The effect of acute haloperidol administration on Fos protein expression was examined immunohistochemically in discrete regions of the rat brain. Male Wistar rats were injected subcutaneously (s.c.) with 0.1, 0.25, or 1.0 mg/kg of haloperidol. Two h after the injection, the rats were perfused, and the numbers of Fos immunoreactive neurons were counted in 24 brain regions. In contrast to the limited changes in Fos immunoreactivity at the low dose of haloperidol (0.1 mg/kg), the moderate dose (0.25 mg/kg) induced widespread increases in Fos-positive neurons in the rat brain. Large increases were produced in the caudate-putamen, nucleus accumbens, central amygdaloid nucleus, dorsomedial hypothalamic nucleus, hippocampus CA1 and substantia nigra pars compacta. Moderate increases were observed in the entorhinal cortex, lateral septum, lateral habenula, lateral amygdaloid nucleus, dentate gyrus, and mesencephalic central grey. Mild increases were induced in the anterior cingulate, temporal, occipital and perirhinal cortex, and central medial thalamic nucleus. The distribution of changes in Fos immunoreactivity at the high dose of haloperidol (1.0 mg/kg) were comparable to their distribution at the moderate dose. These findings indicate that the effect of acute haloperidol on Fos expression is widely distributed in the rat brain beyond the previously known dopamine-rich areas at the dose which produces plasma levels equivalent to those within the therapeutic range used clinically in humans. Further studies on the effects of chronic antipsychotic treatment are needed in order to identify the sites of the therapeutic action of antipsychotic drugs.