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Koji Eguchi Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Yuichiro Izumi Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Yukiko Yasuoka Department of Physiology, Kitasato University School of Medicine, Minami-ku, Sagamihara, Kanagawa, Japan

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Terumasa Nakagawa Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Makoto Ono Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Kosuke Maruyama Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Naomi Matsuo Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Akiko Hiramatsu Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Hideki Inoue Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Yushi Nakayama Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Hiroshi Nonoguchi Division of Internal Medicine, Kitasato University Medical Center, Kitamoto, Saitama, Japan

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Hyun-Wook Lee Division of Nephrology, Hypertension, and Transplantation, University of Florida College of Medicine, Gainesville, Florida, USA

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I David Weiner Division of Nephrology, Hypertension, and Transplantation, University of Florida College of Medicine, Gainesville, Florida, USA
Nephrology and Hypertension Section, North Florida/South Georgia Veterans Health System, Gainesville, Florida, USA

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Yutaka Kakizoe Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Takashige Kuwabara Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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Masashi Mukoyama Department of Nephrology, Kumamoto University Graduate School of Medical Sciences, Chuo-ku, Kumamoto, Japan

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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 Gӧ6983, 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.

Article Type:
Research Article
DOI:
https://doi.org/10.1530/JOE-20-0267
Volume/Issue:
Volume 249: Issue 2
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