It is becoming increasingly recognized that stimulation of apical P2 receptors can influence solute transport in the nephron, but, to date, no information is available on endogenous intraluminal nucleotide concentrations in vivo. This study measured intraluminal ATP concentrations in the renal tubules of anesthetized rats. Proximal tubular concentrations were found to be in the range of 100 to 300 nmol/L, with no significant variation along the S 2 segment, whereas concentrations in the early distal tubule were markedly lower. Using collections of varying duration, the half-life of ATP in collected proximal tubular fluid was found to be 3.4 min, indicating significant breakdown by soluble nucleotidases. For assessment of whether proximal tubular ATP was filtered or secreted, experiments were performed in Munich-Wistar rats. The ATP concentration in midproximal tubules (142 ؎ 23 nmol/L) was more than four-fold higher than in Bowman's space (32 ؎ 7 nmol/L; P < 0.001), whereas fractional water reabsorption between the two sites was modest. In experiments that were designed to determine the effects of (patho)physiologic disturbances on intraluminal ATP, rats were either volume expanded or subjected to hypotensive hemorrhage. Neither maneuver affected proximal tubular luminal ATP concentrations significantly; rapid degradation of secreted ATP by ecto-and soluble nucleotidases is a possible explanation. It is concluded that the proximal tubule secretes ATP into the lumen, where it may have an autocrine/paracrine regulatory role. T he epithelial cells of the mammalian nephron express a variety of P2Y and P2X receptor subtypes on their basolateral and apical membranes (1-3). When stimulated by application of exogenous nucleotides, these P2 receptors can influence a number of transport processes. Thus, activation of basolateral P2 receptors in the collecting duct inhibits vasopressin-stimulated water reabsorption (4 -6), and stimulation of apical P2 receptors can affect solute transport in both the proximal and the distal nephron: a recent in vivo micropuncture study in the rat showed that stimulation of apical P2Y 1 receptors inhibits proximal tubular bicarbonate reabsorption through suppression of NHE3 activity (7), and in vitro and in vivo evidence indicates that stimulation of apical P2 receptors in the distal nephron inhibits amiloride-sensitive sodium reabsorption (8,9) and reduces the activity of apical K ϩ secretory channels (10). These findings clearly support the proposal that intraluminal nucleotides, acting on apical P2 receptors, can function as autocrine/paracrine regulators of tubular transport. Before making such inferences concerning a functional role of intraluminal nucleotides, however, it is necessary to ascertain whether their normal endogenous concentrations are sufficient to activate P2 receptors and whether they can be influenced by physiologic/pathophysiologic maneuvers that are known to affect tubular transport. ATP secretion by renal epithelial cultures and by cell lines that were derived from ...