Search

[Display omitted] • Wide- and narrow-spiking waveforms are identified in the region of locus coeruleus. • Wide-spiking units show higher excitability under anesthesia. • A subset of narrow-spiking units shows fast-spiking phenotype in wakefulness. • In-situ hybridization reveals the existence of GABAergic neurons in locus coeruleus. Neurons in the locus coeruleus (LC) have been traditionally viewed as a homogenous population. Recent studies begin to reveal their heterogeneity at multiple levels, ranging from molecular compositions to projection targets. To further uncover variations of neuronal properties in the LC, we took a genetic-based tagging approach to identify these neurons. Our data revealed diverse spike waveforms among neurons in the LC region, including a considerable fraction of narrow-spiking units. While all wide-spiking units possessed the regular waveform polarity (negative-positive deflection), the narrow units can be further divided based on opposing waveform polarities. Under anesthesia, wide units emitted action potential at a higher rate than the narrow units. Under wakefulness, only one subtype of narrow units exhibited fast-spiking phenotype. These neurons also had long latencies to optogenetic stimulation. In-situ hybridization further supported the existence of a small population of putative GABAergic neurons in the LC core. Together, our data reveal characteristic differences among neurons in the LC region, and suggest that a fraction of electrophysiologically-identified narrow-spiking neurons can be fast-spiking interneurons, and their fast-spiking feature is masked by anesthesia. [ABSTRACT FROM AUTHOR]

• Oscillatory markers of sleep need are atheoretical; they vary with time awake but cannot inform on the nature of sleep need. • Increasing sleep need relates to excitability in neuronal populations, as may be measured via aperiodic EEG activity. • We discuss the use of aperiodic EEG in measuring sleep need, as well as other issues in using EEG to measure sleep need. Sleep is a daily experience across humans and other species, yet our understanding of how and why we sleep is presently incomplete. This is particularly prevalent in research examining the neurophysiological measurement of sleepiness in humans, where several electroencephalogram (EEG) phenomena have been linked with prolonged wakefulness. This leaves researchers without a solid basis for the measurement of homeostatic sleep need and complicates our understanding of the nature of sleep. Recent theoretical and technical advances may allow for a greater understanding of the neurobiological basis of homeostatic sleep need: this may result from increases in neuronal excitability and shifts in excitation/inhibition balance in neuronal circuits and can potentially be directly measured via the aperiodic component of the EEG. Here, we review the literature on EEG-derived markers of sleepiness in humans and argue that changes in these electrophysiological markers may actually result from neuronal activity represented by changes in aperiodic markers. We argue for the use of aperiodic markers derived from the EEG in predicting sleepiness and suggest areas for future research based on these. [ABSTRACT FROM AUTHOR]

Total sleep deprivation (TSD) negatively affects cognitive functions, especially vigilance attention, but studies on vigilance changes in terms of electroencephalography (EEG) microstates after TSD are limited. This study investigates the impact of TSD on vigilance attention, EEG microstates and its relationship. Thirty healthy adult males completed a psychomotor vigilance task (PVT) before, 24 h after, and 36 h after TSD while their EEG was recorded during rest. Microstate analysis revealed significant changes in the occurrence and contribution of microstate class B after TSD. Moreover, changes in the probability of transitioning between microstate classes A and D were observed, correlating with decreased vigilance. Specifically, a positive correlation was found between transitioning from class B to class C and vigilance, while a trend of negative correlation was observed between transitioning between classes A and D and vigilance. These findings indicate abnormal activity in the salience network and dorsal attention network following sleep deprivation. TSD impairs vigilance attention, as demonstrated by the effects on EEG microstate class B and the transitions between classes A and D. The study suggests its potential as an early warning indicator for predicting vigilance attention after sleep deprivation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

