The effect of noxious stimulation on the expression of FOS-like immunoreactivity (FOS-LI) in neurons of the parabrachial nucleus (PB) was studied in awake, freely moving rats. In one series of experiments, the rats were subjected to noxious mechanical stimulation (pinch) of either the nape of the neck or the base of the tail for 20 seconds every 5 minutes for 90 minutes, and then they were killed by transcardial perfusion after 45-210 minutes. Control animals received innocuous mechanical stimulation (brush) of the tail. Noxious stimuli resulted in FOS-LI in neurons in the dorsal part of the lateral PB, with heavy labeling in the superior lateral (PBsl) and the dorsal lateral (PBdl) subnuclei. FOS-LI was also elicited in the central lateral subnucleus (PBcl) and, although much more sparsely, in the external lateral subnucleus and the Kölliker-Fuse nucleus. Tail and neck stimulation resulted in similar labeling patterns, but more neurons, particularly in PBsl, expressed FOS-LI after pinch of the tail than of the neck. In another series of experiments, rats received injection of 5% formalin into one hindpaw. After 75-90 minutes, FOS-LI was seen in the same parts of PB as after noxious mechanical stimulation. The heaviest labeling was seen on the side contralateral to the injection side, with statistically significant (P < 0.05) side differences present in PBsl and PBdl. In a third series of experiments, rats were hemisected at low cervical-upper thoracic segments, allowed 2 weeks to recover, and then given formalin injections in both hindpaws. Significantly more neurons were FOS-labeled in PBdl, PBsl, and PBcl on the side contralateral to the hemisection than on the ipsilateral side. These observations are discussed in relation to the organization of the spinal afferent input and the efferent connections of PB. It is concluded that the FOS-LI expression in PBdl and PBsl and probably also in PBcl, to a large extent, is evoked by the ascending spinal nociceptive input to PB. Because these subnuclei project to several hypothalamic regions, it is suggested that neurons in PB that express FOS after noxious mechanical and chemical stimulation primarily are involved in autonomic and homeostatic responses to behavioral situations that involve tissue-damaging stimuli.