1. This study was designed to assess the molecular and cellular events involved in the up-regulation (and receptor supersensitivity) of brain alpha(2)-adrenoceptors as a result of chronic depletion of noradrenaline (and other monoamines) by reserpine. 2. Chronic reserpine (0.25 mg kg(-1) s.c., every 48 h for 6 - 14 days) increased significantly the density (B(max) values) of cortical alpha(2)-adrenoceptor agonist sites (34 - 48% for [(3)H]-UK14304, 22 - 32% for [(3)H]-clonidine) but not that of antagonist sites (11 - 18% for [(3)H]-RX821002). Competition of [(3)H]-RX821002 binding by (-)-adrenaline further indicated that chronic reserpine was associated with up-regulation of the high-affinity state of alpha(2)-adrenoceptors. 3. In cortical membranes of reserpine-treated rats (0.25 mg kg(-1) s.c., every 48 h for 20 days), the immunoreactivities of various G proteins (Galphai(1/2), Galphai(3), Galphao and Galphas) were increased (25 - 34%). Because the high-affinity conformation of the alpha(2)-adrenoceptor is most probably related to the complex with Galphai(2) proteins, these results suggested an increase in signal transduction through alpha(2)-adrenoceptors (and other monoamine receptors) induced by chronic reserpine. 4. After alpha(2)-adrenoceptor alkylation, the analysis of receptor recovery (B(max) for [(3)H]-UK14304) indicated that the increased density of cortical alpha(2)-adrenoceptors in reserpine-treated rats was probably due to a higher appearance rate constant of the receptor ((Delta)r=57%) and not to a decreased disappearance rate constant ((Delta)k=7%). 5. Northern- and dot-blot analyses of RNA extracted from the cerebral cortex of saline- and reserpine-treated rats (0.25 mg kg(-1), s.c., every 48 h for 20 days) revealed that reserpine markedly increased the expression of alpha(2a)-adrenoceptor mRNA in the brain (125%). This transcriptional activation of the receptor gene expression appears to be the cellular mechanism by which reserpine induces up-regulation in the density of brain alpha(2)-adrenoceptors.