Adenosine triphosphate (ATP) is a cotransmitter and an extracellular neuromodulator in nervous systems, and it influences neural circuits and synaptic strength. We have studied a stimulating effect of ATP (100 micro m) on the synaptic input of Purkinje neurons in acute cerebellar brain slices of juvenile rats (p14-19). Bath application of ATP increased the frequency of spontaneous postsynaptic currents (sPSCs) almost twofold, and increased their amplitude. These effects were fully suppressed by the P2 receptor antagonist pyridoxalphosphate-6-azophenyl-2'4'-disulphonic acid (PPADS; 10 microm), or after blocking action potentials with tetrodotoxin (TTX; 0.5 microm), but were not impaired by inhibiting ionotropic, non-NMDA glutamate receptors with 2,3-dioxo-6-nitro-1,2,3,4,-tetrahydrobenzo[f]quinoxaline-7-sulphonamide (NBQX; 5 microm). The frequency of sPSCs was reduced by 35% by PPADS and increased by 50% after inhibiting ectonucleotidase with ARL67156 (50 microm), suggesting intrinsic, 'tonic', stimulation of synaptic activity via P2 receptors. The pharmacological profile indicated that the ATP effect was mediated by both P2X and P2Y receptors, most probably of the P2X5- and P2Y(2,4)-like subtypes. The action potential frequency in the inhibitory basket cells was increased by 65%, and decreased in Purkinje neurons by 25%, in the presence of ATP. Our results suggest that ATP continuously modulates the cerebellar circuit by increasing the activity of inhibitory input to Purkinje neurons, and thus decreasing the main cerebellar output activity, which contributes to locomotor coordination.