Madin-Darby canine kidney cells

Pflügers Archiv - Tập 416 - Trang 540-547 - 1990
H. Oberleithner1, W. Steigner1, S. Silbernagl1, U. Vogel1, G. Gstraunthaler2, W. Pfaller2
1Department of Physiology, University of Würzburg, Würzburg, Germany
2Department of Physiology, University of Innsbruck, Innsbruck, Austria

Tóm tắt

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.

Tài liệu tham khảo

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