We have mapped Fos expression to investigate brain regions activated by synthetic selective I-2 (BU224) and endogenous (harmane) imidazoline binding site ligands in naive and restraint-stressed rats. Systemic administration of BU224 or harmane to naive rats increased Fos-like immunoreactivity (FLI) in the hypothalamic paraventricular nucleus (PVN), hippocampal dentate gyrus (DC), central and medial. nuclei of the amygdala (CeA, MeA) and the locus coeruleus (LC). FLI in restraint-stressed rats was increased in all 5 regions by harmane, and in the CeA MeA and, LC by BU224. Dual-labelling of FLI cells in the PVN of naive rats showed an increase in the number of corticotrophin-releasing-factor-containing cells (CRF) activated by BU224 and harmane. Several CRF-containing neurons in the PVN expressed alpha(1)-adrenoceptors and were densely surrounded by catecholaminergic axons and terminals. Our results provide a functional neuroanatomical framework which may explain the stimulatory effects of imidazoline ligands on basal and stress-induced neuronal and neuroendocrine activity. (C) 2009 Elsevier B.V. and ECNP. All rights reserved.