Spinal projecting neurons in rostral ventromedial medulla co-regulate motor and sympathetic tone.

Many behaviors require the coordinated actions of somatic and autonomic functions. However, the underlying mechanisms remain elusive. By opto-stimulating different populations of descending spinal projecting neurons (SPNs) in anesthetized mice, we show that stimulation of excitatory SPNs in the rostral ventromedial medulla (rVMM) resulted in a simultaneous increase in somatomotor and sympathetic activities. Conversely, opto-stimulation of rVMM inhibitory SPNs decreased both activities. Anatomically, these SPNs innervate both sympathetic preganglionic neurons and motor-related regions in the spinal cord. Fiber-photometry recording indicated that the activities of rVMM SPNs correlate with different levels of muscle and sympathetic tone during distinct arousal states. Inhibiting rVMM excitatory SPNs reduced basal muscle and sympathetic tone, impairing locomotion initiation and high-speed performance. In contrast, silencing the inhibitory population abolished muscle atonia and sympathetic hypoactivity during rapid eye movement (REM) sleep. Together, these results identify rVMM SPNs as descending spinal projecting pathways controlling the tone of both the somatomotor and sympathetic systems.

Individualized temporal patterns dominate cortical upstate and sleep depth in driving human sleep spindle timing.

Sleep spindles are critical for memory consolidation and strongly linked to neurological disease and aging. Despite their significance, the relative influences of factors like sleep depth, cortical up/down states, and spindle temporal patterns on individual spindle production remain poorly understood. Moreover, spindle temporal patterns are typically ignored in favor of an average spindle rate. Here, we analyze spindle dynamics in 1008 participants from the Multi-Ethnic Study of Atherosclerosis using a point process framework. Results reveal fingerprint-like temporal patterns, characterized by a refractory period followed by a period of increased spindle activity, which are highly individualized yet consistent night-to-night. We observe increased timing variability with age and distinct gender/age differences. Strikingly, and in contrast to the prevailing notion, individualized spindle patterns are the dominant determinant of spindle timing, accounting for over 70% of the statistical deviance explained by all of the factors we assessed, surpassing the contribution of slow oscillation (SO) phase (~14%) and sleep depth (~16%). Furthermore, we show spindle/SO coupling dynamics with sleep depth are preserved across age, with a global negative shift towards the SO rising slope. These findings offer novel mechanistic insights into spindle dynamics with direct experimental implications and applications to individualized electroencephalography biomarker identification.

Transient oscillation dynamics during sleep provide a robust basis for electroencephalographic phenotyping and biomarker identification.

Transient oscillatory events in the sleep electroencephalogram represent short-term coordinated network activity. Of particular importance, sleep spindles are transient oscillatory events associated with memory consolidation, which are altered in aging and in several psychiatric and neurodegenerative disorders. Spindle identification, however, currently contains implicit assumptions derived from what waveforms were historically easiest to discern by eye, and has recently been shown to select only a high-amplitude subset of transient events. Moreover, spindle activity is typically averaged across a sleep stage, collapsing continuous dynamics into discrete states. What information can be gained by expanding our view of transient oscillatory events and their dynamics? In this paper, we develop a novel approach to electroencephalographic phenotyping, characterizing a generalized class of transient time-frequency events across a wide frequency range using continuous dynamics. We demonstrate that the complex temporal evolution of transient events during sleep is highly stereotyped when viewed as a function of slow oscillation power (an objective, continuous metric of depth-of-sleep) and phase (a correlate of cortical up/down states). This two-fold power-phase representation has large intersubject variability-even within healthy controls-yet strong night-to-night stability for individuals, suggesting a robust basis for phenotyping. As a clinical application, we then analyze patients with schizophrenia, confirming established spindle (12-15 Hz) deficits as well as identifying novel differences in transient non-rapid eye movement events in low-alpha (7-10 Hz) and theta (4-6 Hz) ranges. Overall, these results offer an expanded view of transient activity, describing a broad class of events with properties varying continuously across spatial, temporal, and phase-coupling dimensions.

Sources of Variation in the Spectral Slope of the Sleep EEG.

