In mammals, glucosensing neurons reside in brain areas known to play a critical regulatory role in energy homeostasis and neuroendocrine function. In fish, although no glucosensing neurons have been characterized to date, glucokinase (GCK; the main glucosensing marker in mammals) activity and expression were found in hypothalamus and hindbrain of rainbow trout where they related to food intake regulation and glucose homeostasis. However, there are no available studies in literature regarding GCK distribution in brain areas as well as the cell types expressing that protein. In the present study we hypothesize that, as occurs in mammals, GCK would be localized where glucosensing areas have been described. In this sense, we have found GCK immunoreactivity in several areas of trout hypothalamus, of which some of them are related to glucose metabolism, energy homeostasis and food intake, including the lateral hypothalamus, anterior tuberal nucleus, posterior tuberal nucleus and lateral tuberal nucleus. On the other hand, GCKimmunoreactivity was also observed in other areas where the glucosensor system is probably functional,such as the preoptic area and the oculomotor nucleus. Therefore, in this study using immunoreactive techniques, we have demonstrated in those specific areas of the rainbow trout brain previously described as glucosensor the presence of GCK in different cell types.