In whole brain membranes of goldfish, 3H-chlorocyclopentyladenosine bound to adenosine A1 receptors. The A1 receptors were ubiquitously distributed in the brain with a maximum in the hypothalamus and a minimum in the spinal cord. In superfused goldfish cerebellar slices, cyclohexyladenosine inhibited the cyclic AMP accumulation stimulated by forskolin and the selective adenosine A1 receptor antagonist, 8-cyclopentyltheophylline, reversed this effect. 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, in a dose-dependent manner, inhibited the 30 mM K(+)-evoked release of glutamate whereas that of aspartate was unaffected. The CHA inhibition of glutamate-evoked release was reversed by 8-cyclopentyltheophylline. The adenosine A2a receptors were not detectable in whole brain membranes of goldfish either using the specific agonist 3H-CGS 21680 or 3H-5'-N-ethylcarboxamidoadenosine. The presence of A2b seems to be suggested by the NECA stimulation of cyclic AMP accumulation, which was reversed by 8-cyclopentyltheophylline. The results, taken together, indicate that adenosine has a neuromodulatory function in the nervous system of lower vertebrates which is comparable to that described in mammalian brain.