[Display omitted] • Short-wavelength light during nighttime and over a long period (∼6h) can sustain inducing alertness, while long-wavelength (red) light at a shorter duration (∼1h) and lower intensity can almost produce this effect during the daytime until night. • EEG measurements of alertness reveal high activity at different times during exposure to short-wavelength and long-wavelength light, and this increase does not seem to associate with improving subjective and objective measures of alertness. • Rather than melanopic illuminance, L-cone-opic illuminance could be used to determine particular alerting effects of light, such as neural activity in the brain at specified times. • For producing alertness during nighttime, it is recommended to substitute light sources enriched at short-wavelength and middle-wavelength (420–555 nm) with long-wavelength (620 – 640 nm) light to avoid disrupting melatonin secretion. Light is detected in the eye by three classes of photoreceptors (rods, cones, and intrinsically photosensitive retinal ganglion cells (ipRGCs)) that are each optimized for a specific function and express a particular light-detecting photopigment. The significant role of short-wavelength light and ipRGCs in improving alertness has been well-established; however, few reviews have been undertaken to assess the other wavelengths' effects regarding timing and intensity. This study aims to evaluate the impact of different narrowband light wavelengths on subjective and objective alertness among the 36 studies included in this systematic review, 17 of which were meta-analyzed. Short-wavelength light (∼460–480 nm) significantly improves subjective alertness, cognitive function, and neurological brain activities at night, even for a sustained period (∼6h) (for λmax: 470/475 nm, 0.4 < |Hedges's g| < 0.6, p < 0.05), but except early morning, it almost does not show this effect during the day when melatonin level is lowest. Long-wavelength light (∼600–640 nm) has little effect at night, but significantly increases several measures of alertness at lower irradiance during the daytime (∼1h), particularly when there is homeostatic sleep drive (for λmax: ∼630 nm, 0.5 < |Hedges's g| < 0.8, p < 0.05). The results further suggest that melanopic illuminance may not always be sufficient to measure the alerting effect of light. [ABSTRACT FROM AUTHOR]

Highlights • Item memory was equally retained across early morning sleep and wakefulness. • Early morning nap sleep benefitted emotional and neutral associative recognition. • Post-sleep neutral memory performance was correlated with spindle density. • Post-sleep emotional memory performance was not correlated with spindle density. • A similar pattern of correlations was evident at delayed testing (+24 h). Abstract Emotional events are preferentially retained in episodic memory. This effect is commonly attributed to enhanced consolidation and has been linked specifically to rapid eye movement (REM) sleep physiology. While several studies have demonstrated an enhancing effect of REM sleep on emotional item memory, it has not been thoroughly explored whether this effect extends to the retention of associative memory. Moreover, it is unclear how non-rapid eye movement (NREM) sleep contributes to these effects. The present study thus examined associative recognition of emotional and non-emotional material across an early morning nap (N = 23) and sustained wakefulness (N = 23). Nap group subjects demonstrated enhanced post-sleep associative memory performance, which was evident across both valence categories. Subsequent analyses revealed significant correlations between NREM spindle density and pre-sleep memory performance. Moreover, NREM spindle density was positively correlated with post-sleep neutral associative memory performance but not with post-sleep emotional associative memory. Accordingly, only neutral associative memory, but not emotional associative memory, was significantly correlated with spindle density after an additional night of sleep (+24 h). These results illustrate a temporally persistent relationship between spindle density and memory for neutral associations, whereas post-sleep emotional associative memory appears to be disengaged from NREM-sleep-dependent processes. [ABSTRACT FROM AUTHOR]

Visual fixation (VF) of a target is a possible, although atypical, feature of the Unresponsive Wakefulness Syndrome (UWS). Whether VF may indicate residual awareness in these patients is debatable, since it may simply subtend a series of reflex processes. Objective tools should therefore be used to identify aware VF, which depends on the integrity of visuomotor networks encompassing frontal-parietal-occipital areas. The aim of our study was to detect residual visuomotor network functionality potentially sustaining aware VF. To this end, we evaluated the visuomotor integration (VMI) and visual P300 patterns in a chronic Disorder of Consciousness (DOC) sample and a control group of healthy individuals (HC), using an associative stimulation protocol combining transcranial magnetic stimulation (TMS) with visual stimulation through transorbital alternating current stimulation. The Minimally Conscious State (MCS) patients showed preserved patterns of VMI and P300, whereas nearly all the UWS patients showed no significant VMI. Notably, the electrophysiological findings were correlated with the visual domain of the Coma Recovery Scale-Revised. Nonetheless, two fixating UWS individuals had a VMI similar to MCS patients. Our data suggest that some UWS patients showing VF could be aware, but unable to manifest it clearly, probably because of a severe motor output impairment, which is a condition compatible with the Functional Locked-In Syndrome. [ABSTRACT FROM AUTHOR]

