By means of two‐colour immunocytochemistry using a mouse monoclonal antibody directed against the rat liver glucocorticoid receptor (GR) and a rabbit polyclonal neuropeptide Y (NPY) antiserum combined with the biotin‐avidin immunoperoxidase and a double immunofluorescence procedure, it has been possible to demonstrate nuclear GR immunoreactivity (IR) in neurons showing cytoplasmatic NPY IR in rat brain. The majority of NPY immunoreactive perikarya of the medial parvocellular part of the arcuate nucleus, locus coeruleus and the rostral and caudal part of the ventrolateral medulla oblongata contained strong nuclear GR IR. Many of the NPY immunoreactive neurons present in the subnuclei of the nucleus tractus solitarius also contained nuclear GR IR, while most of the NPY immunoreactive perikarya of the cerebral cortex and all of the neostriatum appeared to lack GR IR. These results indicate that NPY immunoreactive neurons in the upper and lower brain stem, but not in the cerebral cortex and in the neostriatum may be directly involved in mediating central effects of glucocorticoids.