Regulation of Na+ channel density at the apical surface of rabbit urinary bladder epithelium

We have investigated the effects of various manipulations on Na+ transport across the rabbit urinary bladder epithelium. After bladders were mounted in Ussing chambers there was a spontaneous and significant (>4-fold) increase in amiloride-sensitive short-circuit current (equivalent to net Na+ transport) over a 6-h period. The increase in current was almost abolished by brefeldin A, an inhibitor of anterograde vesicular transport, and reduced after a 3-h delay by cycloheximide, an inhibitor of protein synthesis. The spontaneous increase in short-circuit current was potentiated by treatment of bladders with either forskolin, which causes an elevation in cAMP levels, or aldosterone. Acting together, these two agents produced a significant synergistic effect on short-circuit current. The short-circuit current recovered rapidly after reduction in intracellular Na+ levels, achieved either by lowering the extracellular Na+ concentration or blockade of epithelial Na+ channels with the sulphydryl modifying reagent p-chloromercuribenzenesulphonic acid (PCMBS). Recovery after PCMBS treatment was partially sensitive to brefeldin A. Short-circuit current saturated as the extracellular Na+ concentration was increased (EC50=51 trim). Saturation occurred over a range of Na+ concentrations in which single channel permeability is known to remain constant, indicating that it depends on a reduction in epithelial Na+ channel density at the apical plasma membrane. Exposure of bladders to a high Na+ concentration caused an increase in endocytotic activity, detected through an increase in the uptake of the fluid-phase marker fluorescein isothiocyanate (FITC)-dextran into vesicles located beneath the apical plasma membrane. We conclude that the urinary bladder epithelium is able to respond rapidly and efficiently to changes in its environment by regulating the density of epithelial Na+ channels in its apical surface. (C) 2002 Elsevier Science B.V All rights reserved.