Long term effects of sleep deprivation on the mammalian circadian pacemaker

STUDY OBJECTIVES: In mammals, sleep is controlled by a homeostatic process, which regulates depth of sleep, and by the circadian clock of the suprachiasmatic nucleus (SCN), which regulates 24-h rhythms in timing of sleep. Sleep deprivation is known to cause molecular and physiological changes and results in an alteration in the timing of sleep. It is generally assumed that following sleep deprivation, homeostatic mechanisms overrule the circadian clock, allowing animals to sleep during their active phase. However, recent evidence indicates that sleep states have direct access to the circadian pacemaker of the SCN. We questioned therefore whether sleep deprivation may have long-term effects on the circadian pacemaker, which may explain altered sleep patterns following sleep deprivation. DESIGN: To test this hypothesis, we combined SCN recordings of electrical impulse frequency through stationary implanted electrodes in freely moving rats with electroencephalogram recordings in the same animal before, during, and after a mild 6-h sleep deprivation. MEASUREMENTS AND RESULTS: Following sleep deprivation, SCN neuronal activity was significantly reduced to about 60% of baseline levels. The decrements in SCN activity were most obvious during NREM sleep and REM sleep and lasted for 6-7 hours. CONCLUSIONS: The data show that sleep deprivation influences not only sleep homeostatic mechanisms, but also SCN electrical activity, resulting in a strong reduction in circadian amplitude in the major output signal from the SCN.     

Electrical synapses coordinate activity in the suprachiasmatic nucleus

In the suprachiasmatic nucleus (SCN), the master circadian pacemaker, neurons show circadian variations in firing frequency. There is also considerable synchrony of spiking across SCN neurons on a scale of milliseconds, but the mechanisms are poorly understood. Using paired whole-cell recordings, we have found that many neurons in the rat SCN communicate via electrical synapses. Spontaneous spiking was often synchronized in pairs of electrically coupled neurons, and the degree of this synchrony could be predicted from the magnitude of coupling. In wild-type mice, as in rats, the SCN contained electrical synapses, but electrical synapses were absent in connexin36-knockout mice. The knockout mice also showed dampened circadian activity rhythms and a delayed onset of activity during transition to constant darkness. We suggest that electrical synapses in the SCN help to synchronize its spiking activity, and that such synchrony is necessary for normal circadian behavior.     

The suprachiasmatic nucleus regulates sleep timing and amount in mice

CONTEXT: Sleep is regulated by circadian and homeostatic processes. The circadian pacemaker, located in the suprachiasmatic nuclei (SCN), regulates the timing and consolidation of the sleep-wake cycle, while a homeostatic mechanism governs the accumulation of sleep debt and sleep recovery. Recent studies using mice with deletions or mutations of circadian genes show that components of the circadian pacemaker can influence the total amount of baseline sleep and recovery from sleep deprivation, indicating a broader role for the SCN in sleep regulation. OBJECTIVE: To further investigate the role of the circadian pacemaker in sleep regulation in mice, we recorded sleep in sham and SCN-lesioned mice under baseline conditions and following sleep deprivation. RESULTS: Compared to sham controls, SCN-lesioned mice exhibited a decrease in sleep consolidation and a decrease in wakefulness during the dark phase. Following sleep deprivation, SCN-lesioned mice exhibited an attenuated increase in non-rapid eye movement sleep time but an increase in non-rapid eye movement sleep electroencephalographic delta power that was similar to that of the sham controls. CONCLUSIONS: These findings support the hypothesis that the SCN consolidate the sleep-wake cycle by generating a signal of arousal during the subjective night (ie. the active period), thereby having the capacity to alter baseline sleep amount. Although the SCN are not involved in sleep homeostasis as defined by the increase in electroencephalographic delta power after sleep deprivation, the SCN does play a central role in the regulation of sleep and wakefulness beyond just the timing of vigilance states.     

Fear conditioning-induced plasticity in auditory thalamus and cortex: To what extent is it expressed during slow-wave sleep?

After fear conditioning to a tone, rats received nonawakening presentations of the tone alone during slow-wave sleep (SWS) episodes. Multiunit activity was recorded in the medial part of the medial geniculate (MGm) and in the primary auditory cortex (ACx). Although tone-evoked responses were increased in MGm and ACx during the 3 conditioning sessions, group data failed to show any significant changes during SWS. Nonetheless, the few recordings (5/29) that exhibited the strongest conditioned responses during wakefulness expressed enhanced responding during SWS. Compared with previous data obtained in MGm during paradoxical sleep, associative plastic changes were less easily expressed during SWS. These results are discussed with regard to functional changes that occur in the thalamocortical system across vigilance states.     

