Fundamental role of ClC-3 in volume-sensitive Cl- channel function and cell volume regulation in AGS cells |
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Authors: | Jin Nan Ge Kim Jin Kyoung Yang Dong Ki Cho Soo Jin Kim Jung Mogg Koh Eun Ju Jung Hyun Chae So Insuk Kim Ki Whan |
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Affiliation: | Dept. of Physiology and Biophysics, Seoul National Univ. College of Medicine, 28 Yongon-Dong, Chongro-Gu, Seoul, Korea 110-799. |
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Abstract: | 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. |
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