Renal and.
Department of Obstetrics and Gynecology, University of Iowa, Iowa City, Iowa.
J Am Soc Nephrol. 2018 Jun;29(6):1706-1719. doi: 10.1681/ASN.2017080826. Epub 2018 May 17.
is an E3 ubiquitin-protein ligase that associates with transport proteins, causing their ubiquitylation, and then internalization and degradation. Previous research has suggested a correlation between and BP. In this study, we explored the effect of intercalated cell (IC) gene ablation on IC transporter abundance and function and on BP. We generated IC knockout mice using Cre-lox technology and produced global pendrin/ null mice by breeding global null ( ) mice with global pendrin null ( ) mice. Mice ate a diet with 1%-4% NaCl; BP was measured by tail cuff and radiotelemetry. We measured transepithelial transport of Cl and total CO and transepithelial voltage in cortical collecting ducts perfused Transporter abundance was detected with immunoblots, immunohistochemistry, and immunogold cytochemistry. IC gene ablation markedly increased electroneutral Cl/HCO exchange in the cortical collecting duct, although benzamil-, thiazide-, and bafilomycin-sensitive ion flux changed very little. IC gene ablation did not increase the abundance of type B IC transporters, such as AE4 (), H-ATPase, barttin, or the Na-dependent Cl/HCO exchanger (). However, IC gene ablation increased CIC-5 total protein abundance, apical plasma membrane pendrin abundance, and the ratio of pendrin expression on the apical membrane to the cytoplasm. IC gene ablation increased BP by approximately 10 mm Hg. Moreover, pendrin gene ablation eliminated the increase in BP observed in global knockout mice. IC regulates Cl/HCO exchange in ICs., gene ablation increases BP in part through its action in these cells.
是一种 E3 泛素蛋白连接酶,可与转运蛋白结合,导致其泛素化,随后内吞和降解。先前的研究表明 与 BP 之间存在相关性。在这项研究中,我们探讨了间插细胞(IC)基因缺失对 IC 转运蛋白丰度和功能以及 BP 的影响。我们使用 Cre-lox 技术生成 IC 敲除小鼠,并通过将全局 敲除()小鼠与全局 pendrin 敲除()小鼠交配来产生全局 pendrin/敲除小鼠。小鼠食用含有 1%-4%NaCl 的饮食;BP 通过尾套和无线电遥测测量。我们测量了皮质集合管中 Cl 和总 CO 的跨上皮转运以及跨上皮电压。我们使用免疫印迹、免疫组织化学和免疫胶体金细胞化学检测转运体丰度。IC 基因缺失显著增加了皮质集合管中的电中性 Cl/HCO 交换,尽管苯甲脒、噻嗪类和巴弗洛霉素敏感的离子通量变化很小。IC 基因缺失并未增加 B 型 IC 转运体的丰度,如 AE4()、H-ATPase、barttin 或 Na 依赖性 Cl/HCO 交换体()。然而,IC 基因缺失增加了 CIC-5 总蛋白丰度、顶端质膜 pendrin 丰度以及顶端质膜上 pendrin 表达与细胞质的比值。IC 基因缺失使 BP 增加了约 10mmHg。此外,pendrin 基因缺失消除了在全局 敲除小鼠中观察到的 BP 增加。IC 调节 IC 中的 Cl/HCO 交换。IC 基因缺失增加了 BP,部分是通过其在这些细胞中的作用。