The 1/f spectral slope of the electroencephalogram (EEG) estimated in the γ frequency range has been proposed as an arousal marker that differentiates wake, nonrapid eye movement (NREM) sleep, and rapid eye movement (REM) sleep. Here, we sought to replicate and extend these findings in a large sample, providing a comprehensive characterization of how slope changes with age, sex, and its test-retest reliability as well as potential confounds that could affect the slope estimation. We used 10,255 whole-night polysomnograms (PSGs) from the National Sleep Research Resource (NSRR). All preprocessing steps were performed using an open-source Luna package and the spectral slope was estimated by fitting log-log linear regression models on the absolute power from 30 to 45 Hz separately for wake, NREM, and REM stages. We confirmed that the mean spectral slope grows steeper going from wake to NREM to REM sleep. We found that the choice of mastoid referencing scheme modulated the extent to which electromyogenic, or electrocardiographic artifacts were likely to bias 30- to 45-Hz slope estimates, as well as other sources of technical, device-specific bias. Nonetheless, within individuals, slope estimates were relatively stable over time. Both cross-sectionally and longitudinal, slopes tended to become shallower with increasing age, particularly for REM sleep; males tended to show flatter slopes than females across all states. Our findings support that spectral slope can be a valuable arousal marker for both clinical and research endeavors but also underscore the importance of considering interindividual variation and multiple methodological aspects related to its estimation.

Dynamic models of obstructive sleep apnea provide robust prediction of respiratory event timing and a statistical framework for phenotype exploration.

Obstructive sleep apnea (OSA), in which breathing is reduced or ceased during sleep, affects at least 10% of the population and is associated with numerous comorbidities. Current clinical diagnostic approaches characterize severity and treatment eligibility using the average respiratory event rate over total sleep time (apnea-hypopnea index). This approach, however, does not characterize the time-varying and dynamic properties of respiratory events that can change as a function of body position, sleep stage, and previous respiratory event activity. Here, we develop a statistical model framework based on point process theory that characterizes the relative influences of all these factors on the moment-to-moment rate of event occurrence. Our results provide new insights into the temporal dynamics of respiratory events, suggesting that most adults have a characteristic event pattern that involves a period of normal breathing followed by a period of increased probability of respiratory event occurrence, while significant differences in event patterns are observed among gender, age, and race/ethnicity groups. Statistical goodness-of-fit analysis suggests consistent and substantial improvements in our ability to capture the timing of individual respiratory events using our modeling framework. Overall, we demonstrate a more statistically robust approach to characterizing sleep disordered breathing that can also serve as a basis for identifying future patient-specific respiratory phenotypes, providing an improved pathway towards developing individualized treatments.

Sleep spindles comprise a subset of a broader class of electroencephalogram events.

STUDY OBJECTIVES: Sleep spindles are defined based on expert observations of waveform features in the electroencephalogram (EEG) traces. This is a potentially limiting characterization, as transient oscillatory bursts like spindles are easily obscured in the time domain by higher amplitude activity at other frequencies or by noise. It is therefore highly plausible that many relevant events are missed by current approaches based on traditionally defined spindles. Given their oscillatory structure, we reexamine spindle activity from first principles, using time-frequency activity in comparison to scored spindles. METHODS: Using multitaper spectral analysis, we observe clear time-frequency peaks in the sigma (10-16 Hz) range (TFσ peaks). While nearly every scored spindle coincides with a TFσ peak, numerous similar TFσ peaks remain undetected. We therefore perform statistical analyses of spindles and TFσ peaks using manual and automated detection methods, comparing event cooccurrence, morphological similarities, and night-to-night consistency across multiple datasets. RESULTS: On average, TFσ peaks have more than three times the rate of spindles (mean rate: 9.8 vs. 3.1 events/minute). Moreover, spindles subsample the most prominent TFσ peaks with otherwise identical spectral morphology. We further demonstrate that detected TFσ peaks have stronger night-to-night rate stability (ρ = 0.98) than spindles (ρ = 0.67), while covarying with spindle rates across subjects (ρ = 0.72). CONCLUSIONS: These results provide compelling evidence that traditionally defined spindles constitute a subset of a more generalized class of EEG events. TFσ peaks are therefore a more complete representation of the underlying phenomenon, providing a more consistent and robust basis for future experiments and analyses.

Macro and micro sleep architecture and cognitive performance in older adults.

