, Volume 416, Issue 5, pp 540-547

Madin-Darby canine kidney cells

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Abstract

Functionally and morphologically, Madin-Darby canine kidney (MDCK) cells resemble intercalated cells of urinary epithelia. Experiments were performed on domes of confluent MDCK monolayers to test for apical H+ secretion. Apical application of 10−3 mol/l amiloride or of Na+-free solution significantly reduced the limiting pH gradient across the dome epithelium (Δ pHd) consistent with inhibition of apical Na+/H+ exchange. Short-circuit current (SCC) measurements disclosed an acetazolamide-sensitive, (basolateral to apical) positive transepithelial current stimulated by 10−7 mol/l aldosterone and inhibited by acidification of apical medium to pH=4.5. Histochemical evaluation of carbonic anhydrase (CA) activity revealed cytoplasmic and apicalmembrane-bound CA particularly in dome-forming cells. Apical substitution of Na+ by K+ increased Δ pHd, whereas a reduction of K+ concentration to 0.5 mmol/l or addition of barium or omeprazole (10−5 mol/l) to the apical superfusate reduced Δ pHd by at least 75%. Aldosterone-stimulated SCC was completely abolished by the apical application of barium. We conclude that besides Na+/H+ exchange MDCK cells can express an apically located H+-K+ pump stimulated by aldosterone and inhibited directly by the anti-ulcer agent omeprazole or indirectly, either by blocking apical K+ recycling or by interfering with the CA-dependent intracellular formation of H+ ions.