When incubated in a tyrosine-free medium, the tissue dopamine (DA) level of rat striatal slices increased by about 921 +/- 15 pmol/mg protein during 90 min of preincubation. In contrast, the tissue-free tyrosine level declined only 130 pmol/mg protein in the same assay period. Depolarization of the slices with high K+ increased both DA and DOPAC outputs and depleted tissue DA level by about 75%. Although 60 min of resting after high K+ depolarization significantly restored the tissue DA levels, neither this restoration nor depolarization-induced DA release was altered by exogenous tyrosine. Similarly, failure of exogenous tyrosine was also observed during three successive depolarization periods of striatal slices. These results indicate that nigrostriatal dopaminergic neurons are able to synthesize and release the DA in the absence of exogenous tyrosine in the medium. Since the free tyrosine level in the slices does not seem to be a sufficient source, it is likely that tyrosine mobilized from its bound source(s) supports the DA synthesis under in vitro experimental conditions.