electrode voltage clamp (TEVC) combined with the Xenopus laevis oocytes heterologous expression system is a powerful electrophysiological tool widely used to study the properties of many transmembrane proteins. Here, we describe a protocol using this combined approach to identify the ligands of odorant receptors that form ligand-gated ion channels. We detail the procedures for site-directed mutagenesis, oocyte microinjection, and TEVC recording. This protocol can also be used to identify the key residues and illustrate the structure-function relationships of these proteins. For complete details on the use and execution of this protocol, please refer to Cao et al. (2021).

Keywords: Gene Expression; Model Organisms; Molecular Biology; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Electrodes
Ligands
Oocytes / metabolism
Receptors, Odorant* / genetics
Xenopus laevis

Substances

Ligands
Receptors, Odorant