Adenosine A2a receptors are not confined to dopamine-rich areas of the brain, since thermocycling analysis shows that adenosine A2a receptor mRNA is expressed also in the hippocampus (CA1, CA3 and dentate gyrus) and cerebral cortex. The expression of A2a mRNA in three main areas of the hippocampus was confirmed by in situ hybridization; A2a mRNA expression was mainly localized in the pyramidal and granular cells, the same hippocampal regions that showed adenosine A1 receptor mRNA expression. Receptor autoradiographic studies with [3H]CGS 21680 (30 nM), a selective adenosine A2a receptor agonist, showed specific binding sites in the hippocampus. The density of [3H]CGS 21680 binding was greatest in the stratum radiatum of the CA1 area, followed by the stratum oriens of the cornu Ammonis, stratum radiatum of the CA3 are and supra-granular layer of the dentate gyrus. This anatomical distribution of [3H]CGS 21680 binding was similar to the pattern of [3H]CHA binding in the hippocampus. Electrophysiological studies in the Schaffer fibers/CA1 pyramids showed that upon activation of the A2a receptors with CGS 21680 (10 nM) the ability of the adenosine A1 receptor agonist, CPA, to inhibit neuronal activity was significantly attenuated. These results show functionally important co-expression and co-localization of adenosine A2a and A1 receptors in the hippocampus. The results also suggest that adenosine A2a receptor-mediated neuromodulation is not confined to the basal ganglia, but is more widespread throughout the nervous system.