Experiments were performed to document the presence of G protein-coupled P2Y nucleotide receptors in rat salivary glands and to examine changes in receptor expression during development and under conditions in which gland architecture is altered. The results indicate that, as opposed to mature rat submandibular gland (SMG), immature glands express functional P2Y1 receptors. P2Y1 receptor activity was highest at birth and declined over the next four weeks to undetectable levels. P2Y1 receptor mRNA levels remained constant over this time course, suggesting that receptor activity is regulated at some point other than transcription. Conversely, short-term culture of cells from the three major salivary glands resulted in upregulation of functional P2Y2 receptors. Responses to the P2Y2-selective agonist, UTP, were obtained after 3 h in culture and were maximal by 72 hours. This increase was paralleled by increased steady-state P2Y2 receptor mRNA levels. Upregulation of P2Y2 receptors also occurred in vivo following ligation of the main excretory duct of the SMG. These studies suggest that nucleotide receptors are dynamically regulated during development and as a result of perturbations to gland architecture.