Magnesium and calcium transport was studied in the superficial proximal tubule, loop of Henle, and superficial distal tubule of the rat by in vivo microperfusion. Magnesium chloride was used to vary plasma and perfusate magnesium concentration. Perfusion with magnesium-free Ringer solution resulted in little magnesium entry into the tubule lumen in either normomagnesemic or hypermagnesemic rats. Magnesium concentration increased with water abstraction along the perfused proximal tubule whether markedly above or below plasma concentrations. Absolute proximal magnesium reabsorption increased with perfusate concentration; however, fractional magnesium and calcium reabsorption decreased in proportion to net sodium reabsorption with elevated extracellular magnesium. Magnesium absorption increased in the loop of Henle proportional to luminal magnesium concentration. Sodium and calcium reabsorption was not affected. Acute elevation of plasma magnesium, however, significantly depressed absolute magnesium reabsorption to a greater degree than calcium whereas sodium was unchanged. The effects of extracellular magnesium onloop reabsorption were reflected in the final urine. Reabsorption of sodium, calcium, and magnesium in the perfused distal tubule was concentration-dependent and not altered by luminal or extracellular magnesium concentration.