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1 Division of Nephrology, University of Maryland School of Medicine, Baltimore, Maryland 21201-1595; and 2 Cardiovascular Research Institute, University of Leicester, Leicester LE1 7RH, United Kingdom
We investigated the dependence of ANG II
(10
8 M)-induced constriction of outer medullary
descending vasa recta (OMDVR) on membrane potential (
m) and chloride
ion. ANG II depolarized OMDVR, as measured by fully loading them with
the voltage-sensitive dye bis[1,3-dibutylbarbituric
acid-(5)] trimethineoxonol
[DiBAC4(3)] or selectively loading their pericytes. ANG
II was also observed to depolarize pericytes from a resting value of
55.6 ± 2.6 to 26.2 ± 5.4 mV when measured with
gramicidin D-perforated patches. When measured with
DiBAC4(3) in unstimulated vessels, neither changing
extracellular Cl concentration ([Cl]) nor
exposure to the chloride channel blocker indanyloxyacetic acid 94 (IAA-94; 30 µM) affected m. In contrast, IAA-94 repolarized OMDVR pretreated with ANG II. Neither IAA-94 (30 µM) nor niflumic acid (30 µM, 1 mM) affected the vasoactivity of unstimulated
OMDVR, whereas both dilated ANG II-preconstricted vessels. Reduction of
extracellular [Cl] from 150 to 30 meq/l enhanced ANG
II-induced constriction. Finally, we identified a Cl
channel in OMDVR pericytes that is activated by ANG II or by excision
into extracellular buffer. We conclude that constriction of OMDVR by
ANG II involves pericyte depolarization due, in part, to increased
activity of chloride channels.
medulla; kidney; microcirculation; membrane potential; patch clamp; niflumic acid; indanyloxyacetic acid 94
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