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Am J Physiol Renal Physiol 285: F326-F335, 2003. First published April 29, 2003; doi:10.1152/ajprenal.00034.2003
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Characterization of Ca2+-activated Cl– currents in mouse kidney inner medullary collecting duct cells

Zhiqiang Qu, Raymond W. Wei, and H. Criss Hartzell

Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322-3030

Submitted 23 January 2003 ; accepted in final form 24 April 2003

Ca2+-activated Cl– (ClCa) channels were characterized biophysically and pharmacologically in a mouse kidney inner medullary collecting duct cell line, IMCD-K2. Whole cell recording was performed with symmetrical N-methyl-D-glucamine chloride (NMDG)-Cl in the intracellular and extracellular solutions, and the intracellular Ca2+ concentration ([Ca2+]i) was adjusted with Ca2+-EGTA buffers. The amplitude of the current was dependent on [Ca2+]i. [Ca2+]i <800 nM strongly activated outwardly rectifying Cl– currents, whereas high Ca2+ (21 µM) elicited time-independent currents that did not rectify. The currents activated at low [Ca2+] exhibited time-dependent activation and deactivation. The affinity of the channel for Ca2+ was voltage dependent. The EC50 for Ca2+ was ~0.4 µM at +100 mV and ~1.0 µM at –100 mV. The Cl– channel blocker niflumic acid in the bath equally inhibited both inward and outward currents reversibly, with a Ki = 7.6 µM. DIDS, diphenylamine-2-carboxylic acid, and anthracene-9-carboxylic acid reversibly inhibited outward currents in a voltage-dependent manner. DTT slowly inhibited the currents, but tamoxifen did not. Comparing the biophysical and pharmacological properties, we conclude that IMCD-K2 cells express the same type of ClCa channels as those we have described in detail in Xenopus laevis oocytes (Qu Z and Hartzell HC. J Biol Chem 276: 18423–18429, 2001).

patch clamp; calcium; chloride channel; biophysics; pharmacology


Address for reprint requests and other correspondence: Z. Qu, Dept. of Cell Biology, 535 Whitehead Bldg., Emory Univ. School of Medicine, 615 Michael St., Atlanta, GA 30322-3030 (E-mail: zqu{at}cellbio.emory.edu).




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