Impaired sodium levels in the suprachiasmatic nucleus are associated with the formation of cardiovascular deficiency in sleep-deprived rats

Biological rhythms are a ubiquitous feature of all higher organisms. The rhythmic center of mammals is located in the suprachiasmatic nucleus (SCN), which projects to a number of brainstem centers to exert diurnal control over many physiological processes, including cardiovascular regulation. Total sleep deprivation (TSD) is a harmful condition known to impair cardiovascular activity, but the molecular mechanisms are unknown. As the inward sodium current has long been suggested as playing an important role in driving the spontaneous firing of the SCN, the present study aimed to determine if changes in sodium expression, together with its molecular machinery (Na-K ATPase) and rhythmic activity within the SCN, would occur during TSD. Adult rats subjected to different periods of TSD were processed for time-of-flight secondary ion mass spectrometry, Na-K ATPase assay, and cytochrome oxidase (COX) (an endogenous bioenergetic marker for neuronal activity) histochemistry. Cardiovascular dysfunction was determined through analysis of heart rate and changes in mean arterial pressure. Results indicated that, in normal rats, strong sodium signals were expressed throughout the entire SCN. Enzymatic data corresponded well with spectrometric findings in which high levels of Na-K ATPase and COX were observed in this nucleus. However, following TSD, all parameters including sodium imaging, sodium intensity as well as COX activities were drastically decreased. Na-K ATPase showed an increase in responsiveness following TSD. Both heart rate and mean arterial pressure measurements indicated an exaggerated pressor effect following TSD treatment. As proper sodium levels are essential for SCN activation, reduced SCN sodium levels may interrupt the oscillatory control, which could serve as the underlying mechanism for the initiation or development of TSD-related cardiovascular deficiency.     

The effects of continuous light exposure on Syrian hamster suprachiasmatic (SCN) neuronal discharge activity in vitro

The circadian variation in neuronal discharge activity recorded in vitro from the Syrian hamster suprachiasmatic nucleus (SCN), identified as a circadian clock, was used as an index of SCN circadian function. SCN neurones (n = 89) following in vitro commissural section of the paired SCN showed a similar peak in spontaneous discharge activity during the projected light phase (between CT 06.00 and 08.00 h) as the commissural intact LD 12:12 preparations (n = 230 neurones). Long-term exposure to continuous lighting (LL) may induce either arrhythmicity or splitting of locomotor behaviour. Circadian variation in SCN discharge activity was absent in hamsters (44 cells, 6 animals) showing LL-induced arrhythmicity. In 4 hamsters exhibiting split free-running behaviour two peaks in SCN discharge activity (n = 32 cells) were observed. Fifteen LL free-running animals showed no evidence of splitting or arrhythmicity and subsequent SCN recordings revealed only a single peak in SCN discharge activity. These LL-induced changes in overt circadian behaviour appeared paralleled by changes in neuronal discharge activity of the SCN circadian clock and support the view that the paired SCN act as a mutually coupled circadian oscillator.     

Human thalamic and cortical activities assessed by dimension of activation and spectral edge frequency during sleep wake cycles

STUDY OBJECTIVES: Using spectral edge frequency (SEF95) and dimension of activation (DA), a new tool derived from the dimension of correlation, we assessed the activation of thalamus and cortex in the different vigilance states. PATIENTS: Results were gathered from intracerebral recordings performed in 12 drug-resistant epileptic patients during video-stereoelectroencephalographic (SEEG) monitoring. RESULTS: In the cortex, we observed a progressive decrease of DA from wake to sleep, with minimal DA values characterizing the deep slow wave sleep (dSWS) stage. During paradoxical sleep (PS), cortical level of activity returned to DA values similar to those obtained during wakefulness. In the thalamus, DA values during wakefulness were higher than the values observed during light SWS (ISWS), deep SWS (dSWS) and PS; there were no significant differences between the 3 sleep stages. Similar variations were observed with SEF95. CONCLUSION: DA analysis proved reliable for quantification of cortical activity, in agreement with data issued from classical vigilance states scoring and spectral analysis. At the thalamic level, only 2 levels of activity within a sleep wake cycle were observed, pointing to dissociated levels of activation between the thalamus and the neocortex during ISWS and PS.     

