In the male rat anterior pituitary, three morphological subtypes of cells secreting primarily prolactin (PRL) (lactotrophs) have been described. Type I contain predominantly large irregularly shaped granules, whereas type II and type III lactotrophs contain smaller spherical granules. We have previously shown that oestradiol and testosterone exert a rapid stimulatory effect selectively on type II lactotrophs but it is not known how the lactotroph subtypes respond to peptide secretagogues. We have therefore examined which cell subtype(s) release PRL in response to vasoactive intestinal peptide (VIP), thyrotrophin-releasing hormone (TRH) and prolactin-releasing peptide (PrRP-31). Pituitary segments were incubated in medium containing tannic acid (to capture exocytosis of secretory granules), either alone or with secretagogue peptide. VIP (1-10 nM), TRH (10 nM) and PrRP-31 (10 nM) all caused a significant increase (P < 0.05) in the amount of PRL granule exocytosis from type II and III lactotrophs, but had no effect on PRL exocytosis from type I. Dopamine (100 nM) inhibited basal exocytosis of immunoreactive (ir)-PRL from type I, II and III lactotrophs and PrRP-31-stimulated ir-PRL granule exocytosis from II and III lactotrophs. Treatment of lactating female rats with the dopamine D(2) receptor antagonist sulpiride (40 microg/kg) produced a significant increase (P < 0.05) in PRL granule exocytosis from type I and type III lactotrophs and a significant increase (P < 0.05) in the proportion of type I and II cells undergoing exocytosis of PRL. In conclusion, VIP, TRH and PrRP-31 selectively stimulate exocytosis from type II and III lactotrophs in the male rat, whereas all three lactotroph types are sensitive to dopamine inhibition of exocytosis in male and female rats.