Guanine-based purines have been shown to modulate the effects of glutamate, which is essential for brain function and mediates excitotoxicity. In the search for a mechanism involving the interaction between purine nucleoside guanosine and glutamate, we found that guanosine dose-dependently, significantly (63%) and potently (EC50 =2.47 microM) enhanced glutamate uptake in cultured astrocytes. This effect was not inhibited by the blocker of nucleoside transporter dipyridamole nor by the adenosine antagonist theophylline, suggesting an extracellular site of action without the involvement of adenosine receptors. These results indicate a regulatory role of guanosine on extracellular levels of glutamate, possibly contributing for protecting neural cells against glutamate-induced excitotoxicity.