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

Am J Physiol Cell Physiol. 2001 Jun;280(6):C1511-20. doi: 10.1152/ajpcell.2001.280.6.C1511.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Anions / metabolism
  • Antisense Elements (Genetics)
  • Biological Transport / physiology
  • COS Cells
  • Chloride Channels / genetics*
  • Chloride Channels / metabolism*
  • Chlorides / metabolism
  • Cyclic AMP / metabolism
  • Cyclic AMP-Dependent Protein Kinases / antagonists & inhibitors
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Enzyme Inhibitors / pharmacology
  • Gene Expression / drug effects
  • Gene Expression / physiology
  • Humans
  • Hydrogen Peroxide / pharmacology*
  • Hydrogen-Ion Concentration
  • Isoquinolines / pharmacology
  • Kidney Diseases / metabolism
  • Membrane Potentials / drug effects
  • Membrane Potentials / physiology
  • Oocytes / physiology
  • Oxidants / pharmacology*
  • Patch-Clamp Techniques
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism
  • Sulfonamides*
  • Transfection
  • Xenopus

Substances

  • Anions
  • Antisense Elements (Genetics)
  • CLC-5 chloride channel
  • Chloride Channels
  • Chlorides
  • Enzyme Inhibitors
  • Isoquinolines
  • Oxidants
  • Sulfonamides
  • Hydrogen Peroxide
  • Cyclic AMP
  • Cyclic AMP-Dependent Protein Kinases
  • Protein Kinase C
  • N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide