||OBJECTIVE: Intrinsically photosensitive retinal ganglion cells are most sensitive to short wavelengths and reach brain regions that modulate biological rhythms and energy metabolism. The increased exposure nowadays to artificial light at night (ALAN), especially short wavelengths, perturbs our synchronization with the 24-hour solar cycle. Here, the time- and wavelength dependence of the metabolic effects of ALAN are investigated. METHODS: Male Wistar rats were exposed to white, blue, or green light at different time points during the dark phase. Locomotor activity, energy expenditure, respiratory exchange ratio (RER), and food intake were recorded. Brains, livers, and blood were collected. RESULTS: All wavelengths decreased locomotor activity regardless of time of exposure, but changes in energy expenditure were dependent on the time of exposure. Blue and green light reduced RER at Zeitgeber time 16-18 without changing food intake. Blue light increased period 1 (Per1) gene expression in the liver, while green and white light increased Per2. Blue light decreased plasma glucose and phosphoenolpyruvate carboxykinase (Pepck) expression in the liver. All wavelengths increased c-Fos activity in the suprachiasmatic nucleus, but blue and green light decreased c-Fos activity in the paraventricular nucleus. CONCLUSIONS: ALAN affects locomotor activity, energy expenditure, RER, hypothalamic c-Fos expression, and expression of clock and metabolic genes in the liver depending on the time of day and wavelength.