The serotonergic system plays a key role in the regulation of brain states, and many of the known features of serotonergic neurons appear to match this function. Midbrain raphe nuclei provide a diffuse projection to all regions of the forebrain, and raphe neurons exhibit a slow metronome-like activity that sets the ambient levels of serotonin across the sleep-wake cycle. Serotonergic cells have also been implicated, however, in a variety of more specific functions that can hardly be related to their low-rate monotonous patterns of discharges. The amazing variety of serotonergic receptors and their type-specific distribution on cortical neurons also raise the possibility of a more intimate coordination between the activity of serotonergic neurons and their target cortical circuits. Here we report an unexpected diversity in the behavior of immunohistochemically identified serotonergic neurons. Two outstanding subpopulations were identified by using the in vivo juxtacellular recording and labeling technique. The first subpopulation of serotonergic cells exhibited the classic clock-like activity with no apparent short timescale interaction with the hippocampal electroencephalogram. The other subpopulation discharged action potentials that were phase-locked to the hippocampal theta rhythm, the oscillatory pattern associated with acquisition of information and memory formation. These results indicate that the ascending serotonergic system comprises cells involved in complex information processing beyond the regulation of state transitions. The heterogeneity of serotonergic neuron behavior can also help to explain the complexity of symptoms associated with serotonergic dysfunction.hippocampus ͉ juxtacellular labeling ͉ midbrain raphe ͉ neuronal oscillations T he serotonergic system is a primary target of psychotherapy that stresses the importance of serotonin in normal and abnormal brain functioning. Serotonergic neurons of the midbrain raphe have been implicated in the control of affective and cognitive functions and in modulating the neural activities of networks across the sleep-wake cycle. Behaviorally related oscillations in the theta frequency range (4-8 Hz), which are most prominent in the hippocampus (1), are known to synchronize neuronal activity in a number of forebrain structures involved in these functions (2-4). Hippocampal theta oscillations occur selectively during exploratory behaviors and paradoxical sleep, whereas large-amplitude irregular activity is associated with quiet waking, consummatory behaviors, and slow-wave sleep (5, 6). As in sleep, hippocampal activity spontaneously alternates between theta and nontheta states under urethane anesthesia.The majority of putative serotonergic neurons have been reported to be most active during waking, to decrease their firing rate during slow-wave sleep, and to cease firing during paradoxical sleep (7). ʈ The silencing of serotonergic cells during paradoxical sleep is currently considered an important regulatory signal that contributes to the maintenance of ...