title: Inhibition of ROMK channels by low extracellular K+ and oxidative stress.
abstract: We tested the hypothesis that low luminal K⁺ inhibits the activity of ROMK channels in the rat cortical collecting duct. Whole-cell voltage-clamp measurements of the component of outward K⁺ current inhibited by the bee toxin Tertiapin-Q (ISK) showed that reducing the bath concentration ([K⁺]o) to 1 mM resulted in a decline of current over 2 min compared with that observed at 10 mM [K⁺]o. However, maintaining tubules in 1 mM [K⁺]o without establishing whole-cell clamp conditions did not affect ISK. The [K⁺]o-dependent decline was not prevented by increasing cytoplasmic-side pH or by inhibition of phosphatase activity. It was, however, abolished by the inclusion of 0.5 mM DTT in the pipette solution to prevent oxidation of the intracellular environment. Conversely, treatment of intact tubules with the oxidant H₂O₂ (100 μM) decreased ISK in a [K⁺]o-dependent manner. Treatment of the tubules with the phospholipase C inhibitor U73122 prevented the effect of low [K⁺]o, suggesting the involvement of this enzyme in the process. We examined these effects further using Xenopus oocytes expressing ROMK2 channels. A 50-min exposure to the permeant oxidizing agent tert-butyl hydroperoxide (t-BHP; 500 μM) did not affect outward K⁺ currents with [K⁺]o = 10 mM but reduced currents by 50% with [K⁺]o = 1 mM and by 75% with [K⁺]o = 0.1 mM. Pretreatment of the oocytes with U73122 prevented the effects of t-BHP. Under conditions of low dietary K intake, K⁺ secretion by distal nephron segments may be suppressed by a combination of low luminal [K⁺]o and oxidative stress.
authors.affiliation: Department of Physiology and Biophysics, Weill-Cornell Medical College, New York, NY 10065, USA.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Frindt
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.forename: Henry
authors.initials: H
authors.lastname: Sackin
authors.forename: Lawrence G
authors.initials: LG
authors.lastname: Palmer
mesh_terms: D000818:Animals; D005260:Female; D066298:In Vitro Techniques; D007685:Kidney Tubules, Collecting / Q000378:metabolism; D008297:Male; D018384:Oxidative Stress / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000378:metabolism / Q000502:physiology*; D024661:Potassium Channels, Inwardly Rectifying / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014981:Xenopus
chemical_list: C510692:Kcnj1 protein, rat; D024661:Potassium Channels, Inwardly Rectifying; D011188:Potassium
keywords: K+ secretion; ROMK channels; U73122; oxidation; phospholipase C