Alpha anteriorization and theta posteriorization during deep sleep.

Brain states (wake, sleep, general anesthesia, etc.) are profoundly associated with the spatiotemporal dynamics of brain oscillations. Previous studies showed that the EEG alpha power shifted from the occipital cortex to the frontal cortex (alpha anteriorization) after being induced into a state of general anesthesia via propofol. The sleep research literature suggests that slow waves and sleep spindles are generated locally and propagated gradually to different brain regions. Since sleep and general anesthesia are conceptualized under the same framework of consciousness, the present study examines whether alpha anteriorization similarly occurs during sleep and how the EEG power in other frequency bands changes during different sleep stages. The results from the analysis of three polysomnography datasets of 234 participants show consistent alpha anteriorization during the sleep stages N2 and N3, beta anteriorization during stage REM, and theta posteriorization during stages N2 and N3. Although it is known that the neural circuits responsible for sleep are not exactly the same for general anesthesia, the findings of alpha anteriorization in this study suggest that, at macro level, the circuits for alpha oscillations are organized in the similar cortical areas. The spatial shifts of EEG power in different frequency bands during sleep may offer meaningful neurophysiological markers for the level of consciousness.

Sleepiness and the transition from wakefulness to sleep.

The transition from wakefulness to sleep is a progressive process that is reflected in the gradual loss of responsiveness, an alteration of cognitive functions, and a drastic shift in brain dynamics. These changes do not occur all at once. The sleep onset period (SOP) refers here to this period of transition between wakefulness and sleep. For example, although transitions of brain activity at sleep onset can occur within seconds in a given brain region, these changes occur at different time points across the brain, resulting in a SOP that can last several minutes. Likewise, the transition to sleep impacts cognitive and behavioral levels in a graded and staged fashion. It is often accompanied and preceded by a sensation of drowsiness and the subjective feeling of a need for sleep, also associated with specific physiological and behavioral signatures. To better characterize fluctuations in vigilance and the SOP, a multidimensional approach is thus warranted. Such a multidimensional approach could mitigate important limitations in the current classification of sleep, leading ultimately to better diagnoses and treatments of individuals with sleep and/or vigilance disorders. These insights could also be translated in real-life settings to either facilitate sleep onset in individuals with sleep difficulties or, on the contrary, prevent or control inappropriate sleep onsets.

Mechanisms of Activation of Brain's Drainage during Sleep: The Nightlife of Astrocytes.

The study of functions, mechanisms of generation, and pathways of movement of cerebral fluids has a long history, but the last decade has been especially productive. The proposed glymphatic hypothesis, which suggests a mechanism of the brain waste removal system (BWRS), caused an active discussion on both the criticism of some of the perspectives and our intensive study of new experimental facts. It was especially found that the intensity of the metabolite clearance changes significantly during the transition between sleep and wakefulness. Interestingly, at the cellular level, a number of aspects of this problem have been focused on, such as astrocytes-glial cells, which, over the past two decades, have been recognized as equal partners of neurons and perform many important functions. In particular, an important role was assigned to astrocytes within the framework of the glymphatic hypothesis. In this review, we return to the "astrocytocentric" view of the BWRS function and the explanation of its activation during sleep from the viewpoint of new findings over the last decade. Our main conclusion is that the BWRS's action may be analyzed both at the systemic (whole-brain) and at the local (cellular) level. The local level means here that the neuro-glial-vascular unit can also be regarded as the smallest functional unit of sleep, and therefore, the smallest functional unit of the BWRS.

What is sleep exactly? Global and local modulations of sleep oscillations all around the clock.

Wakefulness, non-rapid eye-movement (NREM) and rapid eye-movement (REM) sleep differ from each other along three dimensions: behavioral, phenomenological, physiological. Although these dimensions often fluctuate in step, they can also dissociate. The current paradigm that views sleep as made of global NREM and REM states fail to account for these dissociations. This conundrum can be dissolved by stressing the existence and significance of the local regulation of sleep. We will review the evidence in animals and humans, healthy and pathological brains, showing different forms of local sleep and the consequences on behavior, cognition, and subjective experience. Altogether, we argue that the notion of local sleep provides a unified account for a host of phenomena: dreaming in REM and NREM sleep, NREM and REM parasomnias, intrasleep responsiveness, inattention and mind wandering in wakefulness. Yet, the physiological origins of local sleep or its putative functions remain unclear. Exploring further local sleep could provide a unique and novel perspective on how and why we sleep.