Behavioral responsiveness and awareness levels correlate with the degree of functional connectivity within cortical-thalamocortical networks, whose breakdown accounts for chronic disorders of consciousness (DOC). Our study was aimed at assessing the role of the primary motor area (M1) and premotor-M1 circuitry dysfunction in motor output deterioration in minimally conscious state (MCS) and unresponsive wakefulness syndrome (UWS) patients. As a control group, we included a healthy subject (HC) sample in the study. We evaluated the effects of different types of transcranial magnetic stimuli over M1 by recording post-stimulus time histogram (PSTH), which includes a series of peaks of unit firing activity that match with D and I-waves, characterizing the descending corticospinal volleys evoked by transcranial magnetic stimuli. As compared to HC, DOC patients showed a dysfunction of intra-M1 and premotor-M1 circuits, which correlated with the Coma Recovery Scale-Revised scorings. Nonetheless, one UWS patient showed a partially preserved premotor-M1 circuitry, paralleled by a severe intra-M1 circuitry dysfunction. Our data suggest that motor unresponsiveness in some DOC patients may be due to a pure motor output failure, as in the functional locked-in syndrome (fLIS), rather than to a premotor-motor connectivity impairment, which instead characterizes MCS and UWS. [ABSTRACT FROM AUTHOR]

The purpose of the present study was to determine whether physical exercise (PE) has a protective effect in an experimental animal model of sleep-related movement disorder (A11 dopaminergic nuclei lesions with 6-OHDA). Rats were divided into four groups (Control PE-CTRL/PE, SHAM/PE, A11 lesion/NPE, A11 lesion/PE). Two experiments were performed: (1) the rats underwent PE before (2 weeks) and after (4 weeks) the A11 lesion; and (2) the rats underwent PE only after (4 weeks) the A11 lesion. Electrode insertion surgery was performed and sleep analyses were conducted over a period of 24 h (baseline and after PE) and analyzed in 6 blocks of 4 h. The results demonstrated that the A11 lesion produced an increased percentage of wakefulness in the final block of the dark period (3–7 am) and a significant enhancement of the number of limb movements (LM) throughout the day. Four weeks of PE was important for reducing the number of LMs in the A11 lesion group in the rats that performed PE before and after the A11 lesion. However, in the analysis of the protective effect of PE on LM, the results showed that the number of LMs was lower at baseline in the group that had performed 2 weeks of PE prior to the A11 lesion than in the group that had not previously performed PE. In conclusion, these findings consistently demonstrate that non-pharmacological manipulations had a beneficial effect on the symptoms of sleep-related movement disorder. [ABSTRACT FROM AUTHOR]

Functional near infrared spectroscopy (fNIRS) is an emerging non-invasive technique that allows bedside measurement of blood oxygenation level-dependent hemodynamic signals. We aimed to examine the efficacy of resting-state fNIRS in detecting the residual functional networks in patients with disorders of consciousness (DOC). We performed resting-state fNIRS in 23 DOC patients of whom 12 were in minimally conscious state (MCS) and 11 were in unresponsive wakefulness state (UWS). Ten regions of interest (ROIs) in the prefrontal cortex (PFC) were selected: both sides of Brodmann area (BA) 9, BA10, BA44, BA45, and BA46. Graph-theoretical analysis and seed-based correlation analyses were used to investigate the network topology and the strength of pairwise connections between ROIs and channels. MCS and UWS exhibited varying degrees of the loss of topological architecture, and the regional nodal properties of BA10 were significantly different between them (Nodal degree, P Left BA10 = 0.01, P Right BA10 < 0.01; nodal efficiency, P Left BA10 = 0.03, P Right BA10 < 0.01). Compared to healthy controls, UWS had impaired functions in both short- and long-distance connectivity, however, MCS had significantly impaired functions only in long-distance connectivity. The functional connectivity of right BA10 (AUC = 0.88) and the connections between left BA46 and right BA10 (AUC = 0.86) had excellent performance in differentiating MCS and UWS. MCS and UWS have different patterns of topological architecture and short- and long-distance connectivity in PFC. Intraconnections within BA10 and interhemispheric connections between BA10 and 46 are excellent resting-state fNIRS classifiers for distinguishing between MCS and UWS., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Abstract: Vitamin A is a common lipophilic vitamin, and its function is mainly mediated by the binding of its metabolite retinoic acid to retinoic acid receptors (RARs) and retinoid X receptors. Recently, it was reported that the expression of the RARb (an RAR subtype) gene determines the contribution of the delta oscillation in the sleep electroencephalogram (EEG) patterns in mice. We also reported that 4-week dietary deficiency of vitamin A (VAD) causes the attenuation of delta power in sleep and spontaneous activity in mice. However, our previous study could not clarify whether the attenuation of delta power by VAD is attributed to the suppression of RARs. To address this problem, we investigated whether the chronic administration of LE540 (30mg/kg/day), an antagonist of RARs, for 1 or 4weeks attenuated EEG delta power during sleep in mice. Consequently, 4-week LE540 administration induced a significant attenuation of wakefulness and delta power in non-rapid eye movement sleep. Western blot analysis revealed a significant decrease in the expression of dopamine D1 receptor (D1DR) in the striatum and tyrosine hydroxylase in the midbrain of mice that were administered LE540 for 4weeks. High-performance liquid chromatography analysis of striatal tissue revealed a significant decrease in the homovanillic acid/dopamine ratio. Meanwhile, dopamine levels did not change in these mice. Our results suggest that the 4-week antagonism of RARs induces the attenuation of delta power. However, the attenuation of delta power may be elicited indirectly by the decrease of wakefulness followed by the hypo-expression of dopamine receptors especially D1DR. [Copyright &y& Elsevier]

