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Am J Physiol Renal Physiol 277: F251-F256, 1999;
0363-6127/99 $5.00
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Vol. 277, Issue 2, F251-F256, August 1999

Inhibition of initial transport rate of basolateral organic anion carrier in renal PT by BK and phenylephrine

Michael Gekle1, Sigrid Mildenberger1, Christoph Sauvant1, Dallas Bednarczyk2, Stephen H. Wright2, and William H. Dantzler2

1 Institute of Physiology, University of Würzburg, 97970 Würzburg, Germany; and 2 Department of Physiology, College of Medicine, University of Arizona, Tucson, Arizona 85721

The effect of ligands for phospholipase C-coupled receptors and of protein kinase C (PKC) stimulation with phorbol ester [phorbol 12-myristate 13-acetate (PMA)] or 1,2-dioctanoyl-sn-glycerol on the activity of the basolateral organic anion transporter (OAT) in S2 segments of single, nonperfused rabbit proximal tubules (PT) was measured with the use of fluorescein and epifluorescence microscopy. The initial uptake rate (25 s, OAT activity) was measured in real time by using conditions similar to those found in vivo. Stimulation of PKC with PMA or 1,2-dioctanoyl-sn-glycerol led to an inhibition of OAT activity, which could be prevented by 10-7 mol/l of the PKC-specific inhibitor bisindolylmaleimide. The alpha 1-receptor agonist phenylephrine as well as the peptide hormone bradykinin induced a reversible decrease of OAT activity, which was prevented by bisindolylmaleimide. The observed effect was not due to a decrease in the concentration of the counterion alpha -ketoglutarate or to impaired alpha -ketoglutarate recycling, because it was unchanged in the continuous presence of alpha -ketoglutarate or methyl succinate. We conclude that physiological stimuli can inhibit the activity of OAT in rabbit PT via PKC. The effect is not mediated by alterations in counterion availability but by a direct action on the OAT.

kidney; isolated proximal tubule; organic anion transport; protein kinase C; bradykinin


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