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This study aims to explore the polysomnographically measured sleep and leg movement differences between idiopathic restless legs syndrome (RLS) patients and healthy controls.An electronic literature search was conducted in EMBASE, MEDLINE, all EBM databases, CINAHL, and PsycINFO. Only observational case-control studies were included in the meta-analysis. The differences in 13 sleep parameters and 23 leg movement parameters between RLS patients and healthy controls were explored.Thirty-eight studies were identified for systematic review, 31 of which were used for meta-analysis. Meta-analyses revealed significant reductions in total sleep time, sleep efficiency, stage N2 and rapid eye movement (REM) sleep percentages, and increases in wake time after sleep onset, stage shifts per hour, stage N1 percentage, REM latency, arousal index, and apnea-hypopnea index. Some leg movement parameters, such as periodic limb movement during sleep (PLMS) index, PLMS sequence duration, number of PLMS sequence, and periodicity index, were higher in RLS patients compared with healthy controls. Further, our meta-analysis revealed a higher PLMS index during non-REM sleep compared with that during REM sleep.RLS patients manifest a lightening of sleep, increased sleep fragmentation, and greater sleep-related breathing disruption and limb movements during sleep relative to healthy normal individuals. The distributions of PLMS during a night's sleep may provide more information to clarify the specific characteristics of leg movements in RLS. PLMS in RLS are concentrated in non-REM sleep. The periodicity index may be a more sensitive and specific marker of RLS than the PLMS index.Zhang H, Zhang Y, Ren R, et al. Polysomnographic features of idiopathic restless legs syndrome: a systematic review and meta-analysis of 13 sleep parameters and 23 leg movement parameters.

Previous studies on the association of insomnia with body mass index (BMI) have been controversial. Physiological hyperarousal, the key pathological mechanism of insomnia, may be an important reason for different findings. We explored whether insomnia with physiological hyperarousal measured by the multiple sleep latency test (MSLT) is associated with body-weight differences. A total of 185 normal sleepers and 440 insomniacs were included in this study. Insomnia was defined by standard diagnostic criteria with symptoms lasting ≥6 months. All subjects underwent one night of laboratory polysomnography followed by a standard MSLT. We used the median MSLT value (i.e., ≥14 min) to define physiological hyperarousal. BMI was based on measured height (cm) and weight (kg) during the subjects’ sleep laboratory visit. BMI > 25 kg/m2 was defined as overweight, while BMI 2 was defined as underweight. After controlling for confounders, the odds of lower weight rather than overweight were significantly increased among insomnia patients with increased MSLT: insomnia with MSLT 14–17 min and MSLT > 17 min increased the odds of lower weight by approximately 89% (OR = 1.89, 95% CI 1.00–4.85) and 273% (OR = 3.73, 95% CI 1.51–9.22) compared with normal sleepers, respectively. In contrast, insomnia in patients with MSLT 11–14 min and 8–11 min was not different from normal sleepers in terms of body weight. Insomnia associated with physiological hyperarousal, the most severe phenotype of chronic insomnia, is associated with higher odds of lower weight and underweight compared with normal sleepers. This is a novel finding consistent with previous physiologic data and has significant clinical implications.

Pharmacological and optogenetic studies have demonstrated that the basolateral amygdala (BLA) plays a pivotal role in regulating fear-conditioned changes in sleep, in particular, rapid eye movement sleep (REM). However, the linkage between BLA and REM regulation has been minimally examined. In this study, we optogenetically activated or inhibited BLA selectively during spontaneous REM, and determined the effects on REM amounts and on hippocampus regulated EEG-theta (θ) activity. Excitatory (CaMKIIα-hChR2 (E123A)-eYFP-WPRE) or inhibitory (CaMKIIα-eNpHR3.0-eYFP-WPRE) optogenetic constructs were stereotaxically delivered targeting glutamatergic cells in BLA (BLAGlu) of mice. Viral constructs without opsin (CaMKIIα-eYFP-WPRE) were used as controls. All mice were implanted with telemetry transmitters for monitoring electroencephalography (EEG), activity, and body temperature, and with optic cannulas for light delivery to the BLA. BLAGlu were optogenetically activated by blue light (473 nm), or inhibited by green light (532 nm), in 10 s epochs during REM, or non-REM (NREM), in undisturbed mice. Sleep amounts and EEG activity were analyzed. Projections from BLAGlu to neurons in hippocampus were immunohistochemically (IHC) examined. Brief optogenetic activation of BLAGlu during REM immediately reduced EEG theta activity (5–8 Hz, REM-θ), without affecting overall amount and propensity of sleep, while optogenetic inhibition increased REM-θ. Stimulation during NREM had no effect on EEG spectra or sleep. IHC results showed that glutamatergic and GABAergic cells in CA3 of the hippocampus received inputs from BLAGlu projection neurons. Activation of BLAGlu reduced, and inhibition increased, REM-θ without otherwise altering sleep. Optogenetic stimulation of BLAGlu may be useful for systematically manipulating sleep-related amygdalo-hippocampal interactions.

This study aims to explore the comparative efficacy and acceptability of psychotherapies, pharmacotherapies, and their combinations for insomnia. MEDLINE, EMBASE, Cochrane Central Register of Controlled Trials, PsycINFO, CINAHL, and ClinicalTrials.gov were searched for randomized clinical trials (RCTs) examining the comparative efficacy in which a psychotherapy and a pharmacotherapy for insomnia were directly compared with each other, or in which a combination of psychotherapy and pharmacotherapy was compared with either alone. The analysis included data from 23 RCTs. In 18 of 23 studies, cognitive behavior therapy for insomnia (CBT-I) was the psychotherapy. At post-treatment, CBT-I showed higher subjective sleep efficiency (SE), and lower subjective wake time after sleep onset (WASO) and insomnia severity index (ISI) score. Compared with CBT-I plus pharmacotherapy, pharmacotherapy showed lower subjective SE, and higher subjective sleep latency (SL), PSG measured SL, subjective WASO, and ISI score. Overall, the findings derived from post-treatment data suggested that CBT-I is more beneficial in treating insomnia compared with pharmacotherapy. CBT-I combined with pharmacotherapy is beneficial in improving some sleep parameters (i.e., subjective SE, SL, WASO, and PSG measured SL) compared with pharmacotherapy alone. Daily clinical decisions should consider these findings on the relative efficacy of the principal approaches to insomnia treatment.

