Pituitary adenylate cyclase-activating peptide (PACAP) is a 38-amino acid polypeptide, first isolated from ovine hypothalamus, which directly stimulates the release of several pituitary hormones, including GH, ACTH, and LH. The presence of PACAP receptors in several brain areas, including the hypothalamus, suggests that this peptide might play a role as neurotransmitter/neuromodulator. We have thus investigated the effects of intracerebroventricular (i.c.v.) and intravenous (i.v.) injections of PACAP and the potent PACAP antagonist PACAP(6-38) on gonadotropin-releasing hormone (GnRH) and somatostatin (SS) gene expression in the male rat hypothalamus. The levels of mRNA were measured at the cellular level by quantitative in-situ hybridization. The i.c.v. injection of PACAP produced a 12.5% increase in the GnRH mRNA levels, an effect which was completely prevented by the concomitant administration of the PACAP antagonist. The administration of the PACAP antagonist induced by itself a 12.9% decrease in the hybridization signal. The i.v. administration of the same peptides induced modifications in GnRH gene expression which were completely opposite to those produced by i.c.v. administration. In somatostatinergic neurons located in the periventricular nucleus, the i.c.v. injection of the peptides induced modification in SS gene expression which were very similar to those observed for GnRH gene expression, although the changes were less striking. The i.v. administration of PACAP or its antagonist did not induce any change in the levels of SS mRNA. These results then strongly suggest that PACAP might be involved in a positive regulation of two neuropeptides involved in the control of anterior pituitary secretion via central specific receptors. The inverse influence of PACAP on GnRH gene expression after the i.v. injection might be explained by the short feedback effect induced by the direct stimulation of gonadotropin hormone release following the systemic injection of the peptide.