Xenopus oocyte electrophysiology in GPCR drug discovery

Research output: Chapter in Book/Report/Conference proceedingBook chapterResearch

Deorphanization of the large group of G protein-coupled receptors (GPCRs) for which an endogenous activating ligand has not yet been identified (orphan GPCRs) has become increasingly difficult. A specialized technique that has been successfully applied to deorphanize some of these GPCRs involves two-electrode voltage-clamp recordings of currents through ion channels, which are activated by GPCRs heterologously expressed in Xenopus oocytes. The ion channels that couple to GPCR activation in Xenopus oocytes can be endogenous calcium-activated chloride channels (CaCCs) or heterologously expressed G protein-coupled inwardly rectifying potassium channels (GIRKs). We will describe a general approach for expression of GPCRs in Xenopus oocytes and characterization of these using electrophysiological recordings. We will focus on the detection of GPCR activation by recordings of currents through CaCCs that are activated by calcium release from the endoplasmic reticulum and thus the G(q) signaling pathway.
Original languageEnglish
Title of host publicationG protein-coupled receptors in drug discovery
EditorsWayne R Leifert
Volume552
Place of PublicationUnited States
PublisherHumana Press
Publication date2009
Pages343-357
ISBN (Print)978-1-60327-316-9
ISBN (Electronic)978-1-60327-317-6
DOIs
Publication statusPublished - 2009
SeriesMethods in Molecular Biology
Volume552
ISSN1064-3745

Bibliographical note

Keywords: Xenopus laevis, Oocyte, RNA, Transcription, Injection, GPRC6A, Intracellular calcium, Ligand.

Chapter 25

ID: 13063250