Analysis of NaCl transport in thin ascending limb of Henle's loop in CLC-K1 null mice.

To characterize the nature of NaCl transport in the thin ascending limb (tAL), we examined the transport properties of Na(+) and Cl(-) using in vitro microperfusion of the tAL in CLC-K1 null mice. In the presence of a transmural NaCl concentration gradient (100 mM higher in the lumen), the transepithelial diffusion voltage (V(d)) was 15.5 +/- 1.0 and -7.6 +/- 1.4 mV in CLC-K1(+/+) and CLC-K1(-/-) mice, respectively. Neither Cl(-) transport inhibitor 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB) nor acidification of the bathing fluid changed the V(d) values in CLC-K1(-/-) mice. The addition of 300 microg/ml protamine, a selective blocker of paracellular conductance, to the bath increased the V(d) values by 5.6 +/- 0.7 and 12.6 +/- 1.5 mV (P < 0.001) in CLC-K1(+/+) and CLC-K1(-/-) mice, respectively. Although efflux coefficients of (36)Cl were significantly decreased in CLC-K1(-/-) mice (188.3 +/- 25.6 in 4 tubules vs. 17.2 +/- 7.0 x 10(-5) cm/s in 6 tubules), those of (22)Na were not different between CLC-K1(+/+) and CLC-K1(-/-) mice. These results clearly indicate that the major component of Cl(-) transport sensitive to NPPB or pH is mediated by CLC-K1 in the tAL.

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