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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.

We studied the distribution of calretinin immunoreactivity (CR-ir) in the rostral and intermediate levels of the spinal cord of lampreys from embryonic to adult periods. CR-ir was first observed at hatching in motoneurons and primary sensory neurons of the spinal cord, the dorsal cells. During the prolarval period two new cell types showed CR-ir: ganglion cells and interneurons. Motoneurons, dorsal cells, and ganglion cells were strongly positive, whereas interneurons were weakly stained in late prolarvae. The intensity of CR-ir in these four types of cells changed during the larval period. Increase of CR-expression was found in interneurons but a decrease in dorsal cells and in ganglion cells. These changes were more evident in premetamorphic larvae. Postmetamorphic lampreys showed almost no CR-ir in dorsal cells. In adult lampreys, the interneurons showed the highest CR-ir, whereas motoneurons were more weakly stained than in earlier stages of development. Moreover, in adults the dorsal cells and the ganglion cells showed no CR-ir. The present study shows that CR-ir changes during lamprey spinal cord development in different types of neurons, sometimes in opposite ways. This plasticity of CR-expression may indicate different needs from subsets of lamprey spinal cord cells involved in the different locomotor behaviors along its life cycle.