In goldfish brain, [3H]cyclohexyladenosine binding sites are ubiquitously distributed with a maximum in the hypothalamus and a minimum in the spinal cord. The binding parameters measured in cerebellar membranes (Kd = 0.88 +/- 0.08 nM; Bmax = 59.65 +/- 2.62 fmol/mg protein) are not significantly different from those of the whole brain. In perfused goldfish cerebellar slices, stimulation of cyclic AMP accumulation by 10(-5) M forskolin was markedly reduced (58.7%) by treatment with 10(-4) M cyclohexyladenosine, an adenosine A1 receptor agonist, and the reduction was reversed in the presence of 10(-4) M 8-cyclopentyltheophylline, a selective A1 receptor antagonist. In the same brain preparation, 30 mM K+ stimulated the release of glutamate, glutamine, glycine and GABA in a Ca(2+)-dependent manner, whereas the aspartate and taurine release was Ca(2+)-independent. Cyclohexyladenosine inhibited the 30 mM K(+)-evoked release of glutamate in a dose-related manner. This effect was reversed by 8-cyclopentyltheophylline. These results support the hypothesis that adenosine A1 receptors present in goldfish cerebellum are involved in the modulation of glutamate transmitter release.