This autoradiographic study of sections of the rabbit stomach fundus labelled with [35S]dATP alpha S, a radioligand for P2Y purinoceptors, has demonstrated a discrete pattern of distribution of the binding sites, i.e., the specific binding was only over the mucosa, but not over the muscular layer. Radioligand binding assays carried out on gastric gland plasma membranes showed that the binding process was saturable and a high density of a homogeneous population of binding sites was observed. These binding sites presented high affinity with a value of Kd = 4.1 +/- 0.8 nM and the maximum density of the binding sites was 16.8 +/- 1.6 pmol/mg protein. The displacement by purinoceptor ligands showed the following order of potency: ATP = 2-methylthio ATP > > alpha, beta-methylene ATP > > adenosine. Neither UTP nor pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS) were able to displace the binding. The data support the presence of P2Y purinceptors in rabbit gastric glands.