In the present study we investigated the in vivo (acute and chronic) and in vitro effects of proline on NTPDase and 5'-nucleotidase activities in synaptosomes obtained from cerebral cortex of rats. For acute administration, 29-day-old rats received one subcutaneous injection of proline (18.2 micromol/g body weight) or an equivalent volume of 0.9% saline solution (control) and were killed 1 h later. For chronic treatment, buffered proline was injected subcutaneously into rats twice a day at 10 h intervals from the 6th to the 28th day of age. Rats were killed 12 h after the last injection. Results showed that acute and chronic proline administration provoked a reduction (25%) of ATP hydrolysis, but did not alter ADP and AMP hydrolysis. We also verified the in vitro effect of proline (3.0 microM-1.0 mM) on nucleotide hydrolysis in synaptosomes from cerebral cortex of rats. In contrast to the in vivo studies, it was not observed any statistically significant alteration on ATP, ADP and AMP hydrolysis. In conclusion, according to our results, it seems reasonable to postulate that proline administration alters the hydrolysis of ATP and probably affects the responses mediated by adenine nucleotides in the central nervous system of proline treated rats.