Xenopus Oocyte Electrophysiology in GPCR Drug Discovery
Xenopus Oocyte Electrophysiology in GPCR Drug Discovery
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DOI：
10.1007/978-1-60327-317-6_25
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发表时间：
2009-01-01
期刊：
影响因子：
--
通讯作者：
Brauner-Osborne, Hans
中科院分区：
文献类型：
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作者：
Hansen, Kasper B.;Brauner-Osborne, Hans
内源性激活配体尚未确定的大群G蛋白偶联受体（gpcr）（孤儿gpcr）的去孤儿化变得越来越困难。一种特殊的技术已经成功地应用于使这些gpcr的声音去孤儿化，该技术涉及通过离子通道的双电极电压钳记录电流，离子通道由非洲爪蟾卵母细胞中异种表达的gpcr激活。在爪蟾卵母细胞中，与GPCR激活偶联的离子通道可以是内源性钙激活的氯离子通道（CaCCs）或异源表达的G蛋白偶联的内向纠偏钾离子通道（GIRKs）。我们将描述一种在非洲爪蟾卵母细胞中表达gpcr的一般方法，并利用电生理记录对这些方法进行表征。我们将重点关注通过记录钙从内质网释放激活的CaCCs电流来检测GPCR激活，从而激活G(q)信号通路。
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 sonic 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.