The present study used free-flow micropuncture and whole-kidney clearance studies to determine the renal response to normocalcemic vs. hypocalcemic acute volume expansion (AVE) in anesthetized Munich-Wistar rats. Animals received AVE with Ringer bicarbonate to 10% body weight; half of these animals were supplemented with calcium to maintain normocalcemia (VE + Ca2+) and half were allowed to become hypocalcemic (VE). Filtered load of chloride and total CO2 (TCO2) to the superficial proximal tubule and delivered load to the superficial loop segment were not different between groups. Superficial proximal tubule absolute Cl reabsorption was not different, but superficial loop segment absolute Cl reabsorption was less in the VE + Ca2+ animals (2,221 +/- 106 vs. 2,651 +/- 125 pmol/min, P less than 0.05) and whole-kidney fractional chloride excretion was greater (10.5 +/- 1.6 vs. 4.3 +/- 0.5%, P less than 0.05). When indomethacin (I) was administered to hypocalcemic (VE + I) and normocalcemic (VE + Ca2+ + I) AVE animals, both groups of animals had tubular and whole-kidney chloride reabsorption similar to VE animals. TCO2 reabsorption was not influenced by Ca2+ or I. The data indicate that normocalcemic vs. hypocalcemic AVE results in reduced superficial loop segment chloride reabsorption and greater whole-kidney fractional chloride excretion in the absence but not in the presence of prostaglandin inhibition. The data are compatible with an effect of hypocalcemia during AVE to limit superficial loop segment and whole-kidney chloride excretion by inhibiting renal prostaglandin synthesis.