The central nucleus of the amygdala (CNA) projects to brainstem regions that generate and regulate rapid eye movement sleep (REM). We used optogenetics to assess the influence of CNA inputs into reticularis pontis oralis (RPO), pedunculopontine tegmentum (PPT) and nucleus subcoeruleus (SubC) on dark period sleep. We compared these results to effects of microinjections into CNA of the GABA(A) agonist, muscimol (inhibition of cell bodies) and tetrodotoxin (TTX, inhibition of cell bodies and fibers of passage). For optogenetics, male Wistar rats received excitatory (AAV5-EF1a-DIO -hChR2(H134R)-EYFP) or inhibitory (AAV-EF1a-DIO-eNpHR3.0-EYFP; DIO-eNpHR3.0) opsins into CNA and AAV5-EF1a-mCherry-IRES-WGA-Cre into RPO, PPT, or SubC. This enabled only CNA neurons synaptically connected to each region to express opsin. Optic cannulae for light delivery into CNA and electrodes for determining sleep were implanted. Sleep was recorded with and without blue or amber light stimulation of CNA. Separate rats received MUS or TTX into CNA prior to recording sleep. Optogenetic activation of CNA neurons projecting to RPO enhanced REM and did not alter non-REM (NREM) whereas activation of CNA neurons projecting to PPT or SubC did not significantly affect sleep. Inhibition of CNA neurons projecting to any region did not significantly alter sleep. TTX inactivation of CNA decreased REM and increased NREM whereas muscimol inactivation did not significantly alter sleep. Thus, the amygdala can regulate decreases and increases in REM, and RPO is important for CNA promotion of REM. Fibers passing through CNA, likely from the basolateral nucleus of the amygdala, also play a role in regulating sleep.

Polysomnographic studies have been conducted to explore nighttime sleep features in narcolepsy, but their relationship to narcolepsy is still imperfectly understood. We conducted a systematic review of the literature exploring polysomnographic differences between narcolepsy patients and healthy controls (HCs) in EMBASE, MEDLINE, All EBM databases, CINAHL, and PsycINFO. 108 studies were identified for this review, 105 of which were used for meta-analysis. Meta-analyses revealed significant reductions in sleep latency, sleep efficiency, slow wave sleep percentage, rapid eye movement sleep (REM) latency, cyclic alternating pattern rate, and increases in total sleep time, wake time after sleep onset (WASO), awakening numbers (AWN) per hour, stage shift (SS) per hour, N1 percentage, apnea hypopnea index, and periodic limb movement index in narcolepsy patients compared with HCs. Furthermore, narcolepsy type 1 patients showed more disturbed nighttime sleep compared with narcolepsy type 2 patients. Children and adolescent narcolepsy patients show increased WASO, AWN, and SS compared with adult patients. Macro- and micro-structurally, our study suggests that narcolepsy patients have poor nighttime sleep. Sex, age, body mass index, disease duration, disease type, medication status, and adaptation night are demographic, clinical and methodological factors that contribute to heterogeneity between studies.

Fear conditioning associated with inescapable shock training (ST) and fearful context re-exposure (CR) alone can produce significant behavioral fear, a stress response and alterations in subsequent REM sleep. These alterations may vary among animals and are mediated by the basolateral nucleus of the amygdala (BLA). Here, we used the GABAA agonist, muscimol (Mus), to inactivate BLA prior to CR and examined the effects on sleep, freezing and stress-induced hyperthermia (SIH). Wistar rats (n=28) were implanted with electrodes for recording sleep, data loggers for recording core body temperature, and with cannulae aimed bilaterally into BLA. After recovery, the animals were habituated to the injection procedure and baseline sleep was recorded. On experimental day 1, rats received ST (20 footshocks, 0.8mA, 0.5s duration, 60s interstimulus interval). On experimental day 7, the rats received microinjections (0.5ul) into BLA of either Mus (1.0uM; n = 13) or vehicle (Veh; n = 15) prior to CR (CR1). On experimental day 21, the animals experienced a second CR (CR2) without Mus. For analysis, the rats were separated into 4 groups: (Veh-vulnerable (Veh-Vul; n=8), Veh-resilient (Veh-Res; n=7), Mus-vulnerable (Mus-Vul; n=7), and Mus-resilient (Mus-Res; n=6)) based on whether or not REM was decreased, compared to baseline, during the first 4 h following ST. Pre-CR1 inactivation of BLA did not alter freezing or SIH, but did block the reduction in REM in the Mus-Vul group compared to the Veh-Vul group. These data indicate that BLA is an important region for mediating the effects of fearful memories on sleep.

Study Objectives Disturbed overnight sleep is a prominent feature of advanced stage Huntington’s disease (HD). Several polysomnography (PSG) studies have reported significant changes of sleep in HD patients, but the findings are not unequivocal. To date, no meta-analysis has investigated the PSG changes in HD patients. The present study meta-analyzed results from studies examining the PSG changes in HD patients compared with controls. Methods A literature search performed in MEDLINE, EMBASE, All EBM databases, PsycINFO, and CINAHL databases identified seven studies involving 152 HD patients and 144 controls which were included in our meta-analysis. Results Pooled results indicated decreased sleep efficiency, percentage of slow wave sleep and rapid eye movement sleep, and increased percentage of N1 sleep, wake time after sleep onset, and rapid eye movement sleep latency in HD patients compared with controls. We found high heterogeneity in the effect sizes and no indication of systematic publication biases across studies. Meta-regression analyses showed that some of the heterogeneity was explained by age, body mass index (BMI), CAG repeat length, and disease severity of HD patients. Conclusions Our study showed that polysomnographic abnormalities are present in HD. Our findings also underscore the need for a comprehensive PSG assessment of sleep changes in patients with HD. Furthermore, the effects of age, BMI and CAG repeat length on sleep changes should be carefully considered and closely monitored in the management of HD.

