Effects of Cl- transport inhibitors on Cl- permeability across hamster ascending thin limb

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Abstract
The highly conductive Cl- transport pathway exists in the ascending thin limb (ATL) of Henle's loop. To characterize the mechanism of the Cl- conductance across the ATL, we examined effects on Cl- permeability across hamster ATL of Cl- transport inhibitors, including 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), diphenylamine carboxylate (DPC), and anthracene-9-carboxylic acid (9-AC), by the in vitro microperfusion technique. NPPB added to the bath caused reversible suppression of the relative permeability of Cl- to Na+ (PCl/PNa), as estimated from the NaCl diffusion voltage in a dose-dependent manner in a range from 3 x 10(-6) to 10(-3) M. The concentration of NPPB that inhibited PCl/PNa by 50% (ID50) was approximately 3 X 10(-5) M. When 3 X 10(-5) M NPPB was added to the bath, the lumen-to-bath flux coefficient for 36Cl (Kl----b,Cl- 10(-7) cm2/s) was decreased from 130.7 +/- 7.3 to 52.2 +/- 11.6 (n = 7, P less than 0.01). Application of NPPB in the lumen also caused reversible suppression of PCl/PNa, but this effect was less potent compared with the application of the drug via the bath. Whereas 10(-3) M 9-AC or 10(-3) M DPC decreased PCl/PNa by 6.5 +/- 1.2 and 10.2 +/- 2.6%, respectively, 3 X 10(-4) M NPPB decreased PCl/PNa by 69.7 +/- 3.8%. In maleimide-treated tubules, addition of 10(-3) M N-ethylmaleimide (NEM) increased PCl/PNa from 1.0 +/- 0.1 to 1.8 +/- 0.1 (n = 7, P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

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