We sought to determine which facets of sleep neurophysiology were most strongly linked to cognitive performance in 3,819 older adults from two independent cohorts, using whole-night electroencephalography. From over 150 objective sleep metrics, we identified 23 that predicted cognitive performance, and processing speed in particular, with effects that were broadly independent of gross changes in sleep quality and quantity. These metrics included rapid eye movement duration, features of the electroencephalography power spectra derived from multivariate analysis, and spindle and slow oscillation morphology and coupling. These metrics were further embedded within broader associative networks linking sleep with aging and cardiometabolic disease: individuals who, compared with similarly aged peers, had better cognitive performance tended to have profiles of sleep metrics more often seen in younger, healthier individuals. Taken together, our results point to multiple facets of sleep neurophysiology that track coherently with underlying, age-dependent determinants of cognitive and physical health trajectories in older adults.

Estimation of Time-Varying Spectral Peaks and Decomposition of EEG Spectrograms.

Detection of spectral peaks and estimation of their properties, including frequency and amplitude, are fundamental to many applications of signal processing. Electroencephalography (EEG) of sleep, in particular, displays characteristic oscillations that change continuously throughout the night. Capturing these dynamics is essential to understanding the sleep process and characterizing the heterogeneity observed across individuals. Most sleep EEG analyses rely on either time-averaged spectra or bandpassed amplitude/power. Unfortunately, these approaches obscure the time-variability of peak properties, require specification of a priori criteria, and cannot distinguish power from nearby oscillations. More sophisticated approaches, using various spectral models, have been proposed to better estimate oscillatory properties, but these too have limitations. We present an improved approach to spectrogram decomposition, tracking time-varying parameterized peak functions and dynamically estimating their parameters using a modified form of the iterated extended Kalman filter (IEKF) that incorporates discrete On/Off-switching of peak combinations and a sampling step to draw the initial reference trajectory. We evaluate this approach on two types of simulated examples-one nearly within the model class and one outside. We find excellent performance, in terms of spectral fits and accuracy of estimated states, for both simulation types. We then apply the approach to real EEG data of sleep onset, obtaining quality spectral estimates with estimated peak combinations closely matching the expert-scored sleep stages. This approach offers not only the ability to estimate time-varying parameters of spectral peaks but, moving forward, the potential to estimate the governing dynamics and analyze their variability across nights, subjects, and clinical groups.

Quantifying statistical uncertainty in metrics of sleep disordered breathing.

BACKGROUND: The apnea-hypopnea index (AHI) (or one of its derivatives) is the primary clinical metric for characterizing sleep disordered breathing-the value of which with respect to a threshold determines severity of diagnosis and eligibility for treatment reimbursement. The index value, however, is taken as a perfect point estimate, with no measure of statistical uncertainty. Thus, current practice does not robustly account for variability in diagnosis/eligibility due to chance. In this paper, we quantify the statistical uncertainty associated with respiratory event indices for sleep disordered breathing and the effect of uncertainty on treatment eligibility. METHODS: We develop an empirical estimate of uncertainty using a non-parametric bootstrap on the interevent times, as well as a theoretical Poisson estimate reflecting the current formulation of the AHI. We then apply these methods to estimate AHI uncertainty for 2049 subjects (954/1095 M/F, age: mean 69 ± 9.1) from the Multi-Ethnic Study of Atherosclerosis (MESA). RESULTS AND CONCLUSIONS: The mean 95% empirical confidence interval width was 11.500 ± 6.208 events per hour and the mean 95% theoretical Poisson confidence interval width was 5.998 ± 2.897 events per hour, suggesting that uncertainty is likely a major confounding factor within the current diagnostic framework. Of the 278 subjects in the symptomatic population (ESS>10), 27% (76/278) had uncertain diagnoses given the 95% empirical confidence interval. Of the 2049 subjects in the full population, 43% (880/2049) had uncertain diagnoses given the 95% empirical confidence interval. The inclusion of subjects with uncertain diagnoses increases the number of eligible patients by 21.3% for the symptomatic population and by 84.8% for the full population. The exclusion of subjects with uncertain diagnoses given the 95% empirical confidence interval decreases the number of eligible patients by 12.4% for the symptomatic population and by 34.8% for full population. Additional analyses suggest that it is practically infeasible to gain diagnostic statistical significance through additional testing for a broad range of borderline cases. Overall, these results suggest that AHI uncertainty is a vital additional piece of information that would greatly benefit clinical practice, and that the inclusion of uncertainty in epidemiological analysis might help improve the ability for researchers to robustly link AHI with co-morbidities and long-term outcomes.