Astronauts on deep space missions will be required to work autonomously and thus their ability to perform executive functions could be critical to mission success. Ground-based rodent experiments have shown that low (

Systemic administration of the Group II metabotropic glutamate (mGlu) receptor agonist, LY379268 (LY37), dose-dependently suppresses rapid eye movement sleep (REM) whereas systemic administration of the mGlu II receptor antagonist, LY341495 (LY34), increases arousal. Group II mGlu receptors are highly expressed in the amygdala, a brain region involved in the regulation of sleep and arousal. To determine whether the amygdala is involved in mediating the effects of Group II mGlu agents on sleep, we microinjected LY37 and LY34 into the basal amygdala (BA) and the central nucleus of the amygdala (CNA) and recorded sleep and wakefulness. Wistar rats were implanted with electrodes for recording sleep and with bilateral cannulae aimed into BA for drug administration. Different groups of rats received bilateral microinjections of LY37 into BA at two dosage ranges (3.2 mM, 5.3 mM or 10.7 mM or 0.1 nM, 2.0 nM or 10.0 nM) or one dosage range of LY34 (1.0 nM, 30.0 nM or 60.0 nM). Microinjections into CNA were conducted at one dosage range for LY37 (0.1 nM, 2.0 nM or 10.0 nM) and for LY34 (1.0 nM, 30.0 nM or 60.0 nM). All drugs or vehicle alone were administered in a counterbalanced order at 5-day intervals. Following microinjection, sleep was recorded for 20 h. Microinjection of LY37 into BA at both nM and mM concentrations significantly decreased REM without significantly altering NREM, total sleep or wakefulness. The high dosage of LY34 in BA significantly suppressed NREM and total sleep. Microinjections of LY37 or LY34 into CNA had no significant impact on sleep. We suggest that Group II mGlu receptors may influence specific cells in BA that control descending output (via the CNA or bed nucleus of the stria terminalis) that in turn regulates pontine REM generator regions.

Study objectives Contextual fear is followed by significant reductions in rapid eye movement sleep (REM) that are regulated by the central nucleus of the amygdala (CNA). Corticotropin-releasing factor (CRF) plays a major role in regulating the stress response as well as arousal, and CRF in CNA is implicated in stress-related behavior. To test the hypothesis that CRF regulation of CNA is involved in fear-induced alterations in REM, we determined the effects of microinjections into CNA of the CRF1 antagonist, antalarmin (ANT) on fear-induced reductions in REM. We also evaluated c-Fos activation in the hypothalamic paraventricular nucleus (PVN), locus coeruleus (LC), and dorsal raphe nucleus (DRN) to determine whether activation of these regions was consistent with their roles in regulating stress and in the control of REM. Design On separate days, rats were subjected to baseline and 2 shock training sessions (S1 and S2). Five days later, the rats received bilateral microinjections of ANT (4.8 mM) or vehicle (VEH) prior to exposure to the fearful context. Sleep was recorded for 20 h in each condition. Freezing was assessed during S1, S2, and context. Separate groups of rats received identical training and microinjections or handling control (HC) only, but were sacrificed 2 h after context exposure to assess c-Fos expression. Setting NA. Patients or participants NA. Interventions NA. Measurements and results Compared to baseline, S1 and S2 significantly reduced REM. Exposure to the fearful context reduced REM in VEH treated rats, whereas REM in ANT treated rats did not differ from baseline. ANT did not significantly alter freezing. Fear-induced c-Fos expression was decreased in PVN and LC after ANT compared to VEH. Conclusions The results demonstrate that CRF receptors in CNA are involved in fear-induced reductions in REM and neural activation (as indicated by c-Fos) in stress and REM regulatory regions, but not in fear-induced freezing.

Contextual fear significantly reduces rapid eye movement sleep (REM) during post-exposure sleep in mice and rats. Corticotropin releasing factor (CRF) plays a major role in CNS responses to stressors. We examined the influence of CRF and astressin (AST), a non-specific CRF antagonist, on sleep after contextual fear in BALB/c mice. Male mice were implanted with transmitters for recording sleep via telemetry and with a guide cannula aimed into the lateral ventricle. Recordings for vehicle and handling control were obtained after ICV microinjection of saline (SAL) followed by exposure to a novel chamber. Afterwards, the mice were subjected to shock training (20 trials, 0.5 mA, 0.5 s duration) for 2 sessions. After training, separate groups of mice received ICV microinjections of SAL (0.2 microl, n=9), CRF (0.4 microg, n=8), or AST (1.0 microg, n=8) prior to exposure to the shock context alone. Sleep was then recorded for 20 hours (8-hour light and 12-hour dark period). Compared to handling control, contextual fear significantly decreased REM during the 8-h light period in mice receiving SAL and in mice receiving CRF, but not in the mice receiving AST. Mice receiving CRF exhibited reductions in REM during the 12-h dark period after contextual fear, whereas mice receiving SAL or AST did not. CRF also reduced non-REM (NREM) delta (slow wave) amplitude in the EEG. Only mice receiving SAL prior to contextual fear exhibited significant reductions in NREM and total sleep. These findings demonstrate a role for the central CRF system in regulating alterations in sleep induced by contextual fear.

Sleep mechanisms and synaptic plasticity are thought to interact to regulate homeostasis and memory formation. However, the influences of molecules that mediate synaptic plasticity on sleep are not well understood. In this study we demonstrate that mice lacking Rab3 interacting molecule 1 alpha (RIM1 alpha) (Rim1 alpha KO), a protein of the synaptic active zone required for certain types of synaptic plasticity and learning, had 53+/-5% less baseline rapid eye movement (REM) sleep compared with their wild type littermates. Also, compared with wild type littermates, exposure of the mice to an open field or to a novel object induced more robust and longer lasting locomotion suggesting altered habituation. This difference in exploratory behavior correlated with genotype specific changes in REM and deregulated release of norepinephrine in the cortex and basal amygdala of the Rim1 alpha KO mice. Also, moderate sleep deprivation (4 h), a test of the homeostatic sleep response, induced REM sleep rebound with different time course in Rim1 alpha KO and their wild type littermates. As norepinephrine plays an important role in regulating arousal and REM sleep, our data suggest that noradrenergic deficiency in Rim1 alpha KO animals impacts exploratory behavior and sleep regulation and contributes to impairments in learning.

