The mechanism of dopamine (DA) inhibition of pituitary PRL release is still unclear. To study it, we utilized enzymatically dispersed anterior pituitary cells obtained from adult female Sprague-Dawley rats. The cells were incubated in media with or without Na+ and in the presence or the absence of various drugs for 30 min for evaluating the secretion of PRL under baseline and experimental conditions. In some experiments, 45Ca2+ (1 microCi/ml) was added after 30 min of incubation and the latter prolonged for an additional minute to determine Ca2+ uptake. DA inhibited baseline PRL release and 45Ca2+ uptake in a dose-dependent manner only in the presence of Na+ and was totally inactive in its absence. The inhibitory effects of Nifedipine and Nicardipine, two Ca2+ channel antagonists, on PRL release were also found to be Na+ dependent. BAY K 8644, a Ca2+ channel agonist, stimulated PRL release and Ca2+ uptake in a dose-dependent manner, and these effects were enhanced by Na+-free media. DA antagonized the stimulatory actions of BAY K 8644 on PRL release in a similar dose-dependent manner both in the presence and the absence of Na+. However, on stimulated 45Ca2+ uptake DA was less effective in the absence of Na+. The stimulatory action of TRH on PRL release was enhanced by the absence of Na+. DA antagonized the effect of TRH in a dose-dependent manner both in the presence and in the absence of Na+ but appeared more effective in the absence of the ion. The PRL-releasing effects of phorbol ester and of the Ca2+ ionophore A23187 were antagonized by DA in a Na+- independent manner. These results suggest the existence of two mechanisms of DA inhibitory action: one exerted on baseline PRL release which is Na+ dependent, receptor linked, and probably implicates potential operated Ca2+ channels; the other is exerted on stimulated PRL release, is Na+ independent, and appears to be a postreceptorial intracellular event probably involving protein kinase C and/or cytosolic Ca2+ levels.