Sparse Multi-task Inverse Covariance Estimation for Connectivity Analysis in EEG Source Space.

Understanding how different brain areas interact to generate complex behavior is a primary goal of neuroscience research. One approach, functional connectivity analysis, aims to characterize the connectivity patterns within brain networks. In this paper, we address the problem of discriminative connectivity, i.e. determining the differences in network structure under different experimental conditions. We introduce a novel model called Sparse Multi-task Inverse Covariance Estimation (SMICE) which is capable of estimating a common connectivity network as well as discriminative networks across different tasks. We apply the method to EEG signals after solving the inverse problem of source localization, yielding networks defined on the cortical surface. We propose an efficient algorithm based on the Alternating Direction Method of Multipliers (ADMM) to solve SMICE. We apply our newly developed framework to find common and discriminative connectivity patterns for α-oscillations during the Sleep Onset Process (SOP) and during Rapid Eye Movement (REM) sleep. Even though both stages exhibit a similar α-oscillations, we show that the underlying networks are distinct.

Multitaper Infinite Hidden Markov Model for EEG.

Electroencephalographam (EEG) monitoring of neural activity is widely used for identifying underlying brain states. For inference of brain states, researchers have often used Hidden Markov Models (HMM) with a fixed number of hidden states and an observation model linking the temporal dynamics embedded in EEG to the hidden states. The use of fixed states may be limiting, in that 1) pre-defined states might not capture the heterogeneous neural dynamics across individuals and 2) the oscillatory dynamics of the neural activity are not directly modeled. To this end, we use a Hierarchical Dirichlet Process Hidden Markov Model (HDP-HMM), which discovers the set of hidden states that best describes the EEG data, without a-priori specification of state number. In addition, we introduce an observation model based on classical asymptotic results of frequency domain properties of stationary time series, along with the description of the conditional distributions for Gibbs sampler inference. We then combine this with multitaper spectral estimation to reduce the variance of the spectral estimates. By applying our method to simulated data inspired by sleep EEG, we arrive at two main results: 1) the algorithm faithfully recovers the spectral characteristics of the true states, as well as the right number of states and 2) the incorporation of the multitaper framework produces a more stable estimate than traditional periodogram spectral estimates.

Sleep Neurophysiological Dynamics Through the Lens of Multitaper Spectral Analysis.

During sleep, cortical and subcortical structures within the brain engage in highly structured oscillatory dynamics that can be observed in the electroencephalogram (EEG). The ability to accurately describe changes in sleep state from these oscillations has thus been a major goal of sleep medicine. While numerous studies over the past 50 years have shown sleep to be a continuous, multifocal, dynamic process, long-standing clinical practice categorizes sleep EEG into discrete stages through visual inspection of 30-s epochs. By representing sleep as a coarsely discretized progression of stages, vital neurophysiological information on the dynamic interplay between sleep and arousal is lost. However, by using principled time-frequency spectral analysis methods, the rich dynamics of the sleep EEG are immediately visible-elegantly depicted and quantified at time scales ranging from a full night down to individual microevents. In this paper, we review the neurophysiology of sleep through this lens of dynamic spectral analysis. We begin by reviewing spectral estimation techniques traditionally used in sleep EEG analysis and introduce multitaper spectral analysis, a method that makes EEG spectral estimates clearer and more accurate than traditional approaches. Through the lens of the multitaper spectrogram, we review the oscillations and mechanisms underlying the traditional sleep stages. In doing so, we will demonstrate how multitaper spectral analysis makes the oscillatory structure of traditional sleep states instantaneously visible, closely paralleling the traditional hypnogram, but with a richness of information that suggests novel insights into the neural mechanisms of sleep, as well as novel clinical and research applications.

Tracking the sleep onset process: an empirical model of behavioral and physiological dynamics.