We examined individual differences in sleep and motor activity across two consecutive days in rats. EEG and motor activity were recorded via telemetry in Wistar rats (n=29) for 48 h under well-habituated conditions. Rats were grouped based on sleep amounts and stability across days (short [SS, n=7], intermediate [IS, n=15] and long [LS, n=7] sleep) and comparisons were conducted to determine group differences for measures of sleep and motor activity. We found that correlations across recording days were significant for all selected sleep measures and motor activity counts. Rankings for 24 h total sleep time and non-rapid eye movement sleep (NREM) were SS < IS < LS rats whereas amounts of rapid eye movement sleep did not differ among groups. Further analyses of NREM episode parameters found significant differences in mean episode length (SS < IS < LS) but not in the number of episodes. Total and average motor activity counts (per waking min) were greater (32-38%) in SS compared to LS rats on both recording days. The results indicate that individual differences in sleep and motor activity in Wistar rats are stable across days. Differences between SS and LS rats have parallels to those reported for short and long sleep humans.

Studies using various methodologies have implicated n. reticularis pontis oralis (RPO) and n. subcoeruleus (SubC) in the generation of rapid eye movement sleep (REM). In rats, electrolytic lesions in these regions may give rise to the phenomenon of REM without atonia (REM-A), in which the electrophysiological features of REM are normal except that atonia is absent and elaborate behaviors may be exhibited. However, electrolytic lesions damage both cell bodies and fibers of passage, and the neural reorganization and adaptation that can occur post-lesion can complicate interpretation. Tetrodotoxin (TTX) is a sodium channel blocker that temporarily inactivates both neurons and fibers of passage and thus may be functionally equivalent to an electrolytic lesion, but without allowing time for neural adaptation. In this study, we examined the influence of microinjections of TTX into RPO and SubC on sleep in freely behaving rats. Rats (90 day old male Sprague-Dawley) were implanted with electrodes for recording EEG and EMG. Guide cannulae were implanted aimed into RPO or SubC. Each animal received one unilateral microinjection (TTXUH: 5.0 ng/0.2 microl) and two bilateral microinjections (TTXBL: 2.5 ng/0.1 microl; TTXBH: 5.0 ng/0.2 microl) of TTX, and control microinjections of saline alone (SAL). The injections were made 2 h following lights on, and sleep was recorded for the subsequent 22 h. Sleep was scored from computerized records in 10 s epochs. Recordings from the 10-h light period and the 12-h dark period were examined separately. TTX inactivation of RPO could decrease REM and non-REM (NREM), whereas inactivation of SubC produced relatively more specific decreases in REM with smaller effects on NREM. The results complement studies that have implicated RPO and SubC in REM generation. REM-A was not observed, suggesting that REM-A is a complex phenomenon that requires time for reorganization of the nervous system after insult.

Modafinil, a novel non-amphetamine stimulant recently approved for the treatment of narcolepsy, has been shown to increase waking in both animals and humans. However, its mechanism of action is currently unknown. Earlier research into the brain structures responsible for the wake-producing actions of modafinil implicated the central nucleus of the amygdala (ACe) as a possible site of action [Neuroscience 87 (1998) 905-911; Neurosci. Lett. 241 (1998) 95-98]. The present experiments were designed to test the hypothesis that the ACe is, at least in part, involved in the wake-producing actions of modafinil. In the first experiment, rats with lesions of the ACe were injected systemically with varying doses of modafinil and sleep was recorded. At the highest dose, modafinil significantly increased waking and decreased sleep. However, there was no interaction between the lesion and the effect of the drug. In the second experiment, varying doses of modafinil were injected directly into the ACe and sleep was recorded. Injection of modafinil into the ACe did not affect sleep architecture. Thus, ACe does not play a simple role in modafinil's wake-promoting action. We suggest that more complex testing will be required to elucidate its role.

Fractionations are 20- to 100-ms pauses in diaphragm activity that occur spontaneously during rapid-eye-movement (REM) sleep, sometimes in association with pontogeniculooccipital (PGO) waves. Auditory stimuli can elicit fractionations or PGO waves during REM sleep, non-REM (NREM) sleep, and waking; however, their interrelationship has not been investigated. To determine whether the two phenomena are produced by a common phasic-event generator in REM sleep, we examined PGO waves and fractionations that were elicited by auditory stimuli (tones) presented to freely behaving cats across states. Tones elicited PGO waves and two types of fractionations: short-latency fractionation responses (SFRs; 10- to 60-ms latencies) and long-latency fractionation responses (LFRs; 60- to 120-ms latencies). Both a PGO wave and a SFR were elicited in 60–70% of trials across states, but each could be elicited alone. The latencies and durations of elicited SFRs were similar across states, but the latencies of elicited PGO waves in REM sleep (mean 62.5 ms) were significantly longer than in waking or NREM sleep. Elicited SFRs consistently occur with shorter latencies than do PGO waves, in contrast to spontaneous fractionations, which have a variable relationship to PGO waves and usually occur 10–40 ms after the onset of the PGO wave. The LFR then, elicited most frequently during REM sleep, resembles a spontaneous fractionation in its temporal relationship to the PGO wave and may reflect the bias toward motoneuronal inhibition characterizing REM sleep but not NREM sleep or waking. We conclude that, although PGO waves and SFRs share some features, like LFRs they probably are generated by different neuronal populations. In three cats there was no correlation between PGO waves and fractionations, whereas in one cat they were associated in REM sleep (LFRs and SFRs) and waking (SFRs only). Thus the majority of evidence argues against the existence of a common phasic-event generator in REM sleep.