Connexin36 vs. connexin32, "miniature" neuronal gap junctions, and limited electrotonic coupling in rodent suprachiasmatic nucleus

Suprachiasmatic nucleus (SCN) neurons generate circadian rhythms, and these neurons normally exhibit loosely-synchronized action potentials. Although electrotonic coupling has long been proposed to mediate this neuronal synchrony, ultrastructural studies have failed to detect gap junctions between SCN neurons. Nevertheless, it has been proposed that neuronal gap junctions exist in the SCN; that they consist of connexin32 or, alternatively, connexin36; and that connexin36 knockout eliminates neuronal coupling between SCN neurons and disrupts circadian rhythms. We used confocal immunofluorescence microscopy and freeze-fracture replica immunogold labeling to examine the distributions of connexin30, connexin32, connexin36, and connexin43 in rat and mouse SCN and used whole-cell recordings to re-assess electrotonic and tracer coupling. Connexin32-immunofluorescent puncta were essentially absent in SCN but connexin36 was relatively abundant. Fifteen neuronal gap junctions were identified ultrastructurally, all of which contained connexin36 but not connexin32, whereas nearby oligodendrocyte gap junctions contained connexin32. In adult SCN, one neuronal gap junction was >600 connexons, whereas 75% were smaller than 50 connexons, which may be below the limit of detectability by fluorescence microscopy and thin-section electron microscopy. Whole-cell recordings in hypothalamic slices revealed tracer coupling with neurobiotin in <5% of SCN neurons, and paired recordings (>40 pairs) did not reveal obvious electrotonic coupling or synchronized action potentials, consistent with few neurons possessing large gap junctions. However, most neurons had partial spikes or spikelets (often <1 mV), which remained after QX-314 [N-(2,6-dimethylphenylcarbamoylmethyl)triethylammonium bromide] had blocked sodium-mediated action potentials within the recorded neuron, consistent with spikelet transmission via small gap junctions. Thus, a few "miniature" gap junctions on most SCN neurons appear to mediate weak electrotonic coupling between limited numbers of neuron pairs, thus accounting for frequent detection of partial spikes and hypothetically providing the basis for "loose" electrical or metabolic synchronization of electrical activity commonly observed in SCN neuronal populations during circadian rhythms.     

Gene expression in the rat cerebral cortex: comparison of recovery sleep and hypnotic-induced sleep

Most hypnotic medications currently on the market target some aspect of GABAergic neurotransmission. Although all such compounds increase sleep, these drugs differentially affect the activity of the cerebral cortex as measured by the electroencephalogram. Whereas benzodiazepine medications such as triazolam tend to suppress slow wave activity in the cortex, the GABA(B) ligand gamma-hydroxybutyrate greatly enhances slow wave activity and the non-benzodiazepine, zolpidem, which binds to the omega1 site on the GABA(A) receptor/Cl(-) ionophore complex, is intermediate in this regard. Our previous studies have demonstrated that a small number of genes exhibit increased expression in the cerebral cortex of the mouse and rat during recovery sleep after sleep deprivation: egr-3, fra-2, grp78, grp94, ngfi-b, and nr4a3. Using these genes as a panel of biomarkers associated with sleep, we asked whether hypnotic medications induce similar molecular changes in the rat cerebral cortex to those observed when both sleep continuity and slow wave activity are enhanced during recovery sleep. We find that, although each drug increases the expression of a subset of genes in the panel of biomarkers, no drug fully replicates the molecular changes in the cortex associated with recovery sleep. Furthermore, high levels of slow wave activity in the cortex are correlated with increased expression of fra-2 whereas the expression of grp94 is correlated with body temperature. These results demonstrate that sleep-related changes in gene expression may be affected by physiological covariates of sleep and wakefulness rather than by vigilance state per se.     

Normal sleep homeostasis and lack of epilepsy phenotype in GABA A receptor alpha3 subunit-knockout mice

