Interactions of monoamine, amino acid and cholinergic transmitter systems were studied in slices of rat cerebral cortex using a superfusion procedure and measuring release of endogenous dopamine (DA), norepinephrine (NE), serotonin (5-HT), GABA, glutamate (GLU) and aspartate (ASP). Depolarizing concentrations of K+ were used to induce a Ca2+-dependent, Mg2+-inhibited release of the monoamines and amino acids. Submaximal release of the monoamines and amino acids was observed at 35 mM K+, which permitted studies of possible excitatory or inhibitory actions of the added agents. The 35 mM K+-stimulated, Ca2+-dependent release of GABA was inhibited 40, 30 and 25% by 100 microM NE, DA and 5-HT, respectively. The release of GLU was potentiated by NE and reduced by DA. Both DA and 5-HT inhibited the release of ASP. The Ca2+-dependent, K+-stimulated release of endogenous NE, DA and 5-HT was not altered by 100 microM GABA, GLU or ASP. However, 100 microM GLU did enhance the stimulated release of GABA. The cholinergic agonist, carbachol, enhanced the stimulated release of NE, 5-HT and GLU 10, 60 and 40%, respectively. On the other hand, carbachol attenuated the release of DA and GABA approximately 20%. One interpretation of the data is that the amino acid transmitter pathways in slices of the cerebral cortex of the rat can be controlled by monoaminergic and cholinergic systems while the monoamine afferents appear to have a cholinergic regulation but not a major direct amino acid transmitter influence.