title: Arachidonic acid inhibits activity of cloned renal K+ channel, ROMK1.
abstract: Arachidonic acid (AA) has been shown to inhibit the activity of the low-conductance ATP-sensitive K+ channel in the apical membrane of the cortical collecting duct [W. Wang, A. Cassola, and G. Giebisch. Am. J. Physiol. 262 (Renal Fluid Electrolyte Physiol. 31): F554-F559, 1992]. ROMK1, a K+ channel derived from the rat renal outer medulla, shares many biophysical properties of the native low-conductance K+ channel, which is localized to the apical membranes of the cortical collecting duct and thick ascending limb. This study was designed to determine whether the ROMK channel maintains the property of AA sensitivity of the native low-conductance K+ channel. Experiments were conducted in Xenopus oocytes injected with cRNA encoding the ROMK1 channel by use of patch-clamp techniques. We have confirmed previous reports that the cloned ROMK1 has similar channel kinetics, high open probability, and inward slope conductance as the native low-conductance K+ channel, respectively. Addition of 5 microM AA to an inside-out patch resulted in reversible inhibition of channel activity at a concentration similar to the inhibitor constant for AA on the native K+ channel. The effect of AA on channel activity was preserved in the presence of 10 microM indomethacin, a cyclooxygenase inhibitor, 4 microM cinnamyl-3,4-dihydroxycyanocinnamate, a lipoxygenase inhibitor, and 4 microM 17-octadecynoic acid, an inhibitor of cytochrome P-450 monooxygenases, thus indicating that the effect of AA was not mediated by metabolites of AA. The effect did not appear to be the result of changes in membrane fluidity, since 5 microM eicosatetraynoic acid, an AA analogue that is a potent modulator of membrane fluidity, had no effect. Furthermore, the addition of AA to the outside of the patch also had no effect on channel activity. These results indicate that, like the native low-conductance channel, AA is able to directly inhibit ROMK1 channel activity.
authors.affiliation: Department of Pharmacology, New York Medical College, Valhalla 10595, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Macica
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: S C
authors.initials: SC
authors.lastname: Hebert
authors.forename: W H
authors.initials: WH
authors.lastname: Wang
mesh_terms: D000818:Animals; D016718:Arachidonic Acid / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D002109:Caffeic Acids / Q000494:pharmacology; D003001:Cloning, Molecular; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D065607:Cytochrome P-450 Enzyme Inhibitors; D005227:Fatty Acids / Q000494:pharmacology; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D005260:Female; D007213:Indomethacin / Q000494:pharmacology; D007679:Kidney Medulla / Q000378:metabolism*; D007700:Kinetics; D016859:Lipoxygenase Inhibitors / Q000494:pharmacology; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000235:genetics / Q000378:metabolism; D051381:Rats; D014982:Xenopus laevis
chemical_list: D002109:Caffeic Acids; D016861:Cyclooxygenase Inhibitors; D065607:Cytochrome P-450 Enzyme Inhibitors; D005227:Fatty Acids; D005231:Fatty Acids, Unsaturated; D016859:Lipoxygenase Inhibitors; D026902:Potassium Channel Blockers; D015221:Potassium Channels; C099255:cinnamyl-3,4-dihydroxycyanocinnamate; D016718:Arachidonic Acid; C047008:17-octadecynoic acid; D007213:Indomethacin