Thalamo-cortical networks generate specific patterns of oscillations during distinct vigilance states and epilepsy, well characterized by electroencephalography (EEG). Oscillations depend on recurrent synaptic loops, which are controlled by GABAergic transmission. In particular, GABA A receptors containing the alpha3 subunit are expressed predominantly in cortical layer VI and thalamic reticular nucleus (nRT) and regulate the activity and firing pattern of neurons in relay nuclei. Therefore, ablation of these receptors by gene targeting might profoundly affect thalamo-cortical oscillations. Here, we investigated the role of alpha3-GABA A receptors in regulating vigilance states and seizure activity by analyzing chronic EEG recordings in alpha3 subunit-knockout (alpha3-KO) mice. The presence of postsynaptic alpha3-GABA A receptors/gephyrin clusters in the nRT and GABA A-mediated synaptic currents in acute thalamic slices was also examined. EEG spectral analysis showed no difference between genotypes during non rapid-eye movement (NREM) sleep or at waking-NREM sleep transitions. EEG power in the spindle frequency range (10-15 Hz) was significantly lower at NREM-REM sleep transitions in mutant compared with wild-type mice. Enhancement of sleep pressure by 6 h sleep deprivation did not reveal any differences in the regulation of EEG activities between genotypes. Finally, the waking EEG showed a slightly larger power in the 11-13-Hz band in alpha3-KO mice. However, neither behavior nor the waking EEG showed alterations suggestive of absence seizures. Furthermore, alpha3-KO mice did not differ in seizure susceptibility in a model of temporal lobe epilepsy. Strikingly, despite the disruption of postsynaptic gephyrin clusters, whole-cell patch clamp recordings revealed intact inhibitory synaptic transmission in the nRT of alpha3-KO mice. These findings show that the lack of alpha3-GABA(A) receptors is extensively compensated for to preserve the integrity of thalamo-cortical function in physiological and pathophysiological situations.     

Schlafregulation

Circadian rhythmicity and sleep homeostasis both contribute to sleep timing and sleep structure in animals and humans. The circadian process and the sleep homeostat interact to consolidate the sleep-wake cycle and, thus, establish wakefulness and sleep. The circadian process generates a sleep-wake propensity rhythm that is timed to oppose homeostatic changes in sleep drive. Disruption of this fined-tuned interaction can lead to performance decrements, daytime sleepiness, and sleep problems, which are often found in shift workers, jet lag, in older people, and patients suffering from delayed or advanced sleep phase syndrome. Recent progress in molecular biology and cell physiology has led to the following conclusions regarding these two processes and their impact on the neurobiology of sleep: The suprachiasmatic nuclei (SCN), located in the anterior hypothalamus, represent the master circadian pacemaker. There is a feedback to the SCN via the neurohormone melatonin. The ventrolateral preoptic area (VLPO) is particularly important for the initiation of sleep. Adenosine triggers the VLPO. An ultradian oscillator located in the mesopontine brainstem region controls the regular cycling between non-REM and REM sleep. The sleep-wake cycle and the NREM-REM sleep cycle induce regularly occurring neuromodulatory changes in forebrain structures.     

A new method for detecting state changes in the EEG: exploratory application to sleep data

A new statistical method is described for detecting state changes in the electroencephalogram (EEG), based on the ongoing relationships between electrode voltages at different scalp locations. An EEG sleep recording from one NREM-REM sleep cycle from a healthy subject was used for exploratory analysis. A dimensionless function defined at discrete times t_i, u(t_i), was calculated by determining the log-likelihood of observing all scalp electrode voltages under the assumption that the data can be modeled by linear combinations of stationary relationships between derivations. The u(t_i), calculated by using independent component analysis, provided a sensitive, but non-specific measure of changes in the global pattern of the EEG. In stage 2, abrupt increases in u(t_i) corresponded to sleep spindles. In stages 3 and 4, low frequency (≈ 0.6 Hz) oscillations occurred in u(t_i) which may correspond to slow oscillations described in cellular recordings and the EEG of sleeping cats. In stage 4 sleep, additional irregular very low frequency (≈ 0.05–0.2 Hz) oscillations were observed in u(t_i) consistent with possible cyclic changes in cerebral blood flow or changes of vigilance and muscle tone. These preliminary results suggest that the new method can detect subtle changes in the overall pattern of the EEG without the necessity of making tenuous assumptions about stationarity.     

