Heterotrimeric G-proteins, comprising alpha, beta and gamma subunits, have been shown to play a central role in coupling multiple receptors to a variety of enzymes and ion channels. In vitro studies have demonstrated the existence of selective interactions between various alpha, beta and gamma subunits, as well as between specific heterotrimers and target receptor and effector proteins. However, little is known of the physiological relevance of such associations, and the determinants of specificity in G-protein signaling within the brain remain largely unidentified. To investigate the possibility that specific heterotrimeric interactions result from discrete localizations of the G-protein subunits within the brain, we have used the technique of in situ hybridization to map the distribution of G-protein beta and gamma subunits in the rat brain. Beta1, beta2, beta3 and beta5 subunits were found to be widely expressed throughout the rat brain, whilst beta4 and the G-protein gamma subunit messenger RNAs generally showed more discrete expression patterns. The expression patterns for these subunits suggest that individual beta and gamma subunits may be co-expressed in certain cell types and brain regions; a particularly intriguing and striking co-localization was observed in the case of beta4 and gamma2 subunit messenger RNAs in layer VI of the occipital cortex. The localizations of the G-protein beta and gamma subunits, and their potential coupling to various receptor/effector systems, are discussed.