If we are to fully understand mechanisms of cartilage homeostasis, it is essential that we know the full catalogue of receptors present on the surface of a chondrocyte and the pathways regulated by ligands that bind to these receptors. In this study, we describe chondrocyte responses to adenosine 5'-triphosphate and related molecules. Adenosine 5'-triphosphate stimulated a statistically significant, dose-dependent, transient rise in the concentration of calcium ions in Fura 2-loaded, differentiated, primary chondrocytes. The increase occurred in the absence of extracellular calcium, indicating a mobilization from intracellular stores. The increase in concentration of cytoplasmic calcium ions induced by adenosine 5'-triphosphate was mimicked by uridine 5'-triphosphate but not by 2-methylthioadenosine 5'-triphosphate, cytidine 5'-triphosphate, or adenosine. Heterologous desensitization experiments demonstrated that chondrocytes showed no subsequent response to uridine 5'-triphosphate after initial stimulation with adenosine 5'-triphosphate nor did they respond to adenosine 5'-triphosphate in inverse conditions, thereby indicating competition for the same receptor site. Together, these results are consistent with the presence of a P2U receptor on the cell surface of chondrocytes. Purine-induced calcium mobilization in passaged chondrocytes showed the same pharmacological profile with respect to agonist sensitivity, but responses were of greater magnitude than responses in primary differentiated chondrocytes, suggesting upregulation of the receptor with time in culture. Adenosine 5'-triphosphate and uridine 5'-triphosphate (1-100 microM) did not alter cartilage matrix synthesis as measured by rate of incorporation of [35S]sulfate into glycosaminoglycan by cartilage explants or primary chondrocytes. Matrix degradation, measured by release of glycosaminoglycan from cartilage explants, was also unaltered by adenosine 5'-triphosphate or uridine 5'-triphosphate (1-100 microM). Production of prostaglandin E2 was upregulated by incubation with either adenosine 5'-triphosphate or uridine 5'-triphosphate. These data demonstrate the presence of a functional P2U-like purine receptor on the surface of primary articular chondrocytes and support the hypothesis that altered concentrations of extracellular purines may influence chondrocyte metabolism.