Plasma glucose concentrations display a daily rhythm generated by the hypothalamic biological clock, located in the suprachiasmatic nucleus (SCN). How the SCN orchestrates this rhythm is unknown. Because glucagon stimulates hepatic glucose production, we hypothesized that if glucagon has a daily rhythm, then it may be responsible for the glucose rhythm. From hourly blood samples, we determined daily glucagon concentrations for intact and SCN-lesioned rats. Intact ad libitum-fed rats showed a clear daily glucagon rhythm, and fasting resulted in an even more pronounced rhythm. It is interesting that a decrease in glucagon concentrations, instead of the expected increase, occurred already shortly after food removal. Toward the start of the active period, a peak in glucagon levels occurred, with concentrations similar to those measured in ad libitumfed rats. SCN lesions abolished rhythmicity in plasma glucagon profiles. Scheduled-fed rats showed meal-induced glucagon peaks but also a daily rhythm in basal premeal glucagon concentrations. Plasma glucose concentrations of ad libitum-and scheduled-fed rats, however, were similar. In conclusion, feeding and the biological clock control 24-h plasma glucagon concentrations. In fed rats, glucagon is not responsible for the daily glucose rhythm. During fasting, however, glucagon may contribute to energy mobilization when the activity period starts. Diabetes 52: 1709 -1715, 2003 M ammals adapt their activity pattern to the daily changes in light intensity. This activity rhythm dictates the need for energy at a specific time of day. This includes glucose in particular, because under normal conditions, it is the only fuel that can be metabolized by the brain (1). Circadian rhythms have been shown in mammals (2,3), including humans (4,5). Rat plasma glucose concentrations display a daily rhythm, with peak values at the beginning of the activity period. Studies involving scheduled feeding and thermic lesions of the biological clock, located in the suprachiasmatic nucleus (SCN), have shown that this rhythm is feeding independent and generated by the SCN (6,7), yet it is not clear by which mechanism the SCN creates this glucose rhythm. Studies investigating the role of insulin have shown that insulin concentrations have a daily rhythm but not independent of food intake (6) and therefore cannot be responsible for the glucose rhythm. Corticosterone stimulates gluconeogenesis (8) and also displays a clear daily rhythm (9 -12), with peak values occurring just before the onset of the activity period. Despite the hyperglycemic effect of corticosterone and the coincidence of its peak release with the peak in plasma glucose concentrations, blocking corticosterone synthesis does not affect the morning rise of glucose concentrations seen in humans (13) and thus cannot be the main cause of the rhythm in glucose concentrations. Growth hormone, too, is able to stimulate glucose release (8) but in the rat displays an ultradian rhythm rather than a circadian rhythm, which makes it unlikely to...