Summary: Twenty-four hour rhythms, at 4 h intervals, of norepinephrine (NE) and serotonin (5-HT) contents were investigated in the rat brain regions where sleep-wakefulness regulation is believed to occur: Nucleus suprachiasmaticus (SC), n, raphe dorsalis (RD) and medialis (RM), and locus coeruleus, Cosinor method of Halberg was applied to evaluate sinusoidal rhythmicity of the measured values, In the SC only NE showed a significant rhythm with a peak value at the beginning of the light period, which suggests that a NE mechanism may be involved in oscillating biological rhythms in rats, In the RD and RM, 5-HT and 5-hydroxyindoleacetic acid increased significantly during the light period, Moreover, 5-HT rhythm in the RD was maintained even under constant dark conditions, which suggests that 5-HT rhythm in the RD may be endogenous, Key Words: Twentyfour hour rhythm-Norepinephrine-Serotonin-Nucleus suprachiasmaticusRaphe nuclei-Locus coeruleus, Most physiological functions in rodents, such as locomotion, feeding and drinking, thermoregulation, and hormonal secretion, exhibit circadian rhythms. These rhythms are believed to be controlled by a pacemaker located in the nucleus suprachiasmaticus (SC) of the hypothalamus (1). Physiological rhythms may be affected by biochemical changes in the brain. It has been demonstrated that norepinephrine (NE) and serotonin (5-HT) contents in the whole brain or some brain regions change over a 24 h period (2-6). Recently Ibuka et al. (7) found that the sleep-wakefulness cycle is also regulated by the circadian pacemaker in the SC. Although Jouvet has recently modified his monoamine hypothesis of sleep and stressed the importance of the cholinergic mechanisms (8,9), studying the interactions between 5-HT and NE in sleep-wakefulness regulation is still useful.We have studied the 24 h changes in 5-HT and NE contents in the n. raphe dorsalis (RD) and medialis (RM), locus coeruleus (LC), and SC in the rat. To investigate whether a 24 h rhythm in the raphe nuclei is endogenous, an additional experiment was carried out.