Astrocytes: new evidence, new models, new roles.

Astrocytes have been in the limelight of active research for about 3 decades now. Over this period, ideas about their function and role in the nervous system have evolved from simple assistance in energy supply and homeostasis maintenance to a complex informational and metabolic hub that integrates data on local neuronal activity, sensory and arousal context, and orchestrates many crucial processes in the brain. Rapid progress in experimental techniques and data analysis produces a growing body of data, which can be used as a foundation for formulation of new hypotheses, building new refined mathematical models, and ultimately should lead to a new level of understanding of the contribution of astrocytes to the cognitive tasks performed by the brain. Here, we highlight recent progress in astrocyte research, which we believe expands our understanding of how low-level signaling at a cellular level builds up to processes at the level of the whole brain and animal behavior. We start our review with revisiting data on the role of noradrenaline-mediated astrocytic signaling in locomotion, arousal, sensory integration, memory, and sleep. We then briefly review astrocyte contribution to the regulation of cerebral blood flow regulation, which is followed by a discussion of biophysical mechanisms underlying astrocyte effects on different brain processes. The experimental section is closed by an overview of recent experimental techniques available for modulation and visualization of astrocyte dynamics. We then evaluate how the new data can be potentially incorporated into the new mathematical models or where and how it already has been done. Finally, we discuss an interesting prospect that astrocytes may be key players in important processes such as the switching between sleep and wakefulness and the removal of toxic metabolites from the brain milieu.

Tripping on the edge of consciousness.

Herein the major accomplishments, trials and tribulations, and epiphanies experienced by James M. Krueger over the course of his career in sleep research are presented. They include the characterization of a) the supranormal EEG delta waves occurring during NREMS post sleep loss, b) Factor S as a muramyl peptide, c) the physiological roles of cytokines in sleep regulation, d) multiple other sleep regulatory substances, e) the dramatic changes in sleep over the course of infectious diseases, and f) sleep initiation within small neuronal/glial networks. The theory that the preservation of brain plasticity is the primordial sleep function is briefly discussed. These accomplishments resulted from collaborations with many outstanding scientists including James M. Krueger's mentors (John Pappenheimer and Manfred Karnovsky) and collaborators later in life, including Charles Dinarello, Louis Chedid, Mark Opp, Ferenc Obal jr., Dave Rector, Ping Taishi, Linda Toth, Jeannine Majde, Levente Kapas, Eva Szentirmai, Jidong Fang, Chris Davis, Sandip Roy, Tetsuya Kushikata, Fabio Garcia-Garcia, Ilia Karatsoreos, Mark Zielinski, and Alok De, plus many students, e.g. Jeremy Alt, Kathryn Jewett, Erika English, and Victor Leyva-Grado.

Regional variability in intracerebral properties of NREM to REM sleep transitions in humans.

Transitions between wake and sleep states show a progressive pattern underpinned by local sleep regulation. In contrast, little evidence is available on non-rapid eye movement (NREM) to rapid eye movement (REM) sleep boundaries, considered as mainly reflecting subcortical regulation. Using polysomnography (PSG) combined with stereoelectroencephalography (SEEG) in humans undergoing epilepsy presurgical evaluation, we explored the dynamics of NREM-to-REM transitions. PSG was used to visually score transitions and identify REM sleep features. SEEG-based local transitions were determined automatically with a machine learning algorithm using features validated for automatic intra-cranial sleep scoring (10.5281/zenodo.7410501). We analyzed 2988 channel-transitions from 29 patients. The average transition time from all intracerebral channels to the first visually marked REM sleep epoch was 8 s ± 1 min 58 s, with a great heterogeneity between brain areas. Transitions were observed first in the lateral occipital cortex, preceding scalp transition by 1 min 57 s ± 2 min 14 s (d = -0.83), and close to the first sawtooth wave marker. Regions with late transitions were the inferior frontal and orbital gyri (1 min 1 s ± 2 min 1 s, d = 0.43, and 1 min 1 s ± 2 min 5 s, d = 0.43, after scalp transition). Intracranial transitions were earlier than scalp transitions as the night advanced (last sleep cycle, d = -0.81). We show a reproducible gradual pattern of REM sleep initiation, suggesting the involvement of cortical mechanisms of regulation. This provides clues for understanding oneiric experiences occurring at the NREM/REM boundary.

