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Am J Physiol Renal Physiol 282: F451-F457, 2002. First published August 30, 2001; doi:10.1152/ajprenal.0192.2001
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Vol. 282, Issue 3, F451-F457, March 2002

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

Wen Liu1, Tetsuji Morimoto1, Yoshiaki Kondo1, Kazuie Iinuma1, Shinichi Uchida2, Sei Sasaki2, Fumiaki Marumo2, and Masashi Imai3

1 Department of Pediatrics, Tohoku University School of Medicine, Sendai 980-8574; 2 Second Department of Internal Medicine, Tokyo Medical and Dental University, Tokyo 113-8519; and 3 Department of Pharmacology, Jichi Medical College, Kawachi 329-0498, Japan

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 (Vd) 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 Vd values in CLC-K1-/- mice. The addition of 300 µg/ml protamine, a selective blocker of paracellular conductance, to the bath increased the Vd 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 36Cl were significantly decreased in CLC-K1-/- mice (188.3 ± 25.6 in 4 tubules vs. 17.2 ± 7.0 × 10-5 cm/s in 6 tubules), those of 22Na 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.

gene targeting; renal medulla; urine-concentrating mechanism; chloride channel; paracellular shunt pathway; sodium chloride


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