Sleep wake profile and EEG spectral power in young or old senescence accelerated mice

Changes occurring with age in cortical EEG and sleep-wake states architecture were examined in senescence accelerated prone (SAMP8) or senescence resistant (SAMR1) mice (age: 2 and 12 months) under baseline conditions or after a 4 h sleep deprivation (SD). In baseline conditions, an increase in slow wave sleep (SWS) amount (21-24%) occurs at the expense of the wakefulness (W) in old SAMP8 and SAMR1 mice versus young animals. In these conditions, SWS latency is reduced (67-72%). Moreover, in SAMP8 and SAMR1 mice, aging deteriorates paradoxical sleep (PS) architecture with more pronounced changes in SAMP8 (amount: -63%; episode duration: -44%; latency: +286%; circadian component loss; and EEG theta (theta) peak frequency (TPF): -1 Hz). During the 4 h recovery subsequent to a 4 h sleep deprivation, old SAMP8 mice exhibit an enhanced sensitivity resulting in SWS (+62%) and PS (+120%) rebounds, a characteristic of this inbred strain. Results obtained are discussed in line with the age-related learning and memory impairments existing in SAMP8 animals. In particular, the reduced cognitive performances described in old SAMP8 might be linked to the TPF deterioration during PS.     

Nitric oxide and sleep in the rat: a puzzling relationship.

To date, only a few studies indicate that nitric oxide may play a role in the regulation of the sleep-wake cycle. However, data reported are controversial and the part played by nitric oxide in sleep-wake cycle regulation still remains uncertain. In the present report, we studied the effects on sleep amounts of two different nitric oxide synthase inhibitors: N-nitro-L-arginine methyl ester, a non-selective nitric oxide synthase inhibitor, and 7-nitro-indazole, a specific inhibitor of neuronal nitric oxide synthase. The above compounds were administered via two routes, i.e. intraperitoneally or locally in the dorsal raphe nucleus, a structure involved in sleep regulation. In order to evaluate their efficiency to inhibit nitric oxide synthesis in the rat brain, they were first administered intraperitoneally to a group of animals, and the cortical release of nitric oxide was determined by means of voltammetric measurements. N-Nitro-L-arginine methyl ester (100 mg/kg, i.p.) did not affect the cortical release of nitric oxide, whereas it increased both slow-wave sleep and paradoxical sleep durations. On the contrary, 7-nitro-indazole (40 mg/kg, i.p.) significantly decreased the cortical release of nitric oxide (-25%) and paradoxical sleep duration. Furthermore, following microinjection of either N-nitro-L-arginine methyl ester or 7-nitro-indazole at 100 ng/0.20 microl into the nitric oxidergic cell area of the dorsal raphe nucleus, decreases in paradoxical sleep duration were obtained (-32.8% and -25.3%, respectively). The results obtained support the existence of a duality in the sleep regulation modalities exerted by nitric oxide, i.e. a peripheral inhibiting influence and a central facilitating role for the nitric oxide-serotoninergic neurons of the dorsal raphe nucleus.     

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.     

Potential role of inducible nitric oxide synthase in the sleep-wake states occurrence in old rats

Extensive evidences now suggest that an association between inducible nitric oxide synthase and oxidative stress takes place during aging. Since the part played by inducible nitric oxide synthase in the sleep impairments associated with aging still remains unexplored, we compared its involvement in old rats (20-24 months) versus adult ones (3-5 months) using polygraphic, biochemical, voltammetric and immunohistochemical techniques. The experiments were conducted either in basal condition or after a systemic injection of selected inducible nitric oxide synthase inhibitors. We found that 2-amino-5,6-dihydro-6-methyl-4H-1,3-thiazine (10 mg/kg, i.p.) or aminoguanidine (400 mg/kg, i.p.) was capable to suppress rapid-eye-movement sleep and induce a delayed enhancement in slow-wave sleep in old rats. These effects did not occur in adult animals. Within the frontal cortex, the laterodorsal tegmentum and dorsal raphe nuclei, the basal inducible nitric oxide synthase activity was 85-200% higher in old rats than in adult ones. In contrast, the neuronal nitric oxide synthase activity did not vary in both groups. 2-Amino-5,6-dihydro-6-methyl-4H-1,3-thiazine administration significantly reduced inducible nitric oxide synthase activity (70-80% according to the brain areas) independently of age, but significantly decreased the cortical nitric oxide release in old rats. Finally, in frontal cortex and dorsal raphe immunohistochemical analysis showed inducible nitric oxide synthase-positive cells again only in old animals. These data support the idea that nitric oxide produced by inducible nitric oxide synthase plays a role in the triggering and maintenance of rapid-eye-movement sleep during aging.     

