DETECTION OF ADENYLATE CYCLASE-COUPLED RECEPTORS IN XENOPUS OOCYTES BY COEXPRESSION WITH CYSTIC-FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR
DETECTION OF ADENYLATE CYCLASE-COUPLED RECEPTORS IN XENOPUS OOCYTES BY COEXPRESSION WITH CYSTIC-FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR
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DOI：
10.1006/abio.1995.1248
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发表时间：
1995-05-01
影响因子：
2.9
通讯作者：
ADAM, M
中科院分区：
文献类型：
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作者：
GRYGORCZYK, R;ABRAMOVITZ, M;ADAM, M
为了检测通过G蛋白偶联到非洲爪蟾卵母细胞中腺苷酸环化酶刺激的受体的异源表达，将感兴趣的受体与囊性纤维化跨膜传导调节因子（CFTR）-cAMP依赖性Cl-通道共表达。激动剂与表达的受体的结合刺激腺苷酸环化酶，导致细胞内cAMP升高，这反过来激活CFTR。然后使用标准双电极电压钳技术测量CFTR介导的Cl-电流响应。这种方法使我们能够检测人EP（2）和IP前列腺素受体在卵母细胞中的功能表达。这种方法应证明有价值的表达和鉴定的假定G蛋白偶联受体信号通过刺激腺苷酸环化酶，结构/功能的研究，并分析受体拮抗剂和激动剂。(C)出版社：Academic Press
To detect heterologous expression of receptors coupled via G proteins to the stimulation of adenylate cyclase in Xenopus laevis oocytes, the receptor of interest is coexpressed with the cystic fibrosis transmembrane conductance regulator (CFTR)-a cAMP-dependent Cl- channel. The binding of an agonist to the expressed receptor stimulates adenylate cyclase resulting in intracellular cAMP elevation, which in turn activates the CFTR. The CFTR-mediated Cl- current response is then measured using the standard two-electrode voltage-clamp technique. This method has allowed us to detect functional expression in oocytes of the human EP(2) and IP prostanoid receptors. This method should prove valuable for expression and identification of putative G protein-coupled receptors signaling through stimulation of adenylate cyclase, for structure/function studies, and for analysis of receptor antagonists and agonists. (C) 1995 Academic Press, Inc.