Sleep and vigilance states: Embracing spatiotemporal dynamics.

The classic view of sleep and vigilance states is a global stationary perspective driven by the interaction between neuromodulators and thalamocortical systems. However, recent data are challenging this view by demonstrating that vigilance states are highly dynamic and regionally complex. Spatially, sleep- and wake-like states often co-occur across distinct brain regions, as in unihemispheric sleep, local sleep in wakefulness, and during development. Temporally, dynamic switching prevails around state transitions, during extended wakefulness, and in fragmented sleep. This knowledge, together with methods monitoring brain activity across multiple regions simultaneously at millisecond resolution with cell-type specificity, is rapidly shifting how we consider vigilance states. A new perspective incorporating multiple spatial and temporal scales may have important implications for considering the governing neuromodulatory mechanisms, the functional roles of vigilance states, and their behavioral manifestations. A modular and dynamic view highlights novel avenues for finer spatiotemporal interventions to improve sleep function.

The Theta Paradox: 4-8 Hz EEG Oscillations Reflect Both Sleep Pressure and Cognitive Control.

Human electroencephalographic (EEG) oscillations characterize specific behavioral and vigilance states. The frequency of these oscillations is typically sufficient to distinguish a given state; however, theta oscillations (4-8 Hz) have instead been found in near-opposite conditions of drowsiness during sleep deprivation and alert cognitive control. While the latter has been extensively studied and is often referred to as "frontal midline theta," (fmTheta) the former has been investigated far less but is considered a marker for sleep pressure during wake. In this study we investigated to what extent theta oscillations differed during cognitive tasks and sleep deprivation. We measured high-density EEG in 18 young healthy adults (nine female) performing six tasks under three levels of sleep deprivation. We found both cognitive load and sleep deprivation increased theta power in medial prefrontal cortical areas; however, sleep deprivation caused additional increases in theta in many other, predominantly frontal, areas. The sources of sleep deprivation theta (sdTheta) were task dependent, with a visual-spatial task and short-term memory (STM) task showing the most widespread effects. Notably, theta was highest in supplementary motor areas during passive music listening, and highest in the inferior temporal cortex (responsible for object recognition) during a spatial game. Furthermore, while changes in task performance were correlated with increases in theta during sleep deprivation, this relationship was not specific to the EEG of the same task and did not survive correction for multiple comparisons. Altogether, these results suggest that both during sleep deprivation and cognition theta oscillations may preferentially occur in cortical areas not involved in ongoing behavior.SIGNIFICANCE STATEMENT Electroencephalographic (EEG) research in sleep has often remained separate from research in cognition. This has led to two incompatible interpretations of the function of theta brain oscillations (4-8 Hz): that they reflect local sleep events during sleep deprivation, or that they reflect cognitive processing during tasks. With this study, we found no fundamental differences between theta oscillations during cognition and theta during sleep deprivation that would suggest different functions. Instead, our results indicate that in both cases, theta oscillations are generated by cortical areas not required for ongoing behavior. Therefore, at least in humans, theta may reflect either cortical disengagement or inhibition.

Closed-loop modulation of local slow oscillations in human NREM sleep.

