Alternate day, intermittent fasting (IMF) can promote weight loss in obese individuals. We investigated the metabolic and neural mechanisms of IMF in diet-induced obese mice. Twenty-four C57 male mice at PND 49 were fed a high-fat diet (HFD; 45% fat, 20% protein, 35% CHO) ad libitum for 8 wks. Following this, for 4 wks, mice (n = 8/group) were maintained on ad libitum HFD, received IMF of HFD (IMF-HFD), or were switched to ad libitum low-fat diet (LFD;10% fat, 20% protein, 70% CHO). Body weight and cumulative calorie intake were not different between the IMF-HFD and LFD. Indeed, both groups had reduced cumulative caloric intake (~13% for both; p < 0.01) and reduced body weight (~10% for both; p < 0.05) compared with the HFD. Metabolic parameters, however, were different between the IMF-HFD and LFD groups. Insulin tolerance was improved in IMF-HFD compared with the HFD group (- 20% glucose AUC; p < 0.05), but impaired compared with LFD (+ 22% glucose AUC; p