Fundamental role of ClC-3 in volume-sensitive Cl- channel function and cell volume regulation in AGS cells.
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.
Jin
Nan Ge
NG
Dept. of Physiology and Biophysics, Seoul National Univ. College of Medicine, 28 Yongon-Dong, Chongro-Gu, Seoul, Korea 110-799.
Kim
Jin Kyoung
JK
Yang
Dong Ki
DK
Cho
Soo Jin
SJ
Kim
Jung Mogg
JM
Koh
Eun Ju
EJ
Jung
Hyun Chae
HC
So
Insuk
I
Kim
Ki Whan
KW
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics; D000906:Antibodies / Q000494:pharmacology; D055592:Biophysical Phenomena; D001703:Biophysics; D002460:Cell Line; D048429:Cell Size / Q000502:physiology; D018118:Chloride Channels / Q000037:antagonists & inhibitors / Q000235:genetics / Q000276:immunology / Q000502:physiology*; D002712:Chlorides / Q000037:antagonists & inhibitors; D004594:Electrophysiology; D004965:Estrogen Antagonists / Q000494:pharmacology; D005439:Flufenamic Acid / Q000494:pharmacology; D005753:Gastric Mucosa / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D007038:Hypotonic Solutions / Q000494:pharmacology; D007473:Ion Channels; D009579:Nitrobenzoates / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D013629:Tamoxifen / Q000494:pharmacology
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000906:Antibodies; C410035:CLNS1A protein, human; D018118:Chloride Channels; D002712:Chlorides; C115376:ClC-3 channel; D004965:Estrogen Antagonists; D007038:Hypotonic Solutions; D007473:Ion Channels; D009579:Nitrobenzoates; D012333:RNA, Messenger; D013629:Tamoxifen; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D005439:Flufenamic Acid; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid