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  • 1
    Digitale Medien
    Digitale Medien
    Springer
    Pflügers Archiv 416 (1990), S. 533-539 
    ISSN: 1432-2013
    Schlagwort(e): MDCK-cells ; Aldosterone ; Na+/H+ exchange ; Cl−/HCO 3 − exchange ; Intercalated cells
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Experiments in dome epithelium of Madin-Darby canine kidney (MDCK) cells were performed to elucidate aldosterone action on acid-base transport. By means of pH-sensitive microelectrodes the pH of the dome fluid was measured while the apical plasma membrane was superfused. In the absence of HCO 3 − the dome fluid (facing the basolateral cell membrane) alkalinized in response to 10−7 mol/l aldosterone. Amiloride (10−3 mol/l) inhibited dome formation and pH recovery of the dome fluid from an extracellular acid load. In the presence of HCO 3 − dome fluid acidified in response to aldosterone. The stilbene derivative diisothiocyanate-stilbene-2,2′-disulphonic acid (DIDS) or removal of Cl− from the apical perfusate inhibited this dome acidification. In aldosterone-depleted MDCK monolayers HCO 3 − was actively accumulated in the dome fluid in contrast to aldosterone-supplemented cells. The results indicate that aldosterone stimulates both amiloride-sensitive Na+/H+ exchange and DIDS-sensitive Cl−/HCO 3 − exchange in the apical cell membrane of MDCK cells. In the absence of aldosterone the HCO 3 − extrusion process is localized in the basolateral membrane in series with apical Na+/H+ exchange, while in the presence of aldosterone Cl−/HCO 3 − is mainly localized in the apical membrane in parallel with Na+/H+ exchange. Cl− exits the cell through apical Cl− channels and is absorbed via the paracellular route.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    Digitale Medien
    Digitale Medien
    Springer
    Pflügers Archiv 416 (1990), S. 540-547 
    ISSN: 1432-2013
    Schlagwort(e): MDCK cells ; Omeprazole ; H+-K+ pump ; Carbonic anhydrase ; Intercalated cells
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: 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.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    Digitale Medien
    Digitale Medien
    Springer
    Pflügers Archiv 416 (1990), S. 526-532 
    ISSN: 1432-2013
    Schlagwort(e): Aldosterone ; MDCK cells ; Dome formation ; Differentiation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Medizin
    Notizen: Abstract Vectorial transport of salt and water in the Madin-Darby canine kidney (MDCK) cell line is indicated by the formation of domes when a monolayer is grown on an impermeable support. We investigated aldosterone-induced dome formation and evaluated the dome as an experimental model. Transepithelial dome resistance was about 80 Ωcm2 and constant when dome size exceeded 2 · 10−4 cm2. The relative ion conductances (expressed as transference numbers) across the dome epithelium were t Na∶t Cl∶t K= 0.64∶0.24∶0.06. They reflect the permeability properties of the paracellular shunt pathway tested at physiological concentrations of the individual ions. Aldosterone accelerated dome formation in serum-deprived MDCK monolayers. Prostaglandin E1 and transferrin were supportive but not essential for aldosterone-induced dome formation. After 72 h dome density was equal in monolayers cultured in serum-supplemented medium either in the presence or absence of mineralocorticoids. We conclude that aldosterone induces cell polarization in MDCK monolayers, leading to the formation of domes. The dome epithelium appears to be electrically isolated from the adjacent monolayer and can be studied by microelectrode techniques.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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