In response to a pain-related acute stress, the expression of c-fos protein (Fos), a marker of acute neuronal excitation, was investigated in the hypothalamus of rats. Few Fos-immunopositive cells were seen 15 min after a single subcutaneous injection of 4% formalin in the hypothalamus, but only in the paraventricular nucleus (PVN). Fifteen minutes later, a high number of parvocellular neurons of the PVN showed Fos expression. By 60 min after injection, strong immunoreactivity appeared in the arcuate nucleus, but the Fos-positive neurons distributed almost exclusively in the ventromedial subdivision of the nucleus. Neurons in this part of the arcuate nucleus express mainly neuropeptide Y (NPY) that projects to the medial parvocellular subdivision of the PVN. It has been demonstrated by previous studies that this part of the arcuate nucleus receives blood partly from the anterior pituitary through the subependymal plexus of the median eminence, and that it establishes, together with the median eminence, a blood-brain barrier-free area in the medial basal hypothalamus. Since the PVN-projecting NPY neurons in the arcuate neurons are sensitive to alterations in circulating corticosterone levels, the existence of a possible short feedback route in the stress-activated hypothalamo-pituitary-adrenocortical system is discussed.