Slow-wave sleep is the deep non-rapid eye-movement (NREM) sleep stage that is most relevant for the recuperative function of sleep. Its defining property is the presence of slow oscillations (<2 Hz) in the scalp electroencephalogram (EEG). Slow oscillations are generated by a synchronous back and forth between highly active UP-states and silent DOWN-states in neocortical neurons. Growing evidence suggests that closed-loop sensory stimulation targeted at UP-states of EEG-defined slow oscillations can enhance the slow oscillatory activity, increase sleep depth, and boost sleep's recuperative functions. However, several studies failed to replicate such findings. Failed replications might be due to the use of conventional closed-loop stimulation algorithms that analyze the signal from one single electrode and thereby neglect the fact that slow oscillations vary with respect to their origins, distributions, and trajectories on the scalp. In particular, conventional algorithms nonspecifically target functionally heterogeneous UP-states of distinct origins. After all, slow oscillations at distinct sites of the scalp have been associated with distinct functions. Here we present a novel EEG-based closed-loop stimulation algorithm that allows targeting UP- and DOWN-states of distinct cerebral origins based on topographic analyses of the EEG: the topographic targeting of slow oscillations (TOPOSO) algorithm. We present evidence that the TOPOSO algorithm can detect and target local slow oscillations with specific, predefined voltage maps on the scalp in real-time. When compared to a more conventional, single-channel-based approach, TOPOSO leads to fewer but locally more specific stimulations in a simulation study. In a validation study with napping participants, TOPOSO targets auditory stimulation reliably at local UP-states over frontal, sensorimotor, and centro-parietal regions. Importantly, auditory stimulation temporarily enhanced the targeted local state. However, stimulation then elicited a standard frontal slow oscillation rather than local slow oscillations. The TOPOSO algorithm is suitable for the modulation and the study of the functions of local slow oscillations.

Electrophysiological Evidence of Local Sleep During Yoga Nidra Practice.

Background and Objectives: Yoga nidra is a technique sages use to self-induce sleep. Classically, sleep is characterized by three cardinal electrophysiological features, namely, electroencephalography (EEG), electromyography (EMG), and electrooculography (EOG). As the literature on electrophysiological characterization of Yoga nidra is lacking, it is not known whether it is a sleep or awake state. The objective of the study was to electrophysiologically characterize yoga nidra practice. Materials and Methods: Thirty subjects underwent five initial supervised yoga nidra sessions and then continued practice on their own. The subjects completed their sleep diaries for 2 weeks before and during the intervention. The electrophysiological characterization was done after 2 weeks of yoga nidra practice using 19 EEG channels polysomnography for pre-yoga nidra, yoga nidra practice and post-yoga nidra. Polysomnographic data were scored for sleep-wake stages as per standard criteria. Power spectral density (PSD) was calculated from various frequency bands in different time bins. EEG data were grouped by areas, namely, central, frontal, prefrontal, parietal, temporal, and occipital in time bins. Sleep diary parameters were also compared for pre-post-yoga nidra training. Results: After 2 weeks of yoga nidra practice, awake was scored throughout the session (n = 26). PSD results (mean difference in dB between different time bins; P value) showed significant changes. When compared to pre-yoga nidra, there was an increase in delta power in the central area (1.953; P = 0.033) and a decrease in the prefrontal area (2.713; P = 0.041) during yoga nidra. Sleep diary showed improvement in sleep duration (P = 0.0001), efficiency (P = 0.0005), quality (P = 0.0005), and total wake duration (P = 0.00005) after 2 weeks of practice. Interpretations and Conclusions: Yoga nidra practice in novices is electrophysiologically an awake state with signs of slow waves locally, often referred to as local sleep. Clinical Trial: Clinical Trial Registry of India, http://www.ctri.nic.in/Clinicaltrials/pmaindet2.php? trialid = 6253, 2013/05/003682.

SleepSEEG: automatic sleep scoring using intracranial EEG recordings only.

Objective.To perform automatic sleep scoring based only on intracranial electroencephalography (iEEG), without the need for scalp EEG), electrooculography (EOG) and electromyography (EMG), in order to study sleep, epilepsy, and their interaction.Approach. Data from 33 adult patients was used for development and training of the automatic scoring algorithm using both oscillatory and non-oscillatory spectral features. The first step consisted in unsupervised clustering of channels based on feature variability. For each cluster the classification was done in two steps, a multiclass tree followed by binary classification trees to distinguish the more challenging stage N1. The test data consisted in 11 patients, in whom the classification was done independently for each channel and then combined to get a single stage per epoch.Main results. An overall agreement of 78% was observed in the test set between the sleep scoring of the algorithm using iEEG alone and two human experts scoring based on scalp EEG, EOG and EMG. Balanced sensitivity and specificity were obtained for the different sleep stages. The performance was excellent for stages W, N2, and N3, and good for stage R, but with high variability across patients. The performance for the challenging stage N1 was poor, but at a similar level as for published algorithms based on scalp EEG. High confidence epochs in different stages (other than N1) can be identified with median per patient specificity >80%.Significance. The automatic algorithm can perform sleep scoring of long-term recordings of patients with intracranial electrodes undergoing presurgical evaluation in the absence of scalp EEG, EOG and EMG, which are normally required to define sleep stages but are difficult to use in the context of intracerebral studies. It also constitutes a valuable tool to generate hypotheses regarding local aspects of sleep, and will be significant for sleep evaluation in clinical epileptology and neuroscience research.

