Hypotension produces a reflex increase in the activity of sympathetic vasomotor nerves. Studies in anaesthetised animals have established that neurons in the rostral ventrolateral medulla (RVLM) that project directly to sympathetic vasomotor preganglionic neurons in the spinal cord are a critical component of the central pathways mediating this reflex response. There are also neurons in supramedullary regions (the A5 area in the pons and the paraventricular nucleus (PVN) in the hypothalamus), however, that project directly to the sympathetic vasomotor outflow. The aim of this study was to identify and map neurons within the A5 area and PVN, as well as in the RVLM, which may contribute to the reflex sympathoexcitatory response to a hypotensive challenge in conscious rabbits. In a preliminary operation, a retrogradely transported tracer was injected into a site centred on the intermediolateral cell column in the upper lumbar spinal cord. After a waiting period of at least 1 week, a moderate hypotension (decrease in arterial pressure of approximately 20 mm Hg) was induced in conscious rabbits for 60 min by continuous infusion of sodium nitroprusside. In confirmation of previous studies, hypotension resulted in the expression of Fos in the RVLM, the A5 area and PVN. There were also retrogradely labelled neurons in all these regions. In both the RVLM and A5 area, approximately 40% of the retrogradely labelled neurons were also immunoreactive for Fos. In contrast, in the PVN the proportion of retrogradely labelled neurons that were also Fos-positive was much less (approximately 6%). This study has demonstrated that, in the conscious rabbit, a significant proportion of spinally projecting neurons within discrete regions in the RVLM and A5 area are activated by hypotension (as indicated by Fos expression). In the PVN, only a very small proportion of spinally projecting neurons are activated by hypotension, and thus these neurons appear to be regulated primarily by inputs other than baroreceptor inputs.