Project Summary/Abstract Abnormalities in feeding behaviors are a key symptom of several conditions, including binge-eating disorder, anorexia and obesity. In order to develop novel treatments, it is imperative to gain a deeper understanding of the circuits controlling adaptive feeding. Eating is affected by metabolic and hedonic features, and it consists of several actions, including exploratory food-seeking and consumption. Despite its central clinical and biological importance, networks controlling these behaviors are not well-understood. Prior data have indicated that activation of several GABAergic inputs to the midbrain lateral and ventrolateral periaqueductal gray (l/vlPAG) elicit hunting of insects in mice. However, the role of local l/vlPAG GABAergic cells in feeding is unknown. Intriguingly, our preliminary data show that these cells encode food-seeking actions, such as approach to food and consumption. Furthermore, activity in these ensembles, or of their projections to the subthalamic zona incerta (ZI) are required for foraging leading to consumption of both prey and non-prey food. Here, we propose to combine converging advances in neural activity recording, computational methods and molecular circuit dissection tools to: 1. Characterize how l/vlPAG VGAT cells encode food sources and food-seeking behaviors by recording the neural activity of large ensembles of cells with miniaturized microscopes. 2. Determine if activity in the l/vlPAG circuit and their projection to the ZI is necessary and sufficient to promote foraging leading to consumption, and 3. Dissect how the l/vlPAG input affects the ZI by combining ex vivo and in vivo recordings of neural activity. Since prior reports show ZI activation induces feeding, we hypothesize that activation of the inhibitory l/vlPAG input to ZI elicits feeding by disinhibiting the ZI. Importantly, feasibility for all proposed aims is demonstrated in preliminary data and our prior publications, and we have successfully