An EEG averaging technique for automated sleep-wake stage identification in the rat

An automated on-line sleep-wake classification system based on an averaging technique of the running EEG is described. It operates for three rats simultaneously and is able to discriminate every 5 sec between wakefulness, light slow-wave sleep, deep slow-wave sleep, and paradoxical sleep. The hippocampal EEG and nuchal EMG are used as input parameters. The EEG is bandpass filtered after which a microcomputer samples and averages the filtered EEG and constructs spectrograms. The variability, the theta-delta ratio and the amplitude of the delta waves are obtained from these spectrograms. Together with the amplitude index of the EMG, these three EEG indices are subjected to decision rules for the identification of sleep-wake states. A first evaluation study showed 93% agreement between visual inspection and computer classification. In a second evaluation study 24-hr recordings were made. Clear circadian patterns emerged especially during the light period: deep slow-wave sleep was enhanced during the initial hours, while paradoxical sleep tended to increase over the latter hours. The outcome of these studies is compared with the results obtained with other automated sleep identification procedures.     

Circadian regulation of arousal: role of the noradrenergic locus coeruleus system and light exposure

STUDY OBJECTIVES: Noradrenergic locus coeruleus (LC) neurons regulate arousal. Previous studies have shown that noradrenergic LC neurons exhibit a circadian rhythm in impulse activity, which peaks during the active period. This is mediated by an indirect circuit projection from the suprachiasmatic nucleus (SCN) to the LC. Here we sought to evaluate the hypothesis that the LC regulates the circadian properties of the sleep-wake cycle. DESIGN: Sprague-Dawley rats maintained on a light-dark (LD) schedule or in constant darkness (DD) for 3 to 4 weeks were treated with DSP-4, a neurotoxic agent specific for noradrenergic-LC projections. Vigilance states were analyzed before and 3 weeks after LC lesion. The DSP-4 lesion was verified by immunohistochemistry of noradrenergic fibers in the frontal cortex. SETTING: University of Pennsylvania. PATIENTS OR PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: DSP-4 decreased the amplitude of the sleep-wake rhythm in LD animals by significantly decreasing wakefulness and increasing sleep during the active period. However, DSP-4 had no effect on the sleep-wake cycle of DD animals. Moreover, DD itself decreased the amplitude of the sleep-wake cycle similar to that of the neurotoxic lesion of the noradrenergic system in LD animals. Analysis of noradrenergic fiber staining in the frontal cortex revealed that this effect was associated with fewer fibers or boutons in nonlesioned DD rats than in nonlesioned LD animals. CONCLUSIONS: Noradrenergic LC neurons provide a circadian regulation of the sleep-wake cycle, and the maintenance of LC function depends on light exposure. Light deprivation induces a loss of noradrenergic fibers, which in turn decreases the amplitude of the sleep-wake rhythm.     

Sleep deprivation in the rat: IX. Recovery

Eight rats were subjected to total sleep deprivation, paradoxical sleep deprivation, or high amplitude sleep deprivation until they showed major deprivation-induced changes. Then they were allowed to sleep ad lib. Three rats that had shown the largest temperature declines died within two to six recovery days. During the first 15 days of ad lib sleep, surviving rats showed complete or almost complete reversal of the following deprivation-induced changes: debilitated appearance, lesions on the paws and tail, high energy expenditure, large decreases in peritoneal temperature, high plasma epinephrine and norepinephrine levels, and low thyroxine levels. The most prominent features of recovery sleep in all rats were immediate and large rebounds of paradoxical sleep to far above baseline levels, followed by lesser temporally extended rebounds. Rebounds of high amplitude non-rapid eye movement (NREM) sleep occurred only in some rats and were smaller and less immediate.     

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.     