Abstract: The microinjection of nerve growth factor (NGF) into the cat pontine tegmentum rapidly induces rapid eye movement (REM) sleep. To determine if NGF is involved in naturally-occurring REM sleep, we examined whether it is present in mesopontine cholinergic structures that promote the initiation of REM sleep, and whether the blockade of NGF production in these structures suppresses REM sleep. We found that cholinergic neurons in the cat dorso-lateral mesopontine tegmentum exhibited NGF-like immunoreactivity. In addition, the microinjection of an oligodeoxyribonucleotide (OD) directed against cat NGF mRNA into this region resulted in a reduction in the time spent in REM sleep in conjunction with an increase in the time spent in wakefulness. Sleep and wakefulness returned to baseline conditions 2 to 5days after antisense OD administration. The preceding antisense OD-induced effects occurred in conjunction with the suppression of NGF-like immunoreactivity within the site of antisense OD injection. These data support the hypothesis that NGF is involved in the modulation of naturally-occurring sleep and wakefulness. [Copyright &y& Elsevier]

Abstract: Hypothalamic neurons that utilize melanin-concentrating hormone (MCH) as a neuromodulator exert a positive control over energy homeostasis, inducing feeding and decreasing metabolism. Recent studies have shown also that this system plays a role in the generation and/or maintenance of sleep. MCHergic neurons project to the serotonergic dorsal raphe nucleus (DR), a neuroanatomical structure involved in several functions during wakefulness (W), and in the regulation of rapid-eye movements (REM) sleep. Recently, we determined the effect of MCH microinjected into the DR on sleep variables in the rat. MCH produced a marked increment of REM sleep, whereas slow wave sleep (SWS) showed only a moderate increase. In the present study, we analyze the effect of immunoneutralization of MCH in the DR on sleep and W in the rat. Compared to the control solution, microinjections of anti-MCH antibodies (1/100 solution in 0.2μl) induced a significant increase in REM sleep latency (31.2±7.1 vs. 84.2±24.8min, p <0.05) and a decrease of REM sleep time (37.8±5.4 vs. 17.8±2.9min, p <0.05) that was related to the reduction in the number of REM sleep episodes. In addition, there was an increase of total W time (49.8±4.6 vs. 72.0±5.7min, p <0.01). Light sleep and SWS remained unchanged. The intra-DR administration of a more diluted solution of anti-MCH antibodies (1/500) or rabbit pre-immune serum did not modify neither W nor REM sleep variables. Our findings strongly suggest that MCH released in the DR facilitates the occurrence of REM sleep. [ABSTRACT FROM AUTHOR]

