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MUCOSAL BIOLOGY

Fundamental role of ClC-3 in volume-sensitive Cl^- channel function and cell volume regulation in AGS cells

¹Department of Physiology and Biophysics, Seoul National University College of Medicine, Seoul 110-799; ²Department of Anesthesiology, Sungkyunkwan University School of Medicine, Suwon 440-746; ³Department of Microbiology and Institute of Biomedical Science, Hanyang University College of Medicine, Seoul 33-791; ⁴Department of Internal Medicine, Liver Research Institute, Seoul National University College of Medicine, Seoul, Korea 110-744

Submitted 29 October 2002 ; accepted in final form 26 June 2003

Volume regulation is essential for cell function, but it is unknown which channels are involved in a regulatory volume decrease (RVD) in human gastric epithelial cells. Exposure to a hypotonic solution caused the increase in AGS cell volume, followed by the activation of a current. The reversal potential of the swelling-induced current suggested that Cl^- was the primary charge carrier. The selectivity sequence for different anions was I^- > Br^- > Cl^- > F^- > gluconate. This current was inhibited by flufenamate, DIDS, tamoxifen, and 5-nitro-2-(3-phenylpropylamino)benzoate. Intracellular dialysis of three different anti-ClC-3 antibodies abolished or attenuated the Cl^- current and disrupted RVD, whereas the current and RVD was unaltered by anti-ClC-2 antibody. Immunoblot studies demonstrated the presence of ClC-3 protein in Hela and AGS cells. RT-PCR analysis detected expression of ClC-3, MDR-1, and pICln mRNA in AGS cells. These results suggest a fundamental role of endogenous ClC-3 in the swelling-activated Cl^- channels function and cell volume regulation in human gastric epithelial cells.

swelling-activated chloride current; anti-ClC-3; gastric epithelial cell

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