Effects of sleep deprivation on sleepiness, sleep intensity, and subsequent sleep in the rat

The effects of 24 hr of sleep deprivation on cortical EEG and ventral hippocampus EEG recordings, ventral hippocampus spike rates, sleep stages percentages, and bout length measures were studied in rats. Two groups, differing only in the rate and distance they were forced to walk during deprivation by the water wheel method, were recorded continuously (23 hr per day) for one baseline, one deprivation, and two recovery days. During deprivation, microsleeps, increased hippocampal spike rates, and increased amplitude of the EEG recordings all suggested the intrusion of sleep processes. Nonetheless, there was no evidence to support the idea that these animals were not substantially deprived of sleep. No important differences were found in the recovery data of the two groups, even though one group walked three times as far as the other during deprivation. This supports the idea that, in conjunction with large amounts of sleep deprivation, changes in exercise and energy depletion may have little effect on sleep measures. During recovery, increased hippocampal spike rates and bout lengths, as well as increases in EEG amplitude, were interpreted in terms of increased sleep "intensity." High amplitude NREM sleep rebounded first, followed by rebounds in both paradoxical sleep and low amplitude NREM sleep. This pattern was compared to patterns previously reported for humans, cats, and rats. Finally, the tendency for some measures to fall below their baseline levels after an initial rebound was discussed in terms of "sleep inhibition" and servomechanism theory.     

The pendulum technique for paradoxical sleep deprivation in rats

A new technique for paradoxical sleep (PS) deprivation in rats is presented. Animals are prevented from entering into PS by allowing them to sleep for only brief periods of time. This is accomplished by an apparatus which moves the animals' cages backwards and forwards like a pendulum. At the extremes of the motion postural imbalance is produced in the animals forcing them to walk downwards to the other side of their cages. A minimal amount of PS and a moderate amount of slow wave sleep (SWS) were detected during a deprivation period of 72 hrs. Following the deprivation treatment the recovery of sleep was monitored for 3 hrs; at the beginning of the light period for one group and at the beginning of the dark period for a second group. The sleep-waking patterns of two baseline groups were established at the time when the recovery sleep was examined in the deprivation groups. The deprivation treatment resulted in a significant increase in the amount of PS and a significant decrease in the amount of SWS. The extent of PS increase was similar in both deprivation groups, in spite of a large difference in the amount of SWS. The decrease of SWS mainly occurred during recovery sleep in the light. It was observed that sleep in the dark differs from sleep in the light in behavioural aspects.     

Paradoxical (REM) sleep deprivation in mice using the small‐platforms‐over‐water method: polysomnographic analyses and melanin‐concentrating hormone and hypocretin/orexin neuronal activation before,during and after deprivation

Studying paradoxical sleep homeostasis requires the specific and efficient deprivation of paradoxical sleep and the evaluation of the subsequent recovery period. With this aim, the small‐platforms‐over‐water technique has been used extensively in rats, but only rare studies were conducted in mice, with no sleep data reported during deprivation. Mice are used increasingly with the emergence of transgenic mice and technologies such as optogenetics, raising the need for a reliable method to manipulate paradoxical sleep. To fulfil this need, we refined this deprivation method and analysed vigilance states thoroughly during the entire protocol. We also studied activation of hypocretin/orexin and melanin‐concentrating hormone neurones using Fos immunohistochemistry to verify whether mechanisms regulating paradoxical sleep in mice are similar to those in rats. We showed that 48 h of deprivation was highly efficient, with a residual amount of paradoxical sleep of only 2.2%. Slow wave sleep and wake quantities were similar to baseline, except during the first 4 h of deprivation, where slow wave sleep was strongly reduced. After deprivation, we observed a 124% increase in paradoxical sleep quantities during the first hour of rebound. In addition, 34% of hypocretin/orexin neurones were activated during deprivation, whereas melanin‐concentrated hormone neurones were activated only during paradoxical sleep rebound. Corticosterone level showed a twofold increase after deprivation and returned to baseline level after 4 h of recovery. In summary, a fairly selective deprivation and a significant rebound of paradoxical sleep can be obtained in mice using the small‐platforms‐over‐water method. As in rats, rebound is accompanied by a selective activation of melanin‐concentrating hormone neurones.     

