Previous studies in anaesthetized animals have shown that the baroreflex control of sympathetic vasomotor activity is mediated to a large extent by inhibitory inputs to sympathoexcitatory pressor neurons in the rostral part of the ventrolateral medulla. The aim of this study was to determine, in conscious rabbits, the distribution of neurons within the brain that have two properties characteristic of interneurons conveying baroreceptor signals to the rostral ventrolateral medulla: (i) they are activated by an increase in arterial pressure; and (ii) they project specifically to the rostral ventrolateral medulla pressor region. In a preliminary operation, an injection of the retrogradely transported tracer, fluorescent-labelled microspheres, was made into the physiologically identified pressor region in the rostral ventrolateral medulla. After a waiting period of one to eight weeks, hypertension was produced in the conscious rabbit by continuous intravenous infusion of phenylephrine at a rate sufficient to increase arterial pressure by approximately 20 mmHg, maintained for a period of 60 min. A control group of animals was infused with the vehicle solution alone. In confirmation of our previous study,36 hypertension produced by phenylephrine resulted in the neuronal expression of Fos (a marker of neuronal activation) in the nucleus of the solitary tract, area postrema, the intermediate and caudal parts of the ventrolateral medulla parabrachial complex, and in the central nucleus of the amygdala. Approximately 50% of the Fos-immunoreactive neurons in both the caudal and intermediate parts of the ventrolateral medulla were also retrogradely labelled from the rostral ventrolateral medulla pressor region; such double-labelled neurons were confined to a discrete longitudinal column located just ventrolateral to the nucleus ambiguus. Significant numbers of double-labelled neurons were also found in the nucleus of the solitary tract and area postrema, although these represented a much lower proportion (13-16%) of the total number of Fos-immunoreactive neurons in these regions. In the parabrachial complex, Fos-immunoreactive and retrogradely labelled neurons were largely separate populations, while in the amygdala they were entirely separate populations. In the control group of rabbits, virtually no double-labelled neurons were found in any of these regions. The results indicate that putative baroreceptor interneurons that project to the pressor region of the rostral ventrolateral medulla are virtually confined to the lower brainstem. In particular, they support the results of previous studies in anaesthetized animals indicating that neurons in the intermediate and caudal ventrolateral medulla convey baroreceptor signals to the rostral ventrolateral medulla pressor region, and extend them by demonstrating the precise anatomical distribution of these neurons. Finally, the results demonstrate the existence of direct projections to the rostral ventrolateral medulla pressor region from the nucleus of the solitary tract and area postrema, and to a lesser extent the parabrachial complex, that may also play an important role in the baroreflex control of sympathetic vasomotor activity in conscious animals.