Orexins A and B are hypothalamic peptides derived from a precursor called prepro-orexin and are associated with the stimulation of food intake and arousal. There is evidence that orexins act on some pituitary functions. Since no studies have been done concerning the presence of orexins in human pituitary, it is unclear whether the local effect of these peptides is due to orexins synthesized in the pituitary or to circulating-derived orexins. To define a possible paracrine regulatory role of orexins on pituitary cell function, we have sought to characterize the expression of orexins in the human adenohypophysis as well as to identify the cell types that express these proteins. In the present study, we used immunohistochemistry and double labeling to detect the presence of orexin A and orexin B in human pituitary. Orexin A was localized in 33% of pituitary cells. With double immunofluorescence techniques we demonstrated that orexin A is present in PRL (82 +/- 5.3%), TSH (18 +/- 2.3%), GH (10 +/- 2.3%), FSH (8 +/- 2.6%), and LH (7 +/- 3.2%) cells, but not in corticotroph cells. Orexin B was found in virtually all corticotrophs cells of the anterior pituitary. These results demonstrate that lactotroph cells are the main source of orexin A and corticotroph cells of orexin B. In summary the present findings provide the first evidence that orexins A and B are expressed in specific human pituitary cell types. Our data provide the cellular basis for a paracrine role of orexins in human pituitary cell function and further our understanding regarding the mechanisms by which orexins influence neuroendocrine function.