The relationship between water and sodium movements through the mammalian proximal convoluted tubule was investigated by substituting lithium for sodium. Proximal convoluted rat Kidney tubules were perfused in vivo with a Ringer solution containing 107 meq/liter lithium and 42 meq/liter sodium. Several micropunctures were made along the same nephron, and [3H] inulin, [14C] glucose, 22Na, osmolality, Na, Mg and Cl were determined on each sample. Measurements of 22Na showed that sodium and lithium diffusion rates were practically identical throughout the entire epithelium. A one- for-one exchange of sodium for lithium induced a negative transepithelial net flux of Na from plasma to lumen. However, despite this negative flux, a positive net water movement was measured from lumen to plasma. This movement was proportional both to glucose reabsorption and to the rise in the chloride concentration, two mechanisms known to be dependent on the transcellular movement of sodium. It was therefore concluded that the net water flux was a function of the unidirectional transcellular net flux of Na. Rabbit proximal convoluted tubules were perfused in vitro with solution containing 75 meq/liter Li and 75 meq/liter Na on both the luminal and peritubular sides. Under these conditions, the water reabsorption rate dropped to half its control value. Water movement was therefore a function of the external sodium concentration, which in turn probably regulates the intracellular Na concentration.