Abstract: The hippocampal theta rhythm is associated with the processing of sensory systems such as touch, smell, vision and hearing, as well as with motor activity, the modulation of autonomic processes such as cardiac rhythm, and learning and memory processes. The discovery of temporal correlation (phase locking) between the theta rhythm and both visual and auditory neuronal activity has led us to postulate the participation of such rhythm in the temporal processing of sensory information. In addition, changes in attention can modify both the theta rhythm and the auditory and visual sensory activity. The present report tested the hypothesis that the temporal correlation between auditory neuronal discharges in the inferior colliculus central nucleus (ICc) and the hippocampal theta rhythm could be enhanced by changes in sensory stimulation. We presented chronically implanted guinea pigs with auditory stimuli that varied over time, and recorded the auditory response during wakefulness. It was observed that the stimulation shifts were capable of producing the temporal phase correlations between the theta rhythm and the ICc unit firing, and they differed depending on the stimulus change performed. Such correlations disappeared approximately 6 s after the change presentation. Furthermore, the power of the hippocampal theta rhythm increased in half of the cases presented with a stimulation change. Based on these data, we propose that the degree of correlation between the unitary activity and the hippocampal theta rhythm varies with – and therefore may signal – stimulus novelty. [Copyright &y& Elsevier]

Abstract: Activation of noradrenergic locus coeruleus (LC) neurons promotes wakefulness and behavioral arousal. In rats, LC neurons receive circadian inputs via a circuit that originates in the suprachiasmatic nucleus (SCN) and relays through the dorsomedial hypothalamus (DMH) to LC; this circuit input increases LC activity during the active period. DMH neurons expressing the peptide neurotransmitter orexin/hypocretin are ideally situated to act as a relay between SCN and LC due to their synaptic inputs from SCN and innervation of LC. Here, we examined the hypothesis that orexin is involved in transmitting circadian signals to LC using single-unit recordings of LC neurons in anesthetized rats maintained in 12:12 light–dark housing. We replicated earlier findings from this lab that LC neurons fire significantly faster on average during the active compared to rest periods. Local microinjection of an orexin antagonist, SB-334867-A attenuated the impulse activities of the fastest firing population of LC neurons during the active period. We also found that DMH orexin neurons project preferentially to LC and express a diurnal rhythm of activation that correlates with LC neuronal firing frequency. Therefore, we propose that DMH orexin neurons play a role in modulating the day–night differences of LC impulse activity. [Copyright &y& Elsevier]

Abstract: Corticotropin releasing hormone (CRH) plays a major role in central nervous system responses to stressors and has been implicated in stress-induced alterations in sleep. In the absence of stressors, CRH contributes to the regulation of spontaneous waking. We examined the effects of CRH and astressin (AST), a non-specific CRH antagonist, on wakefulness and sleep in two mouse strains with differential responsiveness to stress to determine whether CRH might also differentially affect undisturbed sleep and activity. Less reactive C57BL/6J (n =7) and high reactive BALB/cJ (n =7) male mice were implanted with a transmitter for determining sleep via telemetry and with a guide cannula aimed into a lateral ventricle. After recovery from surgery and habituation to handling, ICV microinjections of CRH (0.04, 0.2, and 0.4 μg), AST (0.1, 0.4, and 1.0 μg) or vehicle alone (pyrogen-free saline, 0.2 μl) were administered during the fourth hour after lights on and sleep was recorded for the subsequent 8 h. Comparisons of wakefulness and sleep were conducted across conditions and across strains. In C57BL/6J mice, REM was significantly decreased after microinjections of CRH (0.2 μg) and CRH (0.4 μg), and NREM and total sleep were decreased after microinjections of CRH (0.4 μg ). CRH (0.04 μg) and AST did not significantly change wakefulness or sleep. In BALB/cJ mice, CRH (0.4 μg) increased wakefulness and decreased NREM, REM and total sleep. AST decreased active wakefulness and significantly increased REM at the low and high dosages. These findings demonstrate that CRH produces changes in arousal when given to otherwise undisturbed mice. Strain differences in the effects of CRH and AST may be linked to the relative responsiveness of C57BL/6J and BALB/cJ mice to stressors and to underlying differences in the CRH system. [Copyright &y& Elsevier]

