Neuropeptide Y (NPY) is colocalized with catecholamines in brainstem-hypothalamus neuronal pathways implicated in the regulation of the hypothalamo-hypophyso-adrenocortical system (HHAS) and may interact with catecholamines in the regulation of neuroendocrine stress responses. On the other hand, the presence of corticosteroid receptors in these neurons suggests that adrenocortical hormones may regulate central NPY levels. NPY and biogenic amine levels were therefore measured in punched brain regions (ventrolateral and dorsomedial medulla, paraventricular and arcuate nucleus of the hypothalamus, frontal cortex) in basal conditions and up to 4 h after a 30-min period of restraint stress. Despite a massive activation of the HHAS (measured by plasma levels of ACTH and glucocorticoids), and a large increase of prolactin levels, no change of NPY levels could be found in any of the above-mentioned structures. The same parameters, as well as plasma levels of NPY and catecholamines and NPY levels in cardiac tissue were also measured 2, 20 and 96 h after adrenalectomy. Again, despite the large changes of brain and peripheral catecholamine levels, and the intense activation of the HHAS, no change of NPY levels was observed in any of the structures studied. These data question the nature of the relationships between NPY and neuroendocrine stress responses.