The sleep onset process (SOP) is a dynamic process correlated with a multitude of behavioral and physiological markers. A principled analysis of the SOP can serve as a foundation for answering questions of fundamental importance in basic neuroscience and sleep medicine. Unfortunately, current methods for analyzing the SOP fail to account for the overwhelming evidence that the wake/sleep transition is governed by continuous, dynamic physiological processes. Instead, current practices coarsely discretize sleep both in terms of state, where it is viewed as a binary (wake or sleep) process, and in time, where it is viewed as a single time point derived from subjectively scored stages in 30-second epochs, effectively eliminating SOP dynamics from the analysis. These methods also fail to integrate information from both behavioral and physiological data. It is thus imperative to resolve the mismatch between the physiological evidence and analysis methodologies. In this paper, we develop a statistically and physiologically principled dynamic framework and empirical SOP model, combining simultaneously-recorded physiological measurements with behavioral data from a novel breathing task requiring no arousing external sensory stimuli. We fit the model using data from healthy subjects, and estimate the instantaneous probability that a subject is awake during the SOP. The model successfully tracked physiological and behavioral dynamics for individual nights, and significantly outperformed the instantaneous transition models implicit in clinical definitions of sleep onset. Our framework also provides a principled means for cross-subject data alignment as a function of wake probability, allowing us to characterize and compare SOP dynamics across different populations. This analysis enabled us to quantitatively compare the EEG of subjects showing reduced alpha power with the remaining subjects at identical response probabilities. Thus, by incorporating both physiological and behavioral dynamics into our model framework, the dynamics of our analyses can finally match those observed during the SOP.

A comparison of propofol- and dexmedetomidine-induced electroencephalogram dynamics using spectral and coherence analysis.

BACKGROUND: Electroencephalogram patterns observed during sedation with dexmedetomidine appear similar to those observed during general anesthesia with propofol. This is evident with the occurrence of slow (0.1 to 1 Hz), delta (1 to 4 Hz), propofol-induced alpha (8 to 12 Hz), and dexmedetomidine-induced spindle (12 to 16 Hz) oscillations. However, these drugs have different molecular mechanisms and behavioral properties and are likely accompanied by distinguishing neural circuit dynamics. METHODS: The authors measured 64-channel electroencephalogram under dexmedetomidine (n = 9) and propofol (n = 8) in healthy volunteers, 18 to 36 yr of age. The authors administered dexmedetomidine with a 1-µg/kg loading bolus over 10 min, followed by a 0.7 µg kg h infusion. For propofol, the authors used a computer-controlled infusion to target the effect-site concentration gradually from 0 to 5 µg/ml. Volunteers listened to auditory stimuli and responded by button press to determine unconsciousness. The authors analyzed the electroencephalogram using multitaper spectral and coherence analysis. RESULTS: Dexmedetomidine was characterized by spindles with maximum power and coherence at approximately 13 Hz (mean ± SD; power, -10.8 ± 3.6 dB; coherence, 0.8 ± 0.08), whereas propofol was characterized with frontal alpha oscillations with peak frequency at approximately 11 Hz (power, 1.1 ± 4.5 dB; coherence, 0.9 ± 0.05). Notably, slow oscillation power during a general anesthetic state under propofol (power, 13.2 ± 2.4 dB) was much larger than during sedative states under both propofol (power, -2.5 ± 3.5 dB) and dexmedetomidine (power, -0.4 ± 3.1 dB). CONCLUSION: The results indicate that dexmedetomidine and propofol place patients into different brain states and suggest that propofol enables a deeper state of unconsciousness by inducing large-amplitude slow oscillations that produce prolonged states of neuronal silence.

Characterizing context-dependent differential firing activity in the hippocampus and entorhinal cortex.

The rat hippocampus and entorhinal cortex have been shown to possess neurons with place fields that modulate their firing properties under different behavioral contexts. Such context-dependent changes in neural activity are commonly studied through electrophysiological experiments in which a rat performs a continuous spatial alternation task on a T-maze. Previous research has analyzed context-based differential firing during this task by describing differences in the mean firing activity between left-turn and right-turn experimental trials. In this article, we develop qualitative and quantitative methods to characterize and compare changes in trial-to-trial firing rate variability for sets of experimental contexts. We apply these methods to cells in the CA1 region of hippocampus and in the dorsocaudal medial entorhinal cortex (dcMEC), characterizing the context-dependent differences in spiking activity during spatial alternation. We identify a subset of cells with context-dependent changes in firing rate variability. Additionally, we show that dcMEC populations encode turn direction uniformly throughout the T-maze stem, whereas CA1 populations encode context at major waypoints in the spatial trajectory. Our results suggest scenarios in which individual cells that sparsely provide information on turn direction might combine in the aggregate to produce a robust population encoding.