Abstract: Orexins A and B are a pair of neuropeptides implicated in the regulation of feeding and arousal behavior mediated through two orexin receptors type 1 and type 2. We have determined the arousal effects of newly developed selective orexin receptor type 2 agonist, [Ala11]orexin-B, on the sleep–wake cycle in rats. The effects of third ventricle intracerebroventricular (ICV) infusion of the novel orexin receptor type 2 selective agonist, [Ala11]orexin-B, on the sleep–wake cycle were investigated. ICV infusion of [Ala11]orexin-B (1, 10 and 40 nmol) during the light period (11:00–16:00) dose-dependently resulted in a significant increase in wake duration by 46.9% (n = 5, P &#60; 0.05), 159.2% (n = 4, P &#60; 0.01) and 163.6% (n = 7, P &#60; 0.01)), respectively, and a significant decrease in rapid eye movement (REM) (P &#60; 0.01) and non-REM sleep (P &#60; 0.01) for all doses. In contrast, ICV infusion of orexin B at the same doses (1, 10 and 40 nmol) caused a 16.6% (n = 6, non-significant), 99.8% (n = 6, P &#60; 0.05) and 72.0% (n = 4, P &#60; 0.05) increase in wakefulness, respectively. Moreover, orexin-A, which exerts its effects through orexin receptor type 1 and orexin receptor type 2 with similar potency, resulted in a significant increase in wakefulness duration by 17.1% (n = 6, P &#60; 0.05), 184.0% (n = 6, P &#60; 0.01) and 228.6% (n = 6, P &#60; 0.01) at doses of 0.1, 1 and 10 nmol, respectively. Further, the enhancement of wakefulness was accompanied by a marked reduction in REM and non-REM sleep. These findings suggest that orexin receptor type 2 plays an important role in the modulation of sleep–wake state and behavioral responses. [Copyright &y& Elsevier]

The participation of histaminergic H1 and noradrenergic α1 receptors in orexin A-induced wakefulness was studied by examining the sleep–wakefulness cycle in rats. Intracerebroventricular infusion of orexin A (1 nmol) caused an increase in the wakefulness state, while non-rapid eye movement sleep (NREM sleep) and rapid eye movement sleep (REM sleep) states were decreased. Prazosin (150 nmol) showed no significant antagonistic effect on the orexin A-induced increase in the wakefulness state and decrease in NREM and REM sleep. On the contrary, pyrilamine (150 nmol) was effective in antagonizing orexin A-induced increase in wakefulness and decrease in NREM sleep. When prazosin (150 nmol) and pyrilamine (150 nmol) were simultaneously perfused into the lateral ventricle, an almost complete antagonistic effect was observed with the increase in the wakefulness state and decrease in NREM sleep. Orexin A (1 nmol) caused a significant decrease in the histamine contents of the cortex, hippocampus and hypothalamus, whereas noradrenaline contents were decreased only in the hypothalamus. From these results, we concluded that the arousal effect induced by orexin A occurs through histaminergic H1 and noradrenergic α1 receptors, although participation of the H1 receptor was more important than the α1 receptor. [Copyright &y& Elsevier]

The number of hypocretin-containing neurons is markedly decreased in most patients with the sleep disorder narcolepsy. It is presently not known why the loss of hypocretin neurons occurs in these patients. In the present study, we tested the role of inflammation in the degeneration of hypocretin neurons. The proinflammagen lipopolysaccharide (LPS) was infused chronically for 30 days (flow rate=0.22 μg/h) into the lateral hypothalamus in rats. Compared with chronic infusions of phosphate-buffered saline (PBS), LPS infusions produced a decline in the number of hypocretin (29.7% reduction), melanin concentrating hormone (MCH; 24.7% reduction), and neuronal nuclear antigen (NeuN)-immunoreactive neurons, as well as a dense distribution of reactive astrocytes and microglia within the lateral hypothalamus. LPS infusions also produced a large increase in the amounts of wakefulness 6 days after the onset of infusion (72.5±8.7% of wakefulness during lights-on period compared with 45.3±1.8% in PBS-treated rats). Amounts of wakefulness returned to control levels in all LPS-treated rats 30 days after the onset of infusion. A single injection of LPS (1, 5, or 10 μg) did not produce a significant decline in the number of hypocretin, MCH, or NeuN-positive neurons. The loss of hypocretin neurons produced by chronic LPS administration suggests that inflammation may play a role in the loss of hypocretin neurons in narcolepsy. [Copyright &y& Elsevier]

The distribution of hypocretin-1 (hcrt-1) and hypocretin-2 (hcrt-2) immunoreactivities in the cat brainstem was examined using immunohistochemical techniques. Hcrt-1- and hcrt-2-positive fibers with varicosities were detected in almost all brainstem regions. However, no hcrt-1- or hcrt-2-immunoreactive neuronal somata were observed in the cat brainstem. Both hcrt-1- and hcrt-2-labeled fibers exhibited different densities in distinct regions of the brainstem. In most brainstem regions, the intensity of hcrt-1 immunoreactivity was higher than that of hcrt-2 immunoreactivity. The highest densities of hcrt-1- and hcrt-2-positive fibers were found in the nucleus raphe dorsalis (RD), the laterodorsal tegmental nucleus (LDT) and the locus coeruleus (LC), suggesting an important role for these peptides in functions related to sleep–wake behavior. [Copyright &y& Elsevier]