Cross-Scale Dynamicity of Entropy and Connectivity in the Sleeping Brain.

Introduction: The concept of local sleep refers to the phenomenon of local brain activity that modifies neural networks during unresponsive global sleep. Such network rewiring may differ across spatial scales; however, the global and local alterations in brain systems remain elusive in human sleep. Materials and Methods: We examined cross-scale changes of brain networks in sleep. Functional magnetic resonance imaging data were acquired from 28 healthy participants during nocturnal sleep. We adopted both metrics of connectivity (functional connectivity [FC] and regional homogeneity [ReHo]) and complexity (multiscale entropy) to explore the global and local functionality of the neural assembly across nonrapid eye movement sleep stages. Results: Long-range FC decreased with sleep depth, whereas local ReHo peaked at the N2 stage and reached its lowest level at the N3 stage. Entropy exhibited a general decline at the local scale (Scale 1) as sleep deepened, whereas the coarse-scale entropy (Scale 3) was consistent across stages. Discussion: The negative correlation between Scale-1 entropy and ReHo reflects the enhanced signal regularity and synchronization in sleep, identifying the information exchange at the local scale. The N2 stage showed a distinctive pattern toward local information processing with scrambled long-distance information exchange, indicating a specific time window for network reorganization. Collectively, the multidimensional metrics indicated an imbalanced global-local relationship among brain functional networks across sleep-wake stages.

Automated sleep state classification of wide-field calcium imaging data via multiplex visibility graphs and deep learning.

BACKGROUND: Wide-field calcium imaging (WFCI) allows for monitoring of cortex-wide neural dynamics in mice. When applied to the study of sleep, WFCI data are manually scored into the sleep states of wakefulness, non-REM (NREM) and REM by use of adjunct EEG and EMG recordings. However, this process is time-consuming and often suffers from low inter- and intra-rater reliability and invasiveness. Therefore, an automated sleep state classification method that operates on WFCI data alone is needed. NEW METHOD: A hybrid, two-step method is proposed. In the first step, spatial-temporal WFCI data is mapped to multiplex visibility graphs (MVGs). Subsequently, a two-dimensional convolutional neural network (2D CNN) is employed on the MVGs to be classified as wakefulness, NREM and REM. RESULTS: Sleep states were classified with an accuracy of 84% and Cohen's κ of 0.67. The method was also effectively applied on a binary classification of wakefulness/sleep (accuracy=0.82, κ = 0.62) and a four-class wakefulness/sleep/anesthesia/movement classification (accuracy=0.74, κ = 0.66). Gradient-weighted class activation maps revealed that the CNN focused on short- and long-term temporal connections of MVGs in a sleep state-specific manner. Sleep state classification performance when using individual brain regions was highest for the posterior area of the cortex and when cortex-wide activity was considered. COMPARISON WITH EXISTING METHOD: On a 3-hour WFCI recording, the MVG-CNN achieved a κ of 0.65, comparable to a κ of 0.60 corresponding to the human EEG/EMG-based scoring. CONCLUSIONS: The hybrid MVG-CNN method accurately classifies sleep states from WFCI data and will enable future sleep-focused studies with WFCI.

RoWDI: rolling window detection of sleep intrusions in the awake brain using fMRI.

