Gastrin-releasing peptide (GRP) is a 27 amino acid peptide that is present in both the central and peripheral nervous systems and that shares immunological and functional properties with the amphibian peptide, bombesin. GRP has multiple putative biological functions including effects on feeding behaviour and carbohydrate metabolism, body temperature, and effects on hormone release, but little is known about the regulation of GRP gene expression in the brain. This study examined the distribution of neurones expressing preproGRP mRNA in rat brain by in situ hybridization of [35S]-labelled DNA oligonucleotides. PreproGRP mRNA was detected in several regions of brain, with highest concentrations in the parvocellular paraventricular and suprachiasmatic nuclei of the hypothalamus, the lateral and basolateral nuclei of the amygdala, the amygdaloid-hippocampal area and the ventral part of the granule cell layer of the dentate gyrus. Moderate levels were seen in layers II and III of the cingulate and retrosplenial cortex, the medial and mediobasal nuclei of the amygdala, the anteroventral thalamic nucleus; medial geniculate nucleus and the parabrachial nucleus. These findings are largely consistent with the cellular localization of GRP-like immunoreactivity in rat brain and recent studies of preproGRP mRNA localization using cRNA probes. The distribution of preproGRP mRNA observed further suggests the involvement of GRP in the central regulation of several functions including regulation of hypothalamic/pituitary hormone release.