To gain new knowledge on the CNS-dependent regulation of glucose homeostasis, we employed an unbiased experimental approach to profile molecular signatures that define the neuronal populations activated after the ingestion of a hypercaloric high-sucrose diet (HSD). The overarching goal is to gain deeper insights into the neuronal regulation of systemic glucose homeostasis and insulin sensitivity during caloric overload, as a first step toward developing new therapies to treat diabetes—a disease that is rapidly becoming a major health problem in Europe and beyond, with concurrent negative impacts on quality of life as well as public health costs.