Immunoneutralization of endogenous vasoactive intestinal peptide (VIP) by active immunization with chicken VIP (cVIP) reduced both basal circulating prolactin (PRL) and steady-state levels of pituitary PRL mRNA in the turkey. This immunoneutralization severely curtailed the plasma PRL response induced by infusion of cVIP into the median eminence, and totally blocked the plasma PRL release effected by electrical stimulation of the medial preoptic nucleus. This is the first demonstration that a stimulated PRL secretion can be blocked by neutralizing VIP availability. These findings imply that among the neurochemicals released by electrical stimulation, only VIP directly stimulates PRL secretion. In addition to serving as a PRL releasing factor, VIP also appears to be involved in the regulation of pituitary PRL mRNA expression.