In contrast to the many neural studies into the mechanisms of sleep onset and maintenance, few studies have focused specifically on awakening from sleep. However, the abrupt electrographic changes and large brief cardio-respiratory activation at awakening suggest that a distinct, transiently aroused, awake state may exist compared to later wakefulness. To test this hypothesis we utilized the acoustic startle reflex, a standard un-conditioned reflex elicited by a sudden loud noise. This reflex is modulated under specific conditions, one being a diminution of startle when a quieter pre-stimulus is presented immediately before the loud stimulus. This pre-pulse inhibition (PPI) is used as a measure of sensorimotor gating, with smaller PPI indicating less filtering of sensory inputs and increased responsiveness to external stimuli. Eight rats with electrodes for recording sleep-wake state were studied. An accelerometer measured startle responses. The startle reflex was elicited by 115 dB, 40 ms tones. PPI was produced by 74 dB, 20 ms tones preceding the 115 dB tone by 100 ms. Responses within 100 ms were measured. Stimuli were applied either 3-10 s after spontaneous awakenings, or in established wakefulness (30 s). Responses to the startle stimuli alone were similar in the different awake states (P = 0.821). However, PPI was smaller at awakening from non-REM sleep compared to established wakefulness (45.4 +/- 7.5% vs. 74.3 +/- 6.1%, P = 0.0002). PPI after awakening from REM sleep (52.8 +/- 17.9%) was not significantly different than established wakefulness (P = 0.297). Reduced PPI of the startle reflex at awakening from non-REM sleep supports the hypothesis that wakefulness immediately after spontaneous sleep episodes is neurophysiologically distinct from later wakefulness and associated with reduced gating of motor responses to sensory inputs. Spontaneous activation of this distinct, transiently aroused, state upon awakening may serve a protective function, preparing an animal to respond immediately to potentially threatening stimuli.

Intranasal application of vesicular stomatitis virus (VSV) produces a well-characterized model of viral encephalitis in mice. Within one day post-infection (PI), VSV travels to the olfactory bulb and, over the course of 7 days, it infects regions and tracts extending into the brainstem followed by clearance and recovery in most mice by PI day 14 (PI 14). Infectious diseases are commonly accompanied by excessive sleepiness; thus, sleep is considered a component of the acute phase response to infection. In this project, we studied the relationship between sleep and VSV infection using C57BL/6 (B6) and BALB/c mice. Mice were implanted with transmitters for recording EEG, activity and temperature by telemetry. After uninterrupted baseline recordings were collected for 2 days, each animal was infected intranasally with a single low dose of VSV (5 × 104 PFU). Sleep was recorded for 15 consecutive days and analyzed on PI 0, 1, 3, 5, 7, 10, and 14. Compared to baseline, amounts of non-rapid eye movement sleep (NREM) were increased in B6 mice during the dark period of PI 1–5, whereas rapid eye movement sleep (REM) was significantly reduced during the light periods of PI 0–14. In contrast, BALB/c mice showed significantly fewer changes in NREM and REM. These data demonstrate sleep architecture is differentially altered in these mouse strains and suggests that, in B6 mice, VSV can alter sleep before virus progresses into brain regions that control sleep.

Fear conditioning [inescapable shock training (ST)] and fearful context re-exposure (CR) alone can produce significant fear indicated by increased freezing and reductions in subsequent rapid eye movement (REM) sleep. Damage to or inactivation of the basolateral nucleus of the amygdala (BLA) prior to or after ST or prior to CR generally has been found to attenuate freezing in the shock training context. However, no one has examined the impact of BLA inactivation on fear-induced changes in sleep. Here, we used the GABAA agonist, muscimol (MUS), to inactivate BLA prior to ST, the period when fear is learned, and assessed sleep after ST and sleep and freezing after two CR sessions. Wistar rats (n = 14) were implanted with electrodes for recording sleep and with cannulae aimed bilaterally into BLA. After recovery, the animals were habituated to the injection procedure (handling) over 2 consecutive days and baseline sleep following handling was recorded. On experimental day 1, the rats were injected (0.5 μl) into BLA with either MUS (1.0 μM; n = 7) or vehicle (distilled water, n = 7) 30 min prior to ST (20 footshocks, 0.8 mA, 0.5-s duration, 60-s interstimulus interval). On experimental days 7 and 21, the animals experienced CR (CR1 and CR2, respectively) alone. Electroencephalogram and electromyogram were recorded for 8 h on each day, and the recording was scored for non-rapid eye movement sleep, REM sleep, and wakefulness. Freezing was examined during CR1 and CR2. MUS microinjections into BLA prior to ST blocked the post-training reduction in REM sleep seen in vehicle-treated rats. Furthermore, in MUS-treated rats, REM sleep after CR1 and CR2 was at baseline levels and freezing was significantly attenuated. Thus, BLA inactivation prior to ST blocks the effects of footshock stress on sleep and reduces fear memory, as indicated by the lack of freezing and changes in sleep after CR. These data indicate that BLA is an important regulator of stress-induced alterations in sleep and an important site for forming fear memories that can alter sleep.

Study objective To determine whether corticotropin-releasing factor (CRF) in the basolateral amygdala (BLA) modulated sleep and fear-conditioned alterations in sleep. Design After 2 days of habituation to recording procedures, baseline sleep recordings were obtained. The animals were then habituated to the handling procedure necessary for microinjections over 2 consecutive days. In experiment 1, rats received microinjections of 0.5 μL antalarmin (1.61 or 4.82 mM), a CRF receptor 1 antagonist, or distilled water once a week for 3 wk. In experiment 2, rats received a microinjection of either antalarmin or vehicle prior to inescapable shock training (ST; 20 shocks; 0.8 mA, 0.5 sec; 1 min interstimulus interval). The animals were placed back in the context 7 days later for 30 min without shock (CR; context re-exposure). Sleep was recorded for 8 h after each manipulation. Setting NA. Subjects Outbred Wistar rats. Interventions The rats were surgically implanted with electrodes for recording the electroencephalogram and electromyogram for determining arousal state and with bilateral guide cannulae directed at BLA. Measurements and results Antalarmin microinjected into BLA did not significantly alter sleep under undisturbed conditions. However, antalarmin microinjected bilaterally into BLA prior to ST blocked reductions in rapid eye movement sleep that ST normally produces. Further, the single microinjection prior to ST blocked the reduction in rapid eye movement typically seen after subsequent CR. Behavioral freezing, an indicator of fear memory, was not altered. Conclusions CRF in BLA is involved in regulating stress-induced alterations in sleep and it plays a role in modulating how stressful memories influence sleep.