Objective.Brief episodes of sleep can intrude into the awake human brain due to lack of sleep or fatigue-compromising the safety of critical daily tasks (i.e. driving). These intrusions can also introduce artefactual activity within functional magnetic resonance imaging (fMRI) experiments, prompting the need for an objective and effective method of removing them.Approach.We have developed a method to track sleep-like events in awake humans via rolling window detection of intrusions (RoWDI) of fMRI signal template. These events can then be used in voxel-wise event-related analysis of fMRI data. To test this approach, we generated a template of fMRI activity associated with transition to sleep via simultaneous fMRI and electroencephalogram (EEG) (N= 10). RoWDI was then used to identify sleep-like events in 20 individuals performing a cognitive task during fMRI after a night of partial sleep deprivation. This approach was further validated in an independent fMRI dataset (N= 56).Main results.Our method (RoWDI) was able to infer frequent sleep-like events during the cognitive task performed after sleep deprivation. The sleep-like events were associated with on average of 20% reduction in pupil size and prolonged response time. The blood-oxygen-level-dependent activity during the sleep-like events covered thalami-cortical regions, which although spatially distinct, co-existed with, task-related activity. These key findings were validated in the independent dataset.Significance.RoWDI can reliably detect spontaneous sleep-like events in the human brain. Thus, it may also be used as a tool to delineate and account for neural activity associated with wake-sleep transitions in both resting-state and task-related fMRI studies.

Mind-wandering Is Accompanied by Both Local Sleep and Enhanced Processes of Spatial Attention Allocation.

Mind-wandering (MW) is a subjective, cognitive phenomenon, in which thoughts move away from the task toward an internal train of thoughts, possibly during phases of neuronal sleep-like activity (local sleep, LS). MW decreases cortical processing of external stimuli and is assumed to decouple attention from the external world. Here, we directly tested how indicators of LS, cortical processing, and attentional selection change in a pop-out visual search task during phases of MW. Participants' brain activity was recorded using magnetoencephalography, MW was assessed via self-report using randomly interspersed probes. As expected, the performance decreased under MW. Consistent with the occurrence of LS, MW was accompanied by a decrease in high-frequency activity (HFA, 80-150 Hz) and an increase in slow wave activity (SWA, 1-6 Hz). In contrast, visual attentional selection as indexed by the N2pc component was enhanced during MW with the N2pc amplitude being directly linked to participants' performance. This observation clearly contradicts accounts of attentional decoupling that would predict a decrease in attention-related responses to external stimuli during MW. Together, our results suggest that MW occurs during phases of LS with processes of attentional target selection being upregulated, potentially to compensate for the mental distraction during MW.

Sleep-Wake Neurobiology.

Sleep and wakefulness are complex, tightly regulated behaviors that occur in virtually all animals. With recent exciting developments in neuroscience methodologies such as optogenetics, chemogenetics, and cell-specific calcium imaging technology, researchers can advance our understanding of how discrete neuronal groups precisely modulate states of sleep and wakefulness. In this chapter, we provide an overview of key neurotransmitter systems, neurons, and circuits that regulate states of sleep and wakefulness. We also describe long-standing models for the regulation of sleep/wake and non-rapid eye movement/rapid eye movement cycling. We contrast previous knowledge derived from classic approaches such as brain stimulation, lesions, cFos expression, and single-unit recordings, with emerging data using the newest technologies. Our understanding of neural circuits underlying the regulation of sleep and wakefulness is rapidly evolving, and this knowledge is critical for our field to elucidate the enigmatic function(s) of sleep.

The distinctive sleep pattern of the human calcarine cortex: a stereo-electroencephalographic study.

STUDY OBJECTIVES: The aim of this study was to describe the spontaneous electroencephalographic (EEG) features of sleep in the human calcarine cortex, comparing them with the well-established pattern of the parietal cortex. METHODS: We analyzed presurgical intracerebral EEG activity in calcarine and parietal cortices during non-rapid eye movement (NREM) and rapid eye movement (REM) sleep in seven patients with drug-resistant focal epilepsy. The time course of the EEG spectral power and NREM vs REM differences was assessed. Sleep spindles were automatically detected. To assess homeostatic dynamics, we considered the first vs second half of the night ratio in the delta frequency range (0.5-4 Hz) and the rise rate of delta activity during the first sleep cycle. RESULTS: While the parietal area showed the classically described NREM and REM sleep hallmarks, the calcarine cortex exhibited a distinctive pattern characterized by: (1) the absence of sleep spindles; (2) a large similarity between EEG power spectra of NREM and REM; and (3) reduced signs of homeostatic dynamics, with a decreased delta ratio between the first and the second half of the night, a reduced rise rate of delta activity during the first NREM sleep cycle, and lack of correlation between these measures. CONCLUSIONS: Besides describing for the first time the peculiar sleep EEG pattern in the human calcarine cortex, our findings provide evidence that different cortical areas may exhibit specific sleep EEG pattern, supporting the view of sleep as a local process and promoting the idea that the functional role of sleep EEG features should be considered at a regional level.