Tracking non-stationary spectral peak structure in EEG data.

We develop a particle filter algorithm to simultaneously estimate and track the instantaneous peak frequency, amplitude, and bandwidth of multiple concurrent non-stationary components of an EEG signal in the time-frequency domain. We use this method to characterize human EEG activity during anesthesia-induced unconsciousness.

Electroencephalogram signatures of loss and recovery of consciousness from propofol.

Unconsciousness is a fundamental component of general anesthesia (GA), but anesthesiologists have no reliable ways to be certain that a patient is unconscious. To develop EEG signatures that track loss and recovery of consciousness under GA, we recorded high-density EEGs in humans during gradual induction of and emergence from unconsciousness with propofol. The subjects executed an auditory task at 4-s intervals consisting of interleaved verbal and click stimuli to identify loss and recovery of consciousness. During induction, subjects lost responsiveness to the less salient clicks before losing responsiveness to the more salient verbal stimuli; during emergence they recovered responsiveness to the verbal stimuli before recovering responsiveness to the clicks. The median frequency and bandwidth of the frontal EEG power tracked the probability of response to the verbal stimuli during the transitions in consciousness. Loss of consciousness was marked simultaneously by an increase in low-frequency EEG power (<1 Hz), the loss of spatially coherent occipital alpha oscillations (8-12 Hz), and the appearance of spatially coherent frontal alpha oscillations. These dynamics reversed with recovery of consciousness. The low-frequency phase modulated alpha amplitude in two distinct patterns. During profound unconsciousness, alpha amplitudes were maximal at low-frequency peaks, whereas during the transition into and out of unconsciousness, alpha amplitudes were maximal at low-frequency nadirs. This latter phase-amplitude relationship predicted recovery of consciousness. Our results provide insights into the mechanisms of propofol-induced unconsciousness, establish EEG signatures of this brain state that track transitions in consciousness precisely, and suggest strategies for monitoring the brain activity of patients receiving GA.

A general likelihood framework for characterizing the time course of neural activity.

We develop a general likelihood-based framework for use in the estimation of neural firing rates, which is designed to choose the temporal smoothing parameters that maximize the likelihood of missing data. This general framework is algorithm-independent and thus can be applied to a multitude of established methods for firing rate or conditional intensity estimation. As a simple example of the use of the general framework, we apply it to the peristimulus time histogram and kernel smoother, the methods most widely used for firing rate estimation in the electrophysiological literature and practice. In doing so, we illustrate how the use of the framework can employ the general point process likelihood as a principled cost function and can provide substantial improvements in estimation accuracy for even the most basic of rate estimation algorithms. In particular, the resultant kernel smoother is simple to implement, efficient to compute, and can accurately determine the bandwidth of a given rate process from individual spike trains. We perform a simulation study to illustrate how the likelihood framework enables the kernel smoother to pick the bandwidth parameter that best predicts missing data, and we show applications to real experimental spike train data. Additionally, we discuss how the general likelihood framework may be used in conjunction with more sophisticated methods for firing rate and conditional intensity estimation and suggest possible applications.

Phase-based measures of cross-frequency coupling in brain electrical dynamics under general anesthesia.

The state of general anesthesia (GA) is associated with an increase in spectral power in scalp electroencephalogram (EEG) at frequencies below 40 Hz, including spectral peaks in the slow oscillation (SO, 0.1-1 Hz) and α (8-14 Hz) bands. Because conventional power spectral analyses are insensitive to possible cross-frequency coupling, the relationships among the oscillations at different frequencies remain largely unexplored. Quantifying such coupling is essential for improving clinical monitoring of anesthesia and understanding the neuroscience of this brain state. We tested the usefulness of two measures of cross-frequency coupling: the bispectrum-derived SynchFastSlow, which is sensitive to phase-phase coupling in different frequency bands, and modulogram analysis of coupling between SO phase