AbstractFeeding behavior is regulated by both the homeostatic needs of the body and hedonic values of the food. Easy access to palatable energy-dense foods and the consequent obesity epidemic stress the urgent need for a better understanding of neural circuits that regulate hedonic feeding. Here, we report that neurotensin-positive neurons in the lateral septum (LS^Nts) play a crucial role in regulating hedonic feeding. Silencing LS^Nts specifically promotes feeding of palatable food, whereas activation of LS^Nts suppresses overall feeding. LS^Nts neurons project to the tuberal nucleus (TU) via GABA signaling to regulate hedonic feeding, while the neurotensin signal from LS^Nts→the supramammillary nucleus (SUM) is sufficient to suppress overall feeding. In vivo calcium imaging and optogenetic manipulation reveal two populations of LS^Nts neurons that are activated and inhibited during feeding, which contribute to food seeking and consumption, respectively. Chronic activation of LS^Nts or LS^Nts→TU is sufficient to reduce high-fat diet-induced obesity. Our findings suggest that LS^Nts→TU is a key pathway in regulating hedonic feeding.