Immortalized rabbit cortical collecting duct cells express AT(1) angiotensin II receptors
Immortalized rabbit cortical collecting duct cells express AT(1) angiotensin II receptors
复制标题
DOI：
10.1152/ajprenal.1996.271.6.f1147
复制
发表时间：
1996-12-01
影响因子：
4.2
通讯作者：
Hebert, RL
中科院分区：
文献类型：
--
作者：
Burns, KD;Regnier, L;Hebert, RL
将兔皮质集合管 (CCD) 细胞永生化，以研究 CCD 中的血管紧张素 II (ANG II) 信号传导。转染细胞保留了 CCD 特性；精氨酸加压素 (AVP)、前列腺素 E(2) 和异丙肾上腺素 (10(-7) M) 均显着刺激腺苷 3',5'-环单磷酸 (cAMP) 的产生；甲状旁腺激素、降钙素对cAMP无影响。 27% 的转染细胞结合 β 嵌入细胞标记花生凝集素，而针对主细胞和 α 嵌入细胞的抗体对 26% 的细胞进行免疫标记。所有细胞均用上皮细胞标记物细胞角蛋白的抗体染色。相比之下，平滑肌肌球蛋白、Tamm-Horsfall 蛋白或因子 VIII 的抗体未观察到免疫荧光。转染细胞表现出阿米洛利敏感的跨上皮短路电流。在转染细胞中，放射性配体结合测定检测到一类 ANG II 受体（亲和常数 = 0.78 nM），并且通过 Northern 分析证明了 AT(1) 受体 mRNA。 ANG II (10(-7) M)显着抑制AVP刺激的cAMP产生；较低浓度（10（-10）M）增加磷酸肌醇水解。总之，我们永生化了兔 CCD 细胞系，该细胞系保留了特征性的形态和激素特性。这些细胞表达 AT(1) 受体，与 cAMP 的抑制和磷酸肌醇周转的刺激相结合。我们假设这些信号通路可能介导 ANG II 对 CCD 运输和细胞生长的影响。
Rabbit cortical collecting duct (CCD) cells were immortalized to study angiotensin II (ANG II) signaling in the CCD. Transfected cells retained CCD properties; arginine vasopressin (AVP), prostaglandin E(2), and isoproterenol (10(-7) M) all significantly stimulated adenosine 3',5'-cyclic monophosphate (cAMP) production; and parathyroid hormone and calcitonin had no effect on cAMP. Twenty-seven percent of transfected cells bound the beta-intercalated cell marker peanut lectin agglutinin, whereas antibodies against principal cells and alpha-intercalated cells immunolabeled 26% of cells. All cells stained with antibodies to the epithelial cell marker cytokeratin. By contrast, no immunofluorescence was observed with antibodies to smooth muscle myosin, Tamm-Horsfall protein, or factor VIII. Transfected cells demonstrated amiloride-sensitive transepithelial short-circuit current. In transfected cells, radioligand binding assays detected a single class of ANG II receptors (affinity constant = 0.78 nM), and AT(1)-receptor mRNA was demonstrated by Northern analysis. ANG II (10(-7) M) significantly inhibited AVP-stimulated cAMP production; lower concentrations (10(-10) M) increased phosphoinositide hydrolysis. In summary, we immortalized a rabbit CCD cell line that retains characteristic morphological and hormonal properties. These cells express AT(1) receptors, coupled to inhibition of cAMP and to stimulation of phosphoinositide turnover. We postulate that these signaling pathways may mediate effects of ANG II on CCD transport and cell growth.