Millimolar amiloride concentrations block K conductance in proximal tubular cells

Abstract

1 Amiloride, applied at millimolar concentrations, results in the blockade of K+ conductance in amphibian proximal convoluted cells (PCT), fused into giant cells. 2 Amiloride results directly in a blockade of K+ conductance that is not related to inhibition of the Na+-H+ antiport, which would lower intracellular pH, adversely affecting K+ conductance. On the contrary, high amiloride concentrations promote entry of this lipophilic base in the cell, leading to higher cell pH. 3 Under voltage clamp conditions, control vs. amiloride, current-voltage curves from PCT fused giant cells intersect at -86.2 ± 3.4 mV, a value close to the equilibrium potential for potassium. 4 Hexamethylene amiloride, 10-5 M, irreversibly depolarizes the membrane potential. 5 Barium decreased by 50% the initial slope of realkalinization, following removal of a solution containing NH4Cl, as did amiloride. In addition, these blockers reduced membrane conductance by 40%, suggesting that a fraction of the amiloride-suppressible NH4' efflux may be conductive. 6 Amiloride does not directly inhibit the Na+-K+, ATPase in our preparation, contrary to the prevalent -belief. 7 In vivo studies show that amiloride interferes with an apical K+ conductance but it does not alter basolateral K+ conductance.

Apical Location and Inhibition by Arginine Vasopressin of K+/H+ Antiport of the Medullary Thick Ascending Limb of Rat Kidney

“…First, the previously described MTAL NH 4 ϩ (Na ϩ ) channel (1), which is sensitive to M amiloride concentrations, transports K ϩ as well 4 and thus probably Rb ϩ . Second, residual quinidine-insensitive K ϩ channels, which may be blocked by high amiloride concentrations (1,30,31), could be responsible for the second component of conductive 86 Rb ϩ uptake which was blocked by 2 mM amiloride (Fig. 8).…”