Apical membrane permeability and kinetic properties of the sodium pump in rabbit urinary bladder.
Previous studies have shown that aldosterone stimulates the rate of Na+ transport across the rabbit urinary bladder epithelium by increasing the apical membrane permeability to Na+. Paradoxically, ion-sensitive and conventional micro-electrode measurements demonstrated that intracellular Na+ activity aiNa+ was essentially unchanged by aldosterone, i.e. aiNa+ was constant regardless of the rate of Na+ transport. The present study was designed to resolve this apparent contradiction. The effects of elevated, endogenous aldosterone levels produced by low-Na+ diet (Lewis & Diamond, 1976) on urinary bladder Na+ transport were investigated in vitro using Ussing-type chambers and intracellular conventional and ion-sensitive microelectrodes. Apical membrane selectivity and kinetics of the Na+ pump were assessed as a function of hormone stimulation. The aldosterone-stimulated increase in Na+ transport was accounted for by increases in both the relative selective permeability of the apical membrane to Na+ and an increase in its absolute Na+ permeability. The kinetics of the Na+ pump were evaluated electrically by loading the cells with Na+ (monitored with Na+-sensitive micro-electrodes) or alternatively by manipulating serosal solution K+ concentration and measuring changes in the basolateral membrane electromotive forces and resistance. From these measurements the current generated by the pump was calculated as a function of intracellular Na+ or extracellular K+. The kinetics of the pump were not altered by aldosterone. A model of highly co-operative binding estimated Km for Na+ as 14.2 mM and 2.3 mM for K+. Hill coefficients for these ions were 2.8 and 1.8, respectively, consistent with a pump stoichiometry of 3 Na+ to 2 K+. The kinetic properties of the Na-K pump indicate that physiological levels of aiNa+ are poised at the foot of a step kinetic curve which energetically favours Na+ extrusion.
D000450:Aldosterone / Q000502:physiology; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D004039:Diet, Sodium-Restricted; D066298:In Vitro Techniques; D007473:Ion Channels; D007700:Kinetics; D008564:Membrane Potentials / Q000187:drug effects; D009761:Nystatin / Q000494:pharmacology; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D011817:Rabbits; D012964:Sodium / Q000378:metabolism*; D001743:Urinary Bladder / Q000378:metabolism* / Q000502:physiology
D007473:Ion Channels; D009761:Nystatin; D000450:Aldosterone; D010042:Ouabain; D012964:Sodium; D011188:Potassium