The cellular localization of the uptake of [3H]adenosine and of binding of labelled adenosine analogues was studied in explant cultures of rat cerebellum and spinal cord by means of autoradiography. [3H]Adenosine was taken up by many neurons and astrocytes in both cerebellar and spinal cord cultures. The uptake of adenosine was inhibited in the absence of sodium or at 0 degrees C, suggesting an active transport mechanism. In both types of cultures, a great number of neurons showed binding sites for the A1-receptor agonist [3H]R-N6-phenylisopropyladenosine and for the mixed A1/A2-agonist [3H]N(ethyl)carboxamidoadenosine. Binding sites for both radioligands were also found on astrocytes, suggesting that these cells have receptors for the purinergic neurotransmitter adenosine. This suggestion is further supported by recent electrophysiological studies from our laboratory demonstrating that adenosine and its analogues produce hyperpolarizations of astrocytes which are blocked by the adenosine antagonist theophylline.