Effects of aging and housing in an enriched environment on sleep-wake patterns in rats

The effects of aging and housing in an enriched environment were assessed in young adult (4-7 months) and old (27-31 months) male Brown Norway rats by conducting 24-h sleep-wake recordings. Comparison of recordings made in rats of different ages, housed in a standard laboratory environment, revealed a reduction of the time spent in slow wave and desynchronized sleep during the light period in the old rats. Furthermore in the old rats, sleep was more fragmented and the amplitude of the circadian sleep-wake rhythm was reduced. In both age groups, housing in an enriched environment resulted in an increase of the time spent in slow wave and desynchronized sleep during the light period. Old "enriched" rats showed an additional alleviation of the senescence-related shortening of sleep cycles and desynchronized sleep epochs. The reduction of the circadian sleep-wake amplitude observed in old age was, however, not affected by the differential housing period. It is concluded that the similarity of the changes in sleep pattern in young and old rats after increased environmental complexity may reflect a preserved capacity of the senescent nervous system to adapt to environmental changes.     

Effects of pinealectomy on baseline sleep and response to sleep deprivation

STUDY OBJECTIVES: We have previously reported that older (24 mo.) Fischer rats manifest a diminished post-sleep deprivation increase in NREM and REM sleep. In order to examine whether this decline reflects an age-related change in pineal function, we are now reporting on baseline and recovery sleep parameters in pinealectomized 3-, 12-, and 24-month old rats following 24 hours of sleep deprivation using the disk-over-water method. DESIGN: Three independent age groups; within each group there were sequential measures of sleep under baseline conditions and during recovery from sleep deprivation. SETTING: The Sleep Research Laboratory at the University of Chicago PARTICIPANTS: 56 male Fisher (F344) rats INTERVENTIONS: 24 hours of total sleep deprivation using the disk-over-water method MEASUREMENTS: Sleep staging of EEG and EMG, and power spectral analysis of the EEG RESULTS: Pinealectomized (pinex) rats did not differ from sham-operated (sham) rats in total sleep, REM sleep, super-modal high-amplitude NREM sleep (HS2), a measure of NREM EEG delta power, or circadian rhythm amplitude. In the pinex rats, there was a modest (2.5%) age-independent increase in NREM sleep (p<0.02). The pinex rats of all ages failed to manifest the increase in NREM sleep during recovery seen in the sham-operated animals (p<0.04). CONCLUSIONS: We found no evidence that altered pineal function is responsible for age-related changes in baseline sleep in the rat. These data also suggest that, independent of age, normal pineal function may be relevant to the ability to generate increased NREM sleep in response to prior sleep deprivation.     

Age-dependent changes in recovery sleep after 48 hours of sleep deprivation in rats

To characterize possible changes in homeostatic regulation of sleep with aging, we have examined sleep stages during recovery sleep after 48 h of sleep deprivation in young (3 months), middle aged (12 months), and old (24 months) rats. It was found that young and middle aged, in contrast to old rats, had large (21-24%) increases in total sleep time during recovery sleep; the old rats experienced a quantitatively small (8%) but significant rise in total sleep. NREM sleep increased significantly during the recovery period in young and middle aged, but not older rats. High voltage NREM sleep (HS2) declined by 30% during recovery in the young animals, but remained unchanged compared to baseline in the middle aged and old animals. The young and middle aged rats had increases in REM sleep during recovery compared to their baseline by 96% and 93%, respectively, which was significantly greater than a 65% increase during recovery in the old rats. Increases in total sleep and REM sleep during recovery were largely confined to the first 6 h in young and middle aged rats, but maxima for the old rats occurred in the second 6 h.     

Effects of lesions of the suprachiasmatic nuclei on paradoxical sleep and slow wave sleep circadian rhythms in the rat

Continuous polygraphic recordings were taken from four rats before and after total lesions of the suprachiasmatic nuclei. This lesion did not change the total amount of paradoxical sleep nor slow wave sleep over a 24-h period, but reduced the amplitude of their circadian variations by 50%. Control lesions rostral and caudal to the suprachiasmatic nuclei were without effect on these sleep stages.     

Sleep and memory relationships in intact old and amnestic young rats

Age-related changes in sleep are observed in many species, including rats and humans. Old rats often exhibit less total and paradoxical sleep, shorter sleep bouts and more random sleep-wake periods across 24 hours, than young rats. This paper evaluates recent evidence that deterioration of selected sleep parameters, usually involving levels of paradoxical sleep or durations of sleep bouts, may be related to deterioration of memory in old rats. Similar findings are reviewed with respect to young animals with different forms of experimentally-induced amnesia. Furthermore, a drug that enhances memory in rats and old humans, glucose, also enhances paradoxical sleep in old rats. These data suggest the utility of sleep measures as neurobiological markers of memory dysfunction in old rats.     

