Carnivorous fish have a limited capacity to utilize dietary carbohydrates even though glucosensing components exist in the hypothalamus and Brockmann bodies. Therefore, we fed trout for 10 days with two experimental diets containing a high level of carbohydrates (20%) or a carbohydrate-free level (<0.3%) to test the capacity of dietary carbohydrates to regulate gene expression of glucosensing components. Fish were fed and killed 1, 6, and 24 h after the meal to analyze plasma glucose levels, glucosensing-related biochemical parameters, and gene expression of the major components of the glucosensing system in the hypothalamus and Brockmann bodies. Glucose facilitative transporter type 2 and glucokinase gene expression were confirmed by real-time PCR data and two new components of the glucosensing mechanism, Kir6.-like and sulfonylurea receptor-like, were detected for the first time in fish in both tissues. In addition, a clear adaptation to dietary carbohydrates was found in trout Brockmann bodies, based on increased gene expression of major components of the system as well as enhanced glucokinase activities and glycogen levels. In contrast, in the hypothalamus, only glucokinase gene expression and activity showed a response to dietary carbohydrates, supporting the key role of that enzyme in glucosensing mechanism. Finally, a differential postprandial profile was found between tissues regarding the glucosensing potential, since the hypothalamus seems to respond to hyperglycemia earlier than the Brockmann bodies, whose response took place later. Altogether, these data describe for the first time in fish a complete response of major glucosensing components to dietary carbohydrates in trout hypothalamus and Brockmann bodies, supporting an efficient adaptation of both tissues to those dietary components.