A wake-like state in vitro induced by transmembrane TNF/soluble TNF receptor reverse signaling.

Tumor necrosis factor alpha (TNF) has sleep regulatory and brain development roles. TNF promotes sleep in vivo and in vitro while TNF inhibition diminishes sleep. Transmembrane (tm) TNF and the tmTNF receptors (Rs), are cleaved by tumor necrosis factor alpha convertase to produce soluble (s) TNF and sTNFRs. Reverse signaling occurs in cells expressing tmTNF upon sTNFR binding. sTNFR administration in vivo inhibits sleep, thus we hypothesized that a wake-like state in vitro would be induced by sTNFR-tmTNF reverse signaling. Somatosensory cortical neuron/glia co-cultures derived from male and female mice lacking both TNFRs (TNFRKO), or lacking TNF (TNFKO) and wildtype (WT) mice were plated onto six-well multi-electrode arrays. Daily one-hour electrophysiological recordings were taken on culture days 4 through 14. sTNFR1 (0.0, 0.3, 3, 30, 60, and 120 ng/µL) was administered on day 14. A final one-hour recording was taken on day 15. Four measures were characterized that are also used to define sleep in vivo: action potentials (APs), burstiness index (BI), synchronization of electrical activity (SYN), and slow wave power (SWP; 0.25-3.75 Hz). Development rates of these emergent electrophysiological properties increased in cells from mice lacking TNF or both TNFRs compared to cells from WT mice. Decreased SWP, after the three lowest doses (0.3, 3 and 30 ng/µL) of the sTNFR1, indicate a wake-like state in cells from TNFRKO mice. A wake-like state was also induced after 3 ng/µl sTNFR1 treatment in cells from TNFKO mice, which express the TNFR1 ligand, lymphotoxin alpha. Cells from WT mice showed no treatment effects. Results are consistent with prior studies demonstrating involvement of TNF in brain development, TNF reverse signaling, and sleep regulation in vivo. Further, the current demonstration of sTNFR1 induction of a wake-like state in vitro is consistent with the idea that small neuronal/glial circuits manifest sleep- and wake-like states analogous to those occurring in vivo. Finally, that sTNF forward signaling enhances sleep while sTNFR1 reverse signaling enhances a wake-like state is consistent with other sTNF/tmTNF/sTNFR1 brain actions having opposing activities.

Diversity of simultaneous sleep in the motor cortex and hippocampus in rats.

We investigated the homogeneity/heterogeneity of spontaneous sleep, simultaneously recorded in the motor cortex and the hippocampus of control rats, and particularly analysed simultaneous and non-simultaneous motor cortical and hippocampal non-rapid eye movement (NREM)/rapid eye movement (REM) sleep. We demonstrate that the sleep architectures of the motor cortex and hippocampus are different in control rats. There was an increase of NREM duration and a decrease of REM duration in the hippocampus versus the motor cortex. In terms of duration, NREM state is the most heterogeneous in the hippocampus, whereas the REM state is the most heterogeneous in the motor cortex. Whereas the hippocampal NREM duration was increased due to the prolongation of NREM episodes, the hippocampal REM duration decreased due to the decreased number of REM episodes. The heterogeneity of sleep in the motor cortex and hippocampus in control rats was particularly expressed through the inverse alteration of sigma amplitude during NREM sleep and beta/gamma amplitudes during REM sleep in the hippocampus, along with the delta, sigma, beta and gamma amplitudes only during non-simultaneous NREM/REM sleep in the hippocampus. We demonstrated the brain structure-related and NREM/REM state-related heterogeneity of the motor cortical and hippocampal local sleep in control rats. The distinctly altered local NREM/REM states, alongside their episode dynamics and electroencephalographic (EEG) microstructures, suggest the importance of both the local neuronal network substrate and the NREM/REM neurochemical substrate in the control mechanisms of sleep.