When food availability is restricted to a particular time each day, mammals exhibit food-anticipatory activity (FAA), a daily increase in locomotor activity preceding the presentation of food. Considerable historical evidence suggests that FAA is driven by a foodentrainable circadian clock distinct from the master clock of the suprachiasmatic nucleus. Multiple food-entrainable circadian clocks have been discovered in the brain and periphery, raising strong expectations that one or more underlie FAA. We report here that mutant mice lacking known circadian clock function in all tissues exhibit normal FAA both in a light-dark cycle and in constant darkness, regardless of whether the mutation disables the positive or negative limb of the clock feedback mechanism. FAA is thus independent of the known circadian clock. Our results indicate either that FAA is not the output of an oscillator or that it is the output of a circadian oscillator different from known circadian clocks.food-anticipatory activity ͉ mouse genetics W hen food availability is limited to a several-hour interval at a particular time each day, mammals quickly develop a new component of daily behavioral activity, a second period of arousal and increased locomotor activity that occurs shortly before the time of daily food presentation (1). This so-called food-anticipatory activity (FAA) is robust, stable over many daily cycles, and occurs even if the time of food availability lies within the light phase of the light-dark cycle, the resting period of the daily behavioral cycle for a nocturnal laboratory animal. Because feeding strategies are fundamental to survival, it is thought that FAA represents an evolutionary adaptation providing mammals with a highly flexible and efficient food-seeking program, one that is able, if necessary, to take advantage of food sources available at unusual times with respect to the animal's typical daily rest-activity cycle (2).The mechanisms underlying FAA have been under investigation for nearly 30 years. The accumulated evidence suggests that FAA is more likely to be generated by a food-entrainable circadian oscillator than by plausible alternatives, such as a passive hourglass mechanism or an associative memory process (1). In general, an unequivocal demonstration that a rhythmic process is driven by an oscillator requires observation of the persistence of the rhythm over multiple cycles following removal of the entraining stimulus. In the case of daily rhythms entrained to a light-dark cycle, testing for a postulated underlying circadian oscillator simply requires switching off the lights and monitoring the daily rhythms for persistence in constant darkness. For example, daily rhythms of locomotor activity, feeding, and drinking in a light-dark cycle persist indefinitely after a transition to constant darkness, and were thus long ago demonstrated to be driven by a light-entrainable circadian oscillator (3). In contrast, a compelling demonstration that FAA relies on a self-sustained, food-entrainable oscillator has been...