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Am J Physiol Cell Physiol 280: C1511-C1520, 2001;
0363-6143/01 $5.00
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Vol. 280, Issue 6, C1511-C1520, June 2001

Expression and regulation of ClC-5 chloride channels: effects of antisense and oxidants

T. X. Weng1, L. Mo1, H. L. Hellmich2, A. S. L. Yu3, T. Wood4, and N. K. Wills1

Departments of 1 Physiology and Biophysics, 2 Internal Medicine, and 4 Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, Texas 77555; and 3 Renal Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02155

Genetic mutations of the Cl- channel ClC-5 cause Dent's disease in humans. We recently cloned an amphibian ortholog of Xenopus ClC-5 (xClC-5) from the A6 cell line. We now compare the properties and regulation of ClC-5 currents expressed in mammalian (COS-7) cells and Xenopus oocytes. Whole cell currents in COS-7 cells transfected with xClC-5 cDNA had strong outward rectification, Cl- > I- anion sensitivity, and were inhibited at low pH, similar to previous results in oocytes. In oocytes, antisense xClC-5 cRNA injection had no effect on endogenous membrane currents or the heterologous expression of human ClC-5. Activators of cAMP and protein kinase C inhibitors had no significant effects on ClC-5 currents expressed in either COS-7 cells or oocytes, whereas H-89, a cAMP-dependent protein kinase (PKA) inhibitor, and hydrogen peroxide decreased the currents. We conclude that the basic properties of ClC-5 currents were independent of the host cell type used for expression. In addition, ClC-5 channels may be modulated by PKA and reactive oxygen species.

Dent's disease; Xenopus oocytes; mammalian COS-7 cells; patch clamp; hydrogen peroxide


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