Comparison of sustained attention assessed by auditory and visual psychomotor vigilance tasks prior to and during sleep deprivation

To date, no detailed examination of the pattern of change in reaction time performance for different sensory modalities has been conducted across the circadian cycle during sleep deprivation. Therefore, we compared sustained auditory and visual attention performance during 40 h of sleep deprivation assessing multiple metrics of auditory and visual psychomotor vigilance tasks (PVT). Forty healthy participants (14 women) aged 30.8 ± 8.6 years were studied. Subjects were scheduled for an ∼8 h sleep schedule at home prior to three–six laboratory baseline days with an 8 h sleep schedule followed by 40 h sleep deprivation. Visual and auditory PVTs were 10 min in duration, and were administered every 2 h during sleep deprivation. Data were analysed with mixed‐model anova . Sleep deprivation and circadian phase increased response time, lapses, anticipations, standard deviation of response times and time on task decrements for visual and auditory PVTs. In general, auditory vigilance was faster and less variable than visual vigilance, with larger differences between auditory and visual PVT during sleep deprivation versus baseline. Failures to respond to stimuli within 10 s were four times more likely to occur to visual versus auditory stimuli. Our findings highlight that lapses during sleep deprivation are more than just long responses due to eye closure or visual distraction. Furthermore, our findings imply that the general pattern of change in attention during sleep deprivation (e.g. circadian variation, response slowing, lapsing and anticipations, time on task decrements and state instability) is similar among sensory–motor behavioral response modalities.     

Larger phase angle between sleep propensity and melatonin rhythms in sighted humans with non-24-hour sleep-wake syndrome

STUDY OBJECTIVES: This study was aimed to clarify phase angle between sleep propensity and the circadian pacemaker in patients with non-24-hour sleep-wake syndrome (Non-24). DESIGN AND SETTING: A case-control study was underaken. PARTICIPANTS: Sighted patient with Non-24 (4 males and 1 female, aged 16 to 39 y), and sex- and age-matched healthy controls (12 males and 3 females, aged 19 to 35 y) participated the study. MEASUREMENT AND INTERVENTION: Following an actigraphic assessment of the sleep-wake cycle in their homes, the participants entered an ultra-short sleep-wake schedule together with simultaneous measurement of dim light melatonin rhythm after 24-hour sleep deprivation. RESULTS: The period of the sleep-wake cycle observed at home was longer in the Non-24 patients (25.12 hours) than in the controls (24.02 hours, p<0.0001). The interval from sleep propensity (SP) onset to the melatonin midpoint (MLmid) was significantly shorter in the Non-24 patients than in the controls. The interval from the MLmid to the SP offset was significantly longer in the Non-24 patients than in the controls. CONCLUSIONS: It was postulated that Non-24 sufferers' delayed SP onset relative to the circadian pacemaker may accelerate the light-induced phase-delay, leading to sleep-wake cycle that is longer than 24 hours.     

Sleep deprivation in the rat: XVI. Effects in a light-dark cycle.

To avoid a possible confound between the effects of sleep loss and disturbed circadian rhythms in previous studies of total sleep deprivation (TSD) by the disk-over-water method, TSD rats and their yoked control (TSC) rats had been maintained in constant light both before and during the experiment. With circadian rhythms of both groups flattened by constant light, group differences in outcome measures could be attributed to sleep loss. However, the constant light control entailed the possibility that the sleep loss effects might obtain only in constant light. To evaluate this possibility, three TSD-TSC rat pairs maintained on a 12 hour light: 12 hour dark (LD) schedule were studied. TSC rats showed only minor changes during the deprivation period. As in previous studies, TSD rats showed increased food intake; decreased weight; increased energy expenditure; debilitated appearance; lesions on the tail and paws; an initial increase followed by a large decrease in body temperature; impending death; and recovery sleep, which featured large, selective, sustained rebounds of paradoxical sleep and a reversal of all observed TSD-induced changes. Thus, TSD produced the same changes during an LD schedule as during constant light. The amplitude of the diurnal body temperature rhythm declined over the course of TSD and then almost completely recovered during the first day of recovery sleep. The decline was interpreted as the result of deprivation-induced thermoregulatory changes.