XB-ART-29566
J Biol Chem
1984 Sep 25;25918:11221-5.
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Aldosterone-stimulated sodium uptake by apical membrane vesicles from A6 cells.
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Sodium fluxes in plasma membrane vesicles prepared from the cultured toad kidney epithelial cell line A6 are studied. The vesicles are enriched 7-10 times in apical membrane markers. Sodium uptake is osmotically sensitive and inhibited by low concentrations of amiloride (K0.5 = 7 X 10(-8) M at 1 mM NaCl). Vesicles prepared from aldosterone-treated cells (4.5 h at 10(-7) M aldosterone) show a 2-fold enhancement of amiloride-sensitive sodium flux relative to appropriate controls. The above observations are in good agreement with studies of sodium transport across the apical membrane of intact A6 epithelia. Thus, the amiloride-sensitive sodium transporter in the apical membrane of these cells is preserved in the vesicle preparation, making it possible to study the effects of aldosterone in the absence of nonmembrane-related phenomena.
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