Study Objectives: Predictability and controllability are important factors in the persisting effects of stress. We trained mice with signaled, escapable shock (SES) and with signaled, inescapable shock (SIS) to determine whether shock predictability can be a significant factor in the effects of stress on sleep. Design: Male BALB/cJ mice were implanted with transmitters for recording EEG, activity, and temperature via telemetry. After recovery from surgery, baseline sleep recordings were obtained for 2 days. The mice were then randomly assigned to SES (n = 9) and yoked SIS (n = 9) conditions. The mice were presented cues (90 dB, 2 kHz tones) that started 5.0 sec prior to and co-terminated with footshocks (0.5 mA; 5.0 sec maximum duration). SES mice always received shock but could terminate it by moving to the non-occupied chamber in a shuttlebox. SIS mice received identical tones and shocks, but could not alter shock duration. Twenty cue-shock pairings (1.0-min interstimulus intervals) were presented on 2 days (ST1 and ST2). Seven days after ST2, SES and SIS mice, in their home cages, were presented with cues identical to those presented during ST1 and ST2. Setting: NA. Patients or Participants: NA. Interventions: NA. Measurements and Results: On each training and test day, EEG, activity and temperature were recorded for 20 hours. Freezing was scored in response to the cue alone. Compared to SIS mice, SES mice showed significantly increased REM after ST1 and ST2. Compared to SES mice, SIS mice showed significantly increased NREM after ST1 and ST2. Both groups showed reduced REM in response to cue presentation alone. Both groups showed similar stress-induced increases in temperature and freezing in response to the cue alone. Conclusions: These findings indicate that predictability (modeled by signaled shock) can play a significant role in the effects of stress on sleep.

Ponto-geniculo-occipital (PGO) waves spontaneously occur in the pons, lateral geniculate body (LGB), and occipital cortex during rapid eye movement sleep (REM), and PGO-like waves (PGOE) may be elicited in LGB during sleep and waking. Because REM has been hypothesized to be a state of continual "orienting" or "hyper-alertness," we tested whether the amplitudes of PGOE in "alerting" situations (the abrupt onset of a loud sound or presentation of a novel stimulus within a series of stimuli) that evoke orienting responses (OR) would be greater than those following stimuli without OR. We also compared PGOE accompanying OR to PGOE during REM and NREM when OR are absent. The amplitudes of PGOE in W were greatest when OR were observed, and the amplitudes of PGOE accompanying OR were not significantly different from PGOE amplitudes in REM. Likewise, the amplitudes of PGOE during REM were not significantly different from those of the highest amplitude spontaneous PGO waves. We propose that the presence of PGOE signals registration of stimuli and that stimuli of sufficient significance to induce behavioral OR in waking also elicit PGOE of significantly greater amplitudes in all behavioral states. These findings support the hypothesis that the presence of high-amplitude PGO waves in REM indicates that the brain is in a state of more-or-less continual orienting.

STRESS CAN HAVE A SIGNIFICANT, LONG-LASTING NEGATIVE IMPACT ON HEALTH INCLUDING CHANGES IN BEHAVIOR AND SLEEP. HOWEVER, STRESSORS are often encountered without producing permanent or pathological changes. The difference between successful and unsuccessful coping with stress may involve the resilience of the organism1 as well as characteristics of the stressor. Stressor controllability, along with intensity and predictability, have been found to be important factors in the effects of stress.2 For example, lack of stressor controllability has been suggested to be a factor in the development of posttraumatic stress disorder (PTSD).3,4 Conditioned fear training is conducted with a fear-inducing stressor (usually inescapable shock, [IS])5,6 presented in an experimental paradigm in which the animal receiving training has no control over the stressor. Through association to IS, initially neutral environmental cues and contexts acquire the capacity to elicit behavioral and physiological responses indicative of fear and anxiety, including behavioral freezing,7–9 autonomic responses,10–12 and fear-potentiated startle.5,6 Disturbances in sleep often follow a stressful or traumatic event,13 and we and others have demonstrated that IS and IS-associated fearful cues and contexts produce similar alterations in sleep, including a prominent reduction in rapid eye movement sleep (REM).14,15 By comparison, training with avoidable, signaled shock (a controllable stressor) in a shuttlebox is followed by significant increases in REM,15–18 even though the same stressor (footshock) is initially experienced before the avoidance response is acquired. The results of these studies have typically been focused on demonstrating a correlation between REM and acquisition of the avoidance response.15–18 It is possible to train animals with escapable shock (ES) as a controllable stressor using a shuttlebox paradigm. In this paradigm, the stressor is always experienced but can be terminated by the animal's action. The effects of ES and IS have not been directly compared with respect to post-stress alterations in sleep. Therefore, in this study, we used a yoked design to examine the effects of ES and IS training and of contextual reminders of ES and IS on sleep in BALB/cJ mice, a “behaviorally reactive” strain that shows greater post-stress reductions in REM and alterations in sleep in response to IS compared to C57BL/6J mice.19–21 We also compared mice in the ES and IS conditions for potential differences in gross motor activity and we examined freezing, a common behavioral index of fear,7,22,23 during exposure to the fearful context. Examining freezing enabled us to compare immediate emotional reactivity across conditions and to determine the relationship of initial emotionality to subsequent changes in arousal and sleep.

The amygdala has emerged as an important forebrain modulator of arousal. Acetylcholine plays a role in the regulation of sleep and wakefulness, particularly rapid eye movement sleep (REM). The major cholinergic input to the amygdala comes from the basal forebrain, a region primarily linked to wakefulness. We examined sleep and the encephalogram for 8 h following bilateral microinjections into the central nucleus of the amygdala (CNA) of the cholinergic agonist, carbachol (CARB(L): 0.3 microg; CARB(H): 3.0 microg), the acetylcholinesterase inhibitor, neostigmine (NEO(L): 0.3 microg; NEO(H): 3.0 microg), the muscarinic antagonist, scopolamine (SCO(L): 0.3 microg; SCO(H): 1.0 microg), the nicotinic antagonist, mecamylamine (MEC(L): 0.3 microg; MEC(H): 1.0 microg) and saline (SAL, 0.2 microl) alone. Both doses of CARB and NEO significantly reduced REM, but did not significantly alter non-rapid eye movement sleep (NREM). Both doses of SCO significantly increased NREM, and SCO(H) also produced an initial increase in REM followed by a significant decrease. CARB(H) and NEO(H) decreased REM electroencephalogram (EEG) power in the 5.5-10 Hz band, and NEO(L) and NEO(H) decreased NREM EEG power in the 0.5-5.0 Hz band. CARB(L) decreased waking EEG power in the 0.5-5.0 Hz band, and NEO(H) decreased waking EEG power in the 5.0-10.0 Hz band. Both doses of SCO significantly increased waking EEG power in the 5.5-10.0 Hz band. Compared with SAL, MEC did not significantly alter sleep or EEG power. The reduction of REM by CARB and NEO and the alteration of sleep by SCO indicate that cholinergic regulation of the amygdala is involved in the control of arousal in rodents. In contrast, CARB microinjections into CNA increase REM in cats, though the reasons for the species difference are not known. The results are discussed in the context of anatomical inputs and species differences in the cholinergic regulation of CNA.

Study objectives To determine the effects of temporary functional inactivation of the central nucleus of the amygdala on sleep and on activity in an arousing environment, an open field. Design Rats were implanted with electrodes for recording the electroencephalogram (EEG) and electromyogram (EMG), and with guide cannulae aimed into CNA. Sleep was recorded for 22 h (10 h light, 12 h dark) following microinjections of tetrodotoxin (TTX: 5.0 ng/0.2 microl given unilaterally [TTXUH] or bilaterally [TTXBH], and 2.5 ng/0.1 microl given bilaterally [TTXBL]) or saline (SAL) alone on separate days. Activity during 1 h in an OF was recorded after microinjections of TTXBH and SAL. Setting NA. Patients or participants Three-month-old Wistar rats (n=12). Interventions Functional inactivation of the central nucleus of the amygdala with TTX. Measurements and results Compared to SAL, all TTX microinjections significantly shortened NREM latency, but did not alter total NREM during either light or dark periods. During the light period, TTXBH significantly reduced total REM and REM episode number, and TTXBL decreased REM episode number. All TTX microinjections increased EEG slow wave activity (0.5-4 Hz, SWA) during wakefulness, NREM and REM. Activity in OF was decreased after TTXBH compared to SAL. Conclusions Functional lesions of the amygdala, including the central nucleus of the amygdala, decreased REM sleep and reduced arousal, as indicated by shortened NREM latency and decreased activity in an arousing environment. These findings suggest that the amygdala plays a broad role in modulating spontaneous sleep and wakefulness and in modulating responsiveness in arousing situations.

The amygdala has been implicated in emotional arousal and in the regulation of sleep. Previously, we demonstrated that tetrodotoxin (TTX), a sodium channel blocker that temporarily inactivates neurons and tracts, microinjected into the central nucleus of the amygdala (CNA) during the light period significantly reduced REM, shortened sleep latency, and increased EEG delta power in rats. TTX inactivation of CNA also reduced activity in the open field. These findings suggest that the amygdala modulates arousal in a variety of situations. To test the hypothesis that the amygdala may influence spontaneous arousal, we examined the effects of TTX inactivation of CNA on sleep and activity during the dark period when rats show higher arousal and less sleep. EEG and activity were recorded via telemetry in Wistar rats (n = 8). Bilateral microinjections of TTX (L: 2.5 ng/0.1; H: 5.0 ng/0.2 microl) or SAL (saline, 0.2 microl) were administered before lights off followed by recording throughout the 12-h dark period and following 12-h light period. Microinjections were given at 5-day intervals and were counterbalanced across condition. TTX significantly shortened sleep latency, increased NREM time, decreased REM time, and decreased activity. TTX increased NREM episode duration, whereas the number and duration of REM episodes were decreased. The present results indicate that TTX inactivation of CNA can increase NREM time when spontaneous arousal is high, suggesting a broad role for the amygdala in regulating arousal. The results suggest that understanding the ways in which the amygdala modulates arousal may provide insight into the mechanisms underlying altered sleep in mood and anxiety disorders.

Genetic and physiological diversity amongst rodent strains provide the potential for developing models that may give insight into factors that regulate sleep in response to environmental challenges. We examined home cage activity, behavioral performance in the open field and sleep after a number of mild stressors (cage change [CC], open field [OF]) and after 1 and 4h of sleep deprivation (1hSD and 4hSD) in rat strains (Fischer 344 [F344], Lewis [LEW], Wistar [WST] and Sprague-Dawley [Sp-D], n=16 per strain) that differ in behavior and sleep. F344 and WST rats had greater home cage locomotion than LEW and Sp-D rats, but F344 rats exhibited the least relative locomotion in OF. In 24h baseline recordings of sleep, strain rankings were LEW=WST=Sp-D>F344 in rapid eye movement sleep (REM), and LEW=Sp-D>F344 and LEW>WST in non-REM (NREM). Compared to baseline, total sleep was reduced in all four strains after CC, OF and 1hSD, but not after 4hSD, in the first hour after treatment. Afterwards, increases in REM and NREM were seen after all treatments with the amount and time course varying across treatments and strains. CC induced the weakest and 4hSD the largest effects on sleep, whereas OF and 1hSD had intermediate effects. Among strains, the more anxious F344 rats exhibited the greatest sleep increases during the light period after OF, 1hSD and 4hSD. The results are discussed with respect to the relationship between behavioral and sleep responses to stressors, and to potential mechanisms underlying the strain differences.

Shock training and auditory cues associated with shock produce alterations in sleep that can be long-lasting in BALB/cJ (C) mice. We examined sleep in C mice after different amounts of shock training, and after cues with different strength cue-shock associations. Mice were implanted with transmitters for determining sleep via telemetry. After baseline sleep recording, the mice were trained (between 08:00 and 09:00 h) to associate a cue (tone) with footshock in either single shock training (SST: a single tone-shock pairing) or multiple shock training (MST: 15 tone-shock pairings) conditions. For testing, the mice were presented 15 cues (tone only) in their home cage between 10:45 and 11:00 h on post-training days 6, 13, 20, 27, and 34 (Cue 1 to Cue 5) following shock training. Sleep was recorded for two days after shock training or cue presentation. A separate group of mice received 15 tone-shock pairings and had their sleep recorded for 10 consecutive uninterrupted days. Both SST and MST mice showed decreases in rapid eye movement sleep (REM) after shock training, with the larger effect in the MST mice. Only MST mice showed significant reductions in REM in response to the fearful cues, and longer-term alterations in sleep could be observed even on the day after cue presentation. These results indicate that reminders of an aversive event can impact sleep for prolonged periods, and that the degree of the impact varies with amount of training.