Histaminergic neurons of the tuberomammillary nucleus (TMN) are hypothesized to promote wakefulness, but little is known about the activity of these cells during spontaneous behavior. We measured histaminergic neuron activity in the dorsomedial, ventrolateral, and caudal TMN at four different times using Fos and adenosine deaminase immunohistochemistry and recordings of sleep/wake behavior. Because circadian factors could influence neuronal activity, we then assessed TMN neuron activity in predominantly sleeping or awake animals, all killed at the same time of day. In both experiments, Fos expression in histaminergic neurons of all three TMN subnuclei was higher during periods of wakefulness. These results demonstrate that histaminergic neurons throughout the TMN are wake-active, and this activity is largely independent of the time of day. [Copyright &y& Elsevier]

Orexin (ORX) A and B (hypocretins) are excitatory neuropeptides produced by neurons of the lateral hypothalamus that have been implicated in the regulation of the sleep–wake cycle. In rats, Fos (the product of the cfos gene) expression shows daily rhythms in areas involved in sleep and wakefulness and orexinergic neurons show elevated Fos expression during the night. The present study directly compared the daily pattern of Fos expression in orexinergic neurons in diurnal (A. niloticus; grass rats) and nocturnal (R. norvegicus; lab rats) rodents. Animals kept on a 12:12 light–dark cycle were perfused at six different Zeitgeber times (ZT), with lights on at ZT 0: 1, 5, 13, 17, 20 and 23. In both nocturnal and diurnal rodents orexinergic neurons showed rhythms in Fos expression, with more Fos seen during the active phase of each species. In the diurnal species, Fos expression in cells of the lateral hypothalamus that do not produce ORX was elevated at ZT 20, a time when these animals sleep, and was low at ZT 13, a time of peak of activity. These results provide further evidence for a link between activity in orexinergic neurons and wakefulness and that in grass rats, other neurons of the lateral hypothalamus may work in an antagonistic fashion with respect to orexinergic neurons to regulate wakefulness in this diurnal species. [Copyright &y& Elsevier]

The pedunculopontine tegmental nucleus (PPT) has been implicated in the generation and/or maintenance of both active sleep (AS) and wakefulness (W). GABAergic neurons are present within this nucleus and recent studies have shown that these neurons are active during AS . In order to examine the role of mesopontine GABAergic processes in the generation of AS, the GABAA agonist muscimol and the GABAA antagonist bicuculline were microinjected into the PPT of chronic cats that were prepared for recording the states of sleep and wakefulness. Muscimol increased the time spent in AS by increasing the frequency and duration of AS episodes; this increase in AS was at the expense of the time spent in wakefulness. A decrease in PGO density during AS was also observed following the microinjection of muscimol. On the other hand, bicuculline decreased both AS and quiet sleep and increased the time spent in wakefulness. These data suggest that GABA acts on GABAA receptors within the PPT to facilitate the generation of AS by suppressing the activity of waking-related processes within this nucleus. [Copyright &y& Elsevier]

Adenosine is a putative sleep factor with effects mainly mediated by the A1 receptor. Recent studies have implicated the hypothalamic orexin/hypocretin-containing neurons in the control of sleep–wakefulness. To help determine if adenosine might play a role in the control of orexin neurons, immunohistochemistry was used to characterize the distribution of adenosine A1 receptor protein on the orexinergic neurons. About 30% of orexin-containing neurons were labeled. The data supports the presence of adenosine A1 receptors on orexinergic neurons and suggests a possible substrate for a functional role of adenosine in the regulation of orexinergic activity. [Copyright &y& Elsevier]

Previous studies have shown that GABAergic processes in the ponto-mesencephalic region of the brainstem are involved in the generation of wakefulness and active sleep (AS). The dorsal and ventral tegmental nuclei of Gudden (DTN and VTN, respectively) are known to contain a large population of GABAergic neurons. In the present study, utilizing Fos immunoreactivity as a marker of neuronal activity, it was determined that GABAergic DTN pars dorsalis neurons are active during active wakefulness and AS induced by carbachol, but not during quiet wakefulness or quiet sleep. In contrast, no differences in the number of Fos immunoreactive neurons were observed in the DTN pars ventralis and VTN across behavioral states. [Copyright &y& Elsevier]

