title: Characterization of sodium and chloride conductances in preneoplastic and neoplastic murine colonocytes.
abstract: Glucocorticoids, such as dexamethasone, induce amiloride-sensitive Na+ conductances in rat distal colon epithelium. The activity of these conductances diminishes from the surface to the base of the crypt whereas cAMP-stimulated Cl- secretion decreases from the crypt base to the surface. These gradients are likely to be perturbed during carcinogenesis. We therefore determined the magnitude of Na+ and Cl- conductances in colonocytes isolated from normal and carcinogen-treated rats. Colon carcinogenesis was induced by injection of dimethylhydrazine (DMH) (18 mg/kg) for 5 weeks. Before sacrifice animals were treated for 3 days with dexamethasone. Colonocyte populations from the surface to the crypt base (C1-C5) were harvested from the distal colon by a Ca2+-chelating procedure. The activity of Na+ conductances was determined by uptake of 22Na+ by surface and crypt colonocyte populations and by membrane vesicles in the presence and absence of 10 microM amiloride. In control rats Na+ conductance was highest in surface colonocytes and absent in the crypt base. As early as 2 weeks after initiation of DMH treatment amiloride-inhibited Na+ uptake was virtually absent in the upper crypt. Transcriptional assessment of the alpha-, beta- and gamma-subunits that constitute the epithelial Na+ channel revealed that DMH treatment reduces the expression of beta-subunit mRNA. We then examined 36Cl- efflux from isolated colonocytes of normal and carcinogen-treated rats in response to forskolin (0.01 mM). Forskolin induced a marked rise in cAMP in lower crypt cells concomitant with a significant stimulation of 36Cl- efflux. Intracellular cAMP increased in upper crypt cells in response to forskolin without an increase in 36Cl- efflux. By contrast, upper crypt colonocytes from DMH-treated rats showed forskolin-stimulated efflux beginning 4 weeks after initiation of treatment. We conclude that induction of Na+ conductances by glucocorticoids is inhibited during the early stages of chemical carcinogenesis due to lack of induction of the beta-subunit of the channel. By contrast, Cl- transport is stimulated both in surface and lower crypt cell compartments during different stages of chemical carcinogenesis.
authors.affiliation: Department of Gastroenterology, Rabin Medical Center (Beilinson Campus) and Sackler School of Medicine,
authors.forename: G M
authors.initials: GM
authors.lastname: Fraser
authors.forename: M
authors.initials: M
authors.lastname: Portnoy
authors.forename: M
authors.initials: M
authors.lastname: Bleich
authors.forename: D
authors.initials: D
authors.lastname: Ecke
authors.forename: Y
authors.initials: Y
authors.lastname: Niv
authors.forename: R
authors.initials: R
authors.lastname: Greger
authors.forename: B
authors.initials: B
authors.lastname: Schwartz
mesh_terms: D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D015152:Blotting, Northern; D002712:Chlorides / Q000502:physiology*; D005576:Colforsin / Q000494:pharmacology; D003106:Colon / Q000187:drug effects / Q000473:pathology / Q000503:physiopathology*; D003110:Colonic Neoplasms / Q000473:pathology / Q000503:physiopathology*; D000242:Cyclic AMP / Q000378:metabolism; D004553:Electric Conductivity; D007413:Intestinal Mucosa / Q000187:drug effects / Q000503:physiopathology; D011230:Precancerous Conditions / Q000473:pathology / Q000503:physiopathology*; D051381:Rats; D012016:Reference Values; D012964:Sodium / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*
chemical_list: D002712:Chlorides; D005576:Colforsin; D000584:Amiloride; D012964:Sodium; D000242:Cyclic AMP