Expression of the chloride channel ClC-2 in the murine small intestine epithelium.
The chloride channel ClC-2 has been implicated in neonatal airway chloride secretion. To assess its role in secretion by the small intestine, we assessed its subcellular expression in ileal segments obtained from mice and studied the chloride transport properties of this tissue. Chloride secretion across the mucosa of murine ileal segments was assessed in Ussing chambers as negative short-circuit current (I(sc)). If ClC-2 contributed to chloride secretion, we predicted on the basis of previous studies that negative I(sc) would be stimulated by dilution of the mucosal bath and that this response would depend on chloride ion and would be blocked by the chloride channel blocker 5-nitro-2-(3-phenylpropylamino) benzoic acid but not by DIDS. In fact, mucosal hypotonicity did stimulate a chloride-dependent change in I(sc) that exhibited pharmacological properties consistent with those of ClC-2. This secretory response is unlikely to be mediated by the cystic fibrosis transmembrane conductance regulator (CFTR) channel because it was also observed in CFTR knockout animals. Assessment of the native expression pattern of ClC-2 protein in the murine intestinal epithelium by confocal and electron microscopy showed that ClC-2 exhibits a novel distribution, a distribution pattern somewhat unexpected for a channel involved in chloride secretion. Immunolabeled ClC-2 was detected predominantly at the tight junction complex between adjacent intestinal epithelial cells.
Gyömörey
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K
Programme in Cell Biology, Hospital for Sick Children, Toronto, Ontario, Canada M5G 1X8.
Yeger
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H
Ackerley
C
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Garami
E
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Bear
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CE
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D018118:Chloride Channels / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D005260:Female; D015870:Gene Expression / Q000502:physiology; D007038:Hypotonic Solutions / Q000494:pharmacology; D007082:Ileum / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D007413:Intestinal Mucosa / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018181:Mice, Inbred CFTR; D018345:Mice, Knockout; D008871:Microvilli / Q000737:chemistry / Q000378:metabolism; D009579:Nitrobenzoates / Q000494:pharmacology; D009997:Osmotic Pressure; D012333:RNA, Messenger / Q000032:analysis; D019108:Tight Junctions / Q000737:chemistry / Q000378:metabolism
D018118:Chloride Channels; D002712:Chlorides; D007038:Hypotonic Solutions; D009579:Nitrobenzoates; D012333:RNA, Messenger; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; C058176:5-nitro-2-(3-phenylpropylamino)benzoic acid; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid