Arachidonic acid potentiates the feedback response of large, ca2+ -activated k channels to angiotensin ii in human glomerular mesangial cells. Stockand, James D., Meredith Silverman, David Hall, Thomas Derr, Brian Kubacak, and Steven C. Sansom. Departments of Medicine and Integrative Biology, Pharmacology and Physiology, University of Texas Medical School at Houston and Department of Physiology, and Biophysics, University of Nebraska Medical Center
APStracts 5:0008F, 1998.
The influence of arachidonic acid (AA) on the feedback regulation of mesangial contraction by large Ca2+-activated K+ channels (BKCa) was determined through single channel analysis using the patch clamp method. The mesangial BKCa is a low gain negative feedback inhibitor of contraction that is activated in response to agonist-induced Ca2+ transients and membrane depolarization. Arachidonic acid activated BKCa in cell attached patches in a dose-dependent manner with a maximal effect at 400 nM and a half maximal response at 49 nM. In inside out patches, AA directly activated BKCa with a maximal effect at 400 nM. BKCa was activated significantly in response to addition of 100 nM angiotensin II (ANGII) in the presence but not the absence of AA. Since it was shown previously that fatty acids stimulated both soluble and membrane-bound guanylyl cyclase, we determined if AA activated BKCa by interfering with guanosine 3',5'-cyclic monophosphate (cGMP)-mediated signal transduction pathways. It was previously shown that 10 [mu]M cGMP, via cGMP-dependent protein kinase, activated BKCa in a biphasic manner with an early increase in Po and a subsequent inactivation mediated by phosphoprotein phosphatase 2A (PP2A). We found that 10 [mu]M dibutyryl cGMP enhanced BKCa activity in an additive manner with saturating concentrations (400 nM) of AA. Moreover, the inactivation phase mediated by PP2A was not abolished. Thus, AA does not affect the phosphorylation/dephosphorylation regulatory cycle for BKCa. It is concluded that AA potentiates the ANGII feedback response of BKCa by a mechanism that is independent of the phosphorylation cycle.

Received 2 May 1997; accepted in final form 9 January 1998.
APS Manuscript Number F150-7.
Article publication pending Am. J. Physiol. (Renal Physiology).
ISSN 1080-4757 Copyright 1998 The American Physiological Society.
Published in APStracts on 28 January 1998