Fear conditioning is thought to model anticipatory anxiety. Inbred mouse strains exhibit different levels of reactivity to aversive environmental stimuli, which may reflect anxiety. We examined the effects of fear conditioning on sleep in mouse strains that differ on behavioral measures of anxiety. Mice (BALB/cJ [C], C57BL/6J [B6], CB6F1/J [CB6], n = 7-10 per strain) were implanted with transmitters for recording sleep by telemetry. Baseline sleep was recorded, and the mice were trained to associate a cue (tone) with footshock (15 cue-shock pairings on 4 consecutive days). Sleep was recorded after shock training and again 4 to 5 days later after presentation of the cue alone. Shock training produced a relatively selective suppression of rapid eye movement sleep (REM) that was greater in the C strain compared to the B6 and CB6 mice. Post-training exposure to the cue alone suppressed REM in all strains. The C strain exhibited a relatively greater immediate suppression of REM, and the CB6 hybrid mice showed the greatest overall suppression of REM. These data demonstrate that stimuli associated with an aversive event can alter sleep and suppress REM in much the same way as exposure to the event itself. Fear conditioning may provide a model for examining genetic and neural mechanisms underlying the influence of anxiety on sleep.

It is becoming established that the amygdala has a strong influence on arousal state, with most evidence indicating a role in the regulation of rapid eye movement sleep (REM). Electrically activating the central nucleus of the amygdala (CNA) can increase subsequent REM and enhance REM-related phenomena. However, drugs that may be inhibitory to CNA have been typically reported to reduce REM. This suggests that enhancing activity in CNA could promote REM, and that inhibiting activity in CNA could suppress REM. We reversibly inactivated CNA using the GABA(A) agonist, muscimol, or blocked GABAergic inhibition with the GABA(A) antagonist, bicuculline, and examined the effects on sleep and wakefulness. Rats (90-day-old male Sprague-Dawley) were implanted with electrodes for recording EEG and EMG. Cannulae were aimed into CNA for microinjecting muscimol (0.001, 0.3 and 1.0 microM/0.2 microl saline) or bicuculline (56 and 333 pM/0.2 microl saline). Each animal received bilateral microinjections of muscimol, bicuculine or saline alone followed by 6-h sleep recordings. Microinjections of low concentrations of muscimol into CNA produced relatively selective decreases in total REM and number of REM episodes that lasted up to 6 h. In contrast, microinjections of bicuculline into CNA produced significant increases in REM. There were no significant reductions in NREM or wakefulness. These findings demonstrate that inactivating CNA can produce a relatively selective suppression of REM. The possible role that spontaneous activity in CNA may play in REM initiation and/or maintenance is discussed.

This study assessed differences in spontaneous sleep and locomotor activity in inbred and hybrid mouse strains and evaluated telemetry for recording sleep in mice.Uninterrupted recordings of sleep and home cage activity were obtained in four mouse strains. Pre-operative and post-operative home cage activity was obtained in two strains.N/A PARTICIPANTS: The subjects were mice of three inbred (C57BL/6J (B6), n=25; BALB/cJ (C), n=24; DBA/2J (D2), n=30) strains and one hybrid (CB6F1/J (CB6: C x B6), n=19) strain.Electroencephalogram (EEG) and activity were recorded by telemetry, and behavioral states were visually scored based on EEG and activity records. Home cage activity was determined utilizing photobeam interruptions.1) Among the three inbred strains: C mice had the least sleep and the greatest amount of activity; D2 mice exhibited the least non-rapid-eye-movement (NREM) sleep, the longest average NREM-bout length, and the greatest diurnal ratio of sleep and were the most inactive; B6 mice had the most sleep and an intermediate activity level; no differences among inbred strains were observed in total REM. The CB6 mice exhibited intermediate levels of total sleep and activity and had greater amounts of REM compared to its parental strains. 2) Total operative mortality was 9.3%, with all deaths occurring within 3 to 9 days after the operation; significant reductions in activity were observed after the operation.Differences in spontaneous sleep and activity exist among inbred and hybrid mouse strains. Accurate determination of sleep states in mice can be achieved with telemetrically recorded EEG and activity.

The influence of external stimuli and the memories of both unpleasant and pleasant conditions clearly can have a considerable impact on the quality of sleep. The amygdala, a structure that plays an important role in coding the emotional significance of stimuli and is heavily interconnected with brainstem nuclei known to be involved in sleep control, has received little attention from sleep researchers. We report on a series of studies, focusing on its central nucleus (Ace). Presence of serotonin (5-HT) in Ace caused a rapid change of state when injected in rapid- eye-movement sleep (REM) compared with non-REM (NREM) injections. A 5-HT antagonist released ponto-geniculo-occipital waves (PGO) into NREM. Stimuli conditioned by pairing with aversive stimuli in a fear-conditioning paradigm significantly increased sound-elicited PGO and reduced REM.

Waves similar to spontaneous ponto-geniculo-occipital (PGO) waves of paradoxical sleep (PS) in cats are elicited by tones and can be considered a form of evoked potential termed PGOE. The aims of the present experiment were to determine: (1) the effects of tone intensity on the probability of producing PGOE; (2) the effects of intensity on the amplitude and latency of PGOE across slow-wave sleep (SWS) and PS; (3) whether the effects of intensity on PGOE are similar to those on a particular form of auditory evoked potential known as the mid-latency response (MLR). Increasing the intensity of the stimulus from 60 to 100 in 10 dB increments resulted in increased probability, increased amplitude, and decreased latency of PGOE in both SWS and PS. This pattern was similar to published findings with MLR, and latencies of PGOE (roughly 60-100 msec) fell within the range of 'C wave' type of MLR recorded in intralaminar nuclei of the thalamus of cats. The possibility that PGOE and MLR share underlying mechanisms and represent the same phenomenon is discussed with particular attention to the function of the mechanisms during alerting and orienting.