The hypocalciuric effect of thiazides: subcellular localization of the action

The acute administration of thiazides results in a decrease in the urinary Ca2+/Na+ ratio, whereas chronic administration of these diuretics decreases calciuria. In both situations, Ca2+ transport is enhanced in the early part of the distal tubule. The purpose of our study was to determine whether the hypocalciuric action of thiazides was due to a change in the active transport of Ca2+ through the basolateral membrane of the nephron or to an effect (direct or indirect) on the permeability of the distal tubule luminal membrane to calcium. In order to detect intrinsic differences between membranes of the proximal and distal tubules, the effect of the diuretic was examined in proximal and distal tubule preparations, and in basolateral and luminal membranes from the two segments separately. Preincubation of microdissected distal tubules in hypotonic solution containing 500 microM hydrochlorothiazide (HCTZ) did not influence the Ca2(+)-dependent ATP hydrolysis (Ca2+ = 1 microM) nor the Mg2(+)-dependent ATP hydrolysis (Mg2+ = 100 microM). Similarly 100 microM HCTZ did not change the Ca2+ ATPase activity in intact proximal and distal tubule suspensions, at Ca2+ concentrations ranging from 0.05 microM to 1 microM. ATP-dependent Ca2+ transport was present in basolateral membrane vesicles from proximal and distal tubule suspensions. Preincubation of the membranes with 100 microM HCTZ did not influence this transport. A Na+/Ca2+ exchanger, present in the basolateral membranes from the distal tubule, was also insensitive to HCTZ. In contrast, preincubation of luminal membranes from the distal tubules (but not proximal tubules) with 500 microM HCTZ significantly increased the Ca2+ uptake by these membranes.(ABSTRACT TRUNCATED AT 250 WORDS)