Terminals of hypothalamic hypocretin-containing neurons are observed within brainstem nuclei involved in the control of sleep and wakefulness. Because aged humans, cats and other species exhibit changes in sleep and wakefulness in old age, we were interested in examining age-related changes in hypocretin/orexin projections to the following brainstem regions which are associated with the regulation of sleep and wakefulness: the dorsal raphe nucleus, the laterodorsal tegmental nucleus, the pedunculo-pontine tegmental nucleus and the locus coeruleus. Based upon the results of immunohistochemical determinations, in all the regions examined, round or oval ‘spot-like’ structures were observed in aged cats. Many of these ‘spot-like’ structures resembled enlarged varicosities of a nature that would be expected to disrupt hypocretin neurotransmission. In addition, a site-specific decrease in immunostaining was observed in the locus coeruleus in old cats compared with adult controls; this result likely reflects a decrease in the number of labeled fibers, which indicates that there occurs a degeneration of hypocretinergic function in conjunction with old age. The proceeding changes may account for some of sleep–wake disturbance which are observed in aged animals as well as elderly humans. [Copyright &y& Elsevier]

The hippocampal theta rhythm (θ) was reported to be associated with movements, attention, auditory processing, autonomic functions, learning and memory and postulated as an associator of discontiguous events. Since visual information includes temporal cues, our study was centered on the correlation between hippocampal θ rhythm and lateral geniculate activity. Phase relationships between hippocampal θ and unit firing were found with both spontaneous and light evoked activity during wakefulness, slow wave and paradoxical sleep. This temporal correlation was dynamic, exhibiting changes related to the sleep–waking cycle and perhaps to attention shifts. Hippocampal θ rhythm may supply a low frequency temporal dimension to the processing of visual information. [Copyright &y& Elsevier]

Cats with chronically implanted electrodes were used to study changes in the auditory pathway during the wake/sleep cycle. Evoked responses to electrical stimulation of brain stem and thalamic nuclei were recorded and the animals' state of arousal was determined by observation and by monitoring EEG and EMG. Evoked responses in superior olive and inferior colliculus were invariant during the wake/sleep cycle. Evoked responses in medial geniculate and auditory radiations showed significant changes in slow wave sleep compared with wakefulness when stimulated from nuclei below inferior colliculus. However, with inferior colliculus stimulation, no significant changes in response at medial geniculate or radiations were observed. Evoked responses recorded from posterior A1 cortical sites showed increased amplitude of the late wave and simplification of the early biphasic waves during slow wave sleep. In contrast, evoked responses recorded from Ep and anterior A1 sites decreased in amplitude or were absent during slow wave sleep compared to responses recorded during wakefulness. No significant differences between evoked responses recorded at cortex during wake or REM sleep were obtained. It is suggested that in the auditory system, the major changes occurring during the wake/sleep cycle are cortical in origin, and sub-areas of auditory cortex change differentially. Some change is also observed in medial geniculate only if stimulation is below the level of inferior colliculus. © 1971.

Cats with chronically implanted electrodes were used to study changes in the auditory pathway during the wake/sleep cycle. Evoked responses to electrical stimulation of brain stem and thalamic nuclei were recorded and the animals' state of arousal was determined by observation and by monitoring EEG and EMG. Evoked responses in superior olive and inferior colliculus were invariant during the wake/sleep cycle. Evoked responses in medial geniculate and auditory radiations showed significant changes in slow wave sleep compared with wakefulness when stimulated from nuclei below inferior colliculus. However, with inferior colliculus stimulation, no significant changes in response at medial geniculate or radiations were observed. Evoked responses recorded from posterior A1 cortical sites showed increased amplitude of the late wave and simplification of the early biphasic waves during slow wave sleep. In contrast, evoked responses recorded from Ep and anterior A1 sites decreased in amplitude or were absent during slow wave sleep compared to responses recorded during wakefulness. No significant differences between evoked responses recorded at cortex during wake or REM sleep were obtained. It is suggested that in the auditory system, the major changes occurring during the wake/sleep cycle are cortical in origin, and sub-areas of auditory cortex change differentially. Some change is also observed in medial geniculate only if stimulation is below the level of inferior colliculus. © 1971.