Cav2.3 E-/R-type voltage-gated calcium channels modulate sleep in mice

Mammalian sleep is characterized by cycles of REM and non-REM (NREM), i.e. slow-wave sleep (SWS) phases. The major neuroanatomical basis of SWS is the thalamocortical circuitry, which operates in different functional modes to determine the state of vigilance. At high vigilance, the tonic mode predominates; stages of low vigilance and SWS are characterized by rebound burst firing. Electrophysiologically, rebound bursting depends on low-threshold Ca^2?+ spikes and T-type Ca^2?+ channels have been shown to modulate SWS. We recently demonstrated that Ca_v2.3 R-type Ca^2?+ channels are capable of modulating absence seizures, a pathophysiological aberration of the thalamocortical oscillations related to SWS. We thus analyzed sleep architecture in control and Ca_v2.3(?|?) mice using implantable electroencephalography (EEG)/electromyography (EMG) radiotelemetry during spontaneous and urethane-induced sleep. The results demonstrate significantly reduced total sleep time and impairment of SWS generation in Ca_v2.3(?|?) mice, which affects global sleep architecture (i.e. the ratio of REM to NREM). Furthermore, the relative δ power is significantly reduced in Ca_v2.3(?|?) mice during NREM sleep although these mice display longer prior wakefulness, possibly indicating disturbances in sleep homeostasis. This observation is supported by recordings following urethane administration. This is the first study to shed light on the fundamental role of Ca_v2.3 channels in rodent sleep physiology.     

The two‐process model of sleep regulation: a reappraisal

In the last three decades the two‐process model of sleep regulation has served as a major conceptual framework in sleep research. It has been applied widely in studies on fatigue and performance and to dissect individual differences in sleep regulation. The model posits that a homeostatic process (Process S) interacts with a process controlled by the circadian pacemaker (Process C), with time‐courses derived from physiological and behavioural variables. The model simulates successfully the timing and intensity of sleep in diverse experimental protocols. Electrophysiological recordings from the suprachiasmatic nuclei (SCN) suggest that S and C interact continuously. Oscillators outside the SCN that are linked to energy metabolism are evident in SCN‐lesioned arrhythmic animals subjected to restricted feeding or methamphetamine administration, as well as in human subjects during internal desynchronization. In intact animals these peripheral oscillators may dissociate from the central pacemaker rhythm. A sleep/fast and wake/feed phase segregate antagonistic anabolic and catabolic metabolic processes in peripheral tissues. A deficiency of Process S was proposed to account for both depressive sleep disturbances and the antidepressant effect of sleep deprivation. The model supported the development of novel non‐pharmacological treatment paradigms in psychiatry, based on manipulating circadian phase, sleep and light exposure. In conclusion, the model remains conceptually useful for promoting the integration of sleep and circadian rhythm research. Sleep appears to have not only a short‐term, use‐dependent function; it also serves to enforce rest and fasting, thereby supporting the optimization of metabolic processes at the appropriate phase of the 24‐h cycle.     

Polygraphic and behavioral study of sleep in geese: existence of nuchal atonia during paradoxical sleep

In adult geese, chronic polygraphic recordings of EEG, EOG, EMG, ECG and respiratory rate completed with behavioral observations allowed the characterization of four states of vigilance: wakefulness (W), drowsiness (D), slow wave sleep (SWS) and paradoxical sleep (PS). The EEG, EOG, EMG general patterns observed during W, D, SWS and PS episodes with nuchal isotonia or hypotonia were similar to those reported in other birds. The characteristic brevity of avian PS was confirmed since this sleep state occupied only 2.8% of the nycthemere in geese. For the first time in an adult bird it was shown that numerous PS episodes were accompanied, as in mammals, by a total disappearance of nuchal EMG activity. These observations made in a bird species with a stable head support when sleeping, suggest that, as in mammals, inhibitory mechanisms leading to a PS related nuchal atonia do exist and that head falling is not the cause of PS episodes brevity in birds.     

Automatic classification of the cats vigilance state.

A special purpose hardware system has been interfaced with a minicomputer for determining the vigilance state in the cat and for quantifying delta and sigma spindle activity in the sleep EEG. The computer agreement with manual scoring is 93.8% when the state is classified as awake. non-REM, or REM sleep. The system identifies six distinct vigilance states: REM sleep, slow-wave sleep 1 or 2 (distinguished by differences in EMG level), light sleep, resting and movement. Five day averages of vigilance state and EEG activity are presented for three animals, and the effects of 5 HTP and L-dopa on delta activity and sleep spindles have been quantified for one cat in order to demonstrate the system utility.     

Diurnal changes in electrocorticogram sleep slow‐wave activity during development in rats

According to the homeostatic regulation of sleep, sleep pressure accumulates during wakefulness, further increases during sleep deprivation and dissipates during subsequent sleep. Sleep pressure is electrophysiologically reflected by electroencephalogram slow‐wave activity during non‐rapid eye movement sleep, and is thought to be stable across time. During childhood and adolescence the brain undergoes massive reorganization processes. Slow‐wave activity during these developmental periods has been shown in humans to follow an inverted U‐shaped trajectory, which recently was replicated in rats. The goal of this study was to investigate in rats the diurnal changes of slow‐wave activity during the inverted U‐shaped developmental trajectory of slow‐wave activity. To do so, we performed longitudinal electrocorticogram recordings, and compared the level of slow‐wave activity at the beginning with the slow‐wave activity level at the end of 24‐h baselines in two sets of Sprague–Dawley rats. In younger animals (n = 17) we investigated specific postnatal days when overall slow‐wave activity increases (postnatal day 26), peaks (postnatal day 28) and decreases (>postnatal day 28). The same analysis was performed in older animals (postnatal day 48, n = 6). Our results show a gain of slow‐wave activity across 24 h on postnatal day 26, followed by no net changes on postnatal day 28, which was then followed by a loss of slow‐wave activity during subsequent days (>postnatal day 28). Older animals did not show any net changes in slow‐wave activity across 24 h. These results cannot be explained by differences in vigilance states. Thus, slow‐wave activity during this developmental period may not only reflect the trajectory of sleep pressure but may additionally reflect maturational processes.     

Sleep characteristics in the turkey Meleagris gallopavo

Electrophysiological and behavioral characteristics of the states of vigilance were analyzed in chronically implanted specimens of the turkey Meleagris gallopavo (M. gallopavo). Five different states of vigilance were observed throughout the nyctohemeral period: active wakefulness (AW), quiet wakefulness (QW), drowsiness (D), slow wave sleep (SWS) and rapid eye movement (REM) sleep.These states exhibit characteristics similar to those described in other bird species. Sleep periods displayed a polyphasic distribution; however, they showed the tendency to concentrate between 2100 and 0900 h in spite of the fact that the recordings were carried out under constant illumination. Sleep period occupied 45.71% of the nyctohemeral cycle, 43.33% corresponded to SWS, while 2.38% to REM sleep. The average duration of the REM sleep phase was very short, lasting 7.7+/-0.55 s (mean+/-S.D.). In contrast, its frequency was very high with an average recurrence of 268+/-63 phases throughout the nyctohemeral cycle. The short duration of REM sleep phase presented by the turkey as by other bird species studied up to now may be dependent upon genetic factors shared by this group of vertebrates.     

Clock controls circadian period in isolated suprachiasmatic nucleus neurons

The suprachiasmatic nucleus (SCN) is the master circadian pacemaker in mammals, and one molecular regulator of circadian rhythms is the Clock gene. Here we studied the discharge patterns of SCN neurons isolated from Clock mutant mice. Long-term, multielectrode recordings showed that heterozygous Clock mutant neurons have lengthened periods and that homozygous Clock neurons are arrhythmic, paralleling the effects on locomotor activity in the animal. In addition, cells in dispersals expressed a wider range of periods and phase relationships than cells in explants. These results suggest that the Clock gene is required for circadian rhythmicity in individual SCN cells and that a mechanism within the SCN synchronizes neurons and restricts the range of expressed circadian periods.     

Spontaneous activity of the perirhinal cortex in behaving cats.

The perirhinal cortex lies at the interface between the neocortex and allocortex. Whether the perirhinal cortex expresses spontaneous electroencephalographic rhythms that are characteristic of the allocortex and/or of the neocortex is unknown. Thus, the present investigation was undertaken to characterize the activity of the perirhinal cortex with respect to various electroencephalographic rhythms that are displayed by neocortical areas or the entorhino-hippocampal system during different behavioral states of vigilance in chronically-implanted cats. Although perirhinal and neocortical electroencephalograms underwent similar state-dependent changes in amplitude, the ubiquitous neocortical sleep spindles were absent from the perirhinal cortex. In addition, while the slow sleep oscillation (0.5-1 Hz), which is pervasive in the neocortex, was present in the perirhinal cortex, its temporal relation to the neocortical oscillation was highly variable. In contrast, a high degree of correlation was found between perirhinal and entorhinal electroencephalographic activities in all behavioral states. In particular, during waking and paradoxical sleep, multiple simultaneously recorded entorhinal and perirhinal sites displayed an oscillation in the theta range which was highly correlated. To rule out the possibility that the perirhinal theta oscillation reflected volume conduction from neighboring structures, single-unit recordings were performed. Spike-triggered averages and peri-event histograms revealed that perirhinal cells displayed a statistically significant theta-related modulation of their spontaneous activity, albeit weaker than that observed in the entorhinal cortex. Thus, from the standpoint of spontaneous electroencephalographic rhythms, the perirhinal cortex is more closely related to the entorhino-hippocampal system than to the neocortex.