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Am J Physiol Heart Circ Physiol 281: H316-H324, 2001;
0363-6135/01 $5.00
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Vol. 281, Issue 1, H316-H324, July 2001

Nitric oxide generation by isolated descending vasa recta

Krisitie L. Rhinehart and Thomas L. Pallone

Division of Nephrology, University of Maryland School of Medicine, Baltimore, Maryland 21201-1595

Nitric oxide (NO) generation by the outer medullary descending vasa recta (OMDVR) was measured with the fluorescent dye 4,5-diaminofluoroscein (DAF-2) during 30-min incubations. Addition of 0.1 or 1.0 mM L-arginine to the incubation buffer increased the DAF-2 signal by 8.7 and 13.6% (P = 0.08 and P < 0.05), respectively. Compared with L-arginine alone (0.1 mM), bradykinin (BK; 1 × 10-7 M) enhanced the DAF-2 signal by 11.1% (P < 0.05). The NO synthase inhibitor Nomega -nitro-L-arginine methyl ester (0.1 mM) reversed the BK-stimulated NO generation as measured with either DAF-2 or by the oxidation of Fe2+ hemoglobin. Using 1 mM 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (tempol), a cell-permeant superoxide dismutase mimetic, we tested whether reduction of superoxide anion increases intracellular NO. Tempol increased DAF-2 fluorescence by 12 and 23.3%, respectively, over BK-stimulated or control vessels. Tempol also vasodilated ANG II (1 × 10-8 M)-preconstricted OMDVR (P < 0.05). We conclude that NO generation by isolated OMDVR can be increased by L-arginine, that the endothelium-dependent vasodilator BK enhances NO production, and that NO consumption by superoxide plays a role in the determination of cellular NO concentrations.

microperfusion; kidney; hemoglobin; fura 2; tempol; bradykinin; arginine


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