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Am J Physiol Renal Physiol 284: F977-F986, 2003. First published January 21, 2003; doi:10.1152/ajprenal.00255.2002
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Vol. 284, Issue 5, F977-F986, May 2003

Regulation by glucocorticoids and osmolality of expression of ROMK (Kir 1.1), the apical K channel of thick ascending limb

Morgan Gallazzini1, Amel Attmane-Elakeb1, David B. Mount2, Steven C. Hebert3, and Maurice Bichara1

1 Institut National de la Santé et de la Recherche Médicale U.426, Institut Fédératif Régional Claude Bernard, Faculté de Médecine Xavier Bichat, Université Paris 7, 75018 Paris, France; 2 Renal Division, West Roxbury Veterans Administration Medical Center and Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115; and 3 Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06520-8025

Mechanisms of regulation of ROMK channel mRNA and protein expression in medullary thick ascending limb (MTAL) were assessed in rat MTAL fragments incubated for 7 h. ROMK mRNA was quantified by quantitative RT-PCR and ROMK protein by immunoblotting analysis of crude membranes. Medium hyperosmolality (450 mosmol/kgH2O; NaCl plus urea added to isoosmotic medium) increased ROMK mRNA (P < 0.04) and protein (P < 0.006), and 10 nM dexamethasone also increased ROMK mRNA (P < 0.02). Hyperosmolality and dexamethasone had no additive effects on ROMK mRNA. NaCl alone, but not urea or mannitol, reproduced the hyperosmolality effect on ROMK mRNA. 1-Deamino-(8-D-arginine) vasopressin (1 nM) or 0.5 mM 8-bromo-cAMP had no effect per se on ROMK mRNA and protein. However, 8-bromo-cAMP abolished the stimulatory effect of dexamethasone on ROMK mRNA in the isoosmotic but not in the hyperosmotic medium (P < 0.004). In in vivo studies, the abundance of ROMK protein and mRNA increased in adrenalectomized (ADX) rats infused with dexamethasone compared with ADX rats (P < 0.02). These results establish glucocorticoids and medium NaCl concentration as direct regulators of MTAL ROMK mRNA and protein expression, which may be modulated by cAMP-dependent factors.

regulation of gene expression; ROMK channel; immunoblotting analysis; quantitative reverse transcriptase-polymerase chain reaction; isolated tubules


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