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Ligands and signaling of the G-protein-coupled receptor GPR14, expressed in human kidney cells

机译:在人肾细胞中表达的配体和G蛋白偶联受体GPR14的信号传导

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Activation of the urotensin II (U-II) receptor, GPR14, leads to an increase in Ca2+, activation of phospholipase A(2) (PLA(2)) and an increase in arachidonic acid. The signaling pathway for guanylin peptides in the kidney involves an unknown G-protein coupled receptor which activates PLA(2) and increases arachidonic acid as well. To test if guanylin peptides could be, as U-II, agonists for the GPR14 receptor in the kidney, we used HEK293 and CHO cells transfected with hGPR14 (HEK293+hGPR14, CHO+hGPR14, respectively). Effects of guanylin peptides and U-II were studied by slow-whole-cell patch-clamp analysis and microfluorimetric measurements of intracellular Ca2+. Guanylin peptides and U-II depolarized HEK293+hGPR14 significantly more than wild type cells. These effects were inhibited in the presence of Ba2+ or PLA(2) inhibition (AACOCF(3)), suggesting that guanylin peptides and U-II increase arachidonic acid and inhibit ROMK channels in these cells. However, only U-II was capable to increase the cellular Ca2+, suggesting different mechanism of GPR14 activation by guanylin peptides and U-II. This signaling pathway of U-II involves PKC,because U-II effects in HEK293+hGPR14 cells were inhibited by calphostin C. Guanylin peptides activate PLA(2) and inhibit ROMK channels in HEK293 cells transfected with the human GPR14 receptor. Since GPR14 is present in mouse and human CCD it is a candidate for the guanylate cyclase independent receptor for guanylin peptides.
机译:urotensin II(U-II)受体GPR14的激活导致Ca2 +的增加,磷脂酶A(2)(PLA(2))的激活和花生四烯酸的增加。肾脏中鸟苷肽的信号传导途径涉及一个未知的G蛋白偶联受体,该受体激活PLA(2)并增加花生四烯酸。为了测试鸟苷肽是否可以作为肾脏GPR14受体的激动剂(如U-II),我们使用了用hGPR14(分别为HEK293 + hGPR14,CHO + hGPR14)转染的HEK293和CHO细胞。通过缓慢的全细胞膜片钳分析和细胞内Ca2 +的微荧光测量研究了鸟苷肽和U-II的作用。鸟苷肽和U-II对HEK293 + hGPR14的去极化作用明显超过野生型细胞。在Ba2 +或PLA(2)抑制(AACOCF(3))存在下,这些作用被抑制,表明鸟苷肽和U-II增加了花生四烯酸并抑制了这些细胞中的ROMK通道。但是,只有U-II能够增加细胞Ca2 +,表明鸟苷肽和U-II激活GPR14的机制不同。 U-II的此信号通路涉及PKC,因为钙磷蛋白C抑制了HEK293 + hGPR14细胞中的U-II效应。鸟苷肽激活了PLA(2)并抑制了转染人GPR14受体的HEK293细胞中的ROMK通道。由于GPR14存在于小鼠和人CCD中,因此它是鸟苷酸肽的鸟苷酸环化酶非依赖性受体的候选者。

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