Regulation of Rhcg, an ammonia transporter, by aldosterone in the kidney.
Regulation of Rhcg, an ammonia transporter, by aldosterone in the kidney.
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DOI：
10.1530/joe-20-0267
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发表时间：
2021-05
影响因子：
4
通讯作者：
Mukoyama, Masashi
中科院分区：
文献类型：
--
作者：
Eguchi, Koji;Izumi, Yuichiro;Yasuoka, Yukiko;Nakagawa, Terumasa;Ono, Makoto;Maruyama, Kosuke;Matsuo, Naomi;Hiramatsu, Akiko;Inoue, Hideki;Nakayama, Yushi;Nonoguchi, Hiroshi;Lee, Hyun-Wook;Weiner, I. David;Kakizoe, Yutaka;Kuwabara, Takashige;Mukoyama, Masashi
恒河猴C糖蛋白(Rhcg)是一种氨转运蛋白，是尿酸排泄的关键分子，主要表达于肾集合管间充质细胞(ICs)。在本研究中，我们研究了醛固酮在调节Rhcg表达中的作用。在用C57BL/6J小鼠进行的活体实验中，Western印迹分析表明，持续皮下注射醛固酮增加了肾膜部分Rhcg的表达。补钾可抑制醛固酮对Rh-cg的影响。接下来，在给予或不给予醛固酮的情况下，对小鼠进行肾上腺切除，然后随机给予含水的0.14M氯化铵。NH4Cl负荷增加了膜部分Rhcg的表达。肾上腺切除可降低NH4Cl诱导的Rhcg表达，但可通过给予醛固酮恢复。免疫组织化学研究显示，NH4Cl负荷可诱导Rhcg定位于外髓集合管内ICs的顶膜。肾上腺切除降低了NH4Cl诱导的Rhcg的膜定位，这一定位可通过给予醛固酮恢复。体外实验采用稳定表达FLAG标记的Rhcg(Rhcg-Flag)的永生化细胞系IN-IC细胞。Western印迹分析显示，醛固酮可增加膜组分中Rhcg-Flag的表达，而胞外钾水平的升高抑制了醛固酮的作用。螺内酯和PKC抑制剂Gö6983均抑制膜组分中Rhcg-Flag的表达。这些结果表明，醛固酮通过盐皮质激素受体和PKC途径调节RhcG的膜表达，该途径受细胞外钾水平的调节。
Rhesus C glycoprotein (Rhcg), an ammonia transporter, is a key molecule in urinary acid excretion and is expressed mainly in the intercalated cells (ICs) of the renal collecting duct. In the present study we investigated the role of aldosterone in the regulation of Rhcg expression. In in vivo experiments using C57BL/6J mice, Western blot analysis showed that continuous subcutaneous administration of aldosterone increased the expression of Rhcg in membrane fraction of the kidney. Supplementation of potassium inhibited the effect of aldosterone on the Rhcg. Next, mice were subjected to adrenalectomy with or without administration of aldosterone, and then ad libitum 0.14 M NH4Cl containing water was given. NH4Cl load increased the expression of Rhcg in membrane fraction. Adrenalectomy decreased NH4Cl-induced Rhcg expression, which was restored by administration of aldosterone. Immunohistochemical studies revealed that NH4Cl load induced the localization of Rhcg at the apical membrane of ICs in the outer medullary collecting duct. Adrenalectomy decreased NH4Cl-induced membrane localization of Rhcg, which was restored by administration of aldosterone. For in vitro experiments, IN-IC cells, an immortalized cell line stably expressing Flag-tagged Rhcg (Rhcg-Flag), were used. Western blot analysis showed that aldosterone increased the expression of Rhcg-Flag in membrane fraction, while the increase in extracellular potassium level inhibited the effect of aldosterone. Both spironolactone and Gö6983, a PKC inhibitor, inhibited the expression of Rhcg-Flag in the membrane fraction. These results suggest that aldosterone regulates the membrane expression of Rhcg through the mineralocorticoid receptor and PKC pathways, which is modulated by extracellular potassium level.
影响因子：
3.7
作者：
Izumi Y;Inoue H;Nakayama Y;Eguchi K;Yasuoka Y;Matsuo N;Nonoguchi H;Kakizoe Y;Kuwabara T;Mukoyama M
通讯作者：
Mukoyama M