Selective Fos induction in hypothalamic orexin/hypocretin, but not melanin-concentrating hormone neurons, by a learned food-cue that stimulates feeding in sated rats

Associative learning can enable cues from the environment to stimulate feeding in the absence of physiological hunger. How learned cues are integrated with the homeostatic regulatory system is unknown. Here we examined whether the underlying mechanism involves the hypothalamic orexigenic neuropeptide regulators orexin/hypocretin (ORX) and melanin-concentrating hormone (MCH). We used a Pavlovian conditioning procedure to train food-restricted rats to associate a discrete cue, a tone, with food pellets distinct from their regular lab chow diet. Rats in the conditioned group (Paired) received presentations of a tone immediately prior to food delivery, while the rats in the control group (Unpaired) received random presentations of the same number of tones and food pellets. After conditioning rats were allowed ad libitum access to lab chow for at least 10 days before testing. At test sated rats were presented with the tones in their home cages, and then one group was allowed to consume food pellets, while another group was left undisturbed until sacrifice for Fos induction analysis. The tone cue stimulated food consumption in this setting; rats in the Paired group consumed larger amounts of food pellets than rats in the Unpaired group. To examine Fos induction we processed the brain tissue using fluorescent immunohistochemistry methods for combined detection of Fos and characterization of ORX and MCH neurons. We found a greater percentage of ORX and Fos double-labeled neurons in the Paired compared to the Unpaired condition, specifically in the perifornical area. In contrast, there were very few MCH neurons with Fos induction in both the Paired and Unpaired conditions. Thus, the food-cue selectively induced Fos in ORX but not in MCH neurons. These results suggest a role for ORX in cue-induced feeding that occurs in the absence of physiological hunger.

► Learned food-cues can stimulate feeding in the absence of physiological hunger. ► How food-cues are integrated with the homeostatic physiological control is unknown. ► We examined Fos induction in lateral hypothalamic orexigenic neuropeptide neurons. ► Food-cue induced Fos in orexin but not in melanin-concentrating hormone neurons.