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Similar chloride channels in the connecting tubule and cortical collecting duct of the mouse kidney

Institut des Cordeliers, Laboratoire de Physiologie et Génomique des Cellules Rénales, UMR 7134, Université Pierre et Marie Curie and Centre National de la Recherche Scientifique, Paris, France

Submitted 4 July 2005 ; accepted in final form 9 January 2006

Using the patch-clamp technique, we investigated Cl^– channels on the basolateral membrane of the connecting tubule (CNT) and cortical collecting duct (CCD). We found a 10-pS channel in CNT cell-attached patches. Substitution of sodium gluconate for NaCl in the pipette shifted the reversal potential by +25 mV, whereas N-methyl-D-gluconate chloride had no effect, indicating anion selectivity. On inside-out patches, we determined a selectivity sequence of Cl^– > Br^– NO₃^– > F^–, which is compatible with that of ClC-K2, a Cl^– channel in the distal nephron. In addition, the number of open channels (NP_o) measured in cell-attached patches was significantly increased when Ca²⁺ concentration or pH in the pipette was increased, which is another characteristic of ClC-K. These findings suggest that the basis for this channel is ClC-K2. A similar Cl^– channel was found in CCD patches. Because CNT and CCD are heterogeneous tissues, we studied the cellular distribution of the Cl^– channel using recording conditions (KCl-rich solution in the pipette) that allowed us to detect simultaneously Cl^– channels and inwardly rectifying K⁺ channels. We detected Cl^– channels alone in 45% and 42% and K⁺ channels alone in 51% and 58% of CNT and CCD patches, respectively. Cl^– and K⁺ channels were recorded simultaneously from two patches (4% of patches) in the CNT and from none of the patches in the CCD. This indicates that Cl^– and K⁺ channels are located in different cell types, which we suggest may be the intercalated cells and principal cells, respectively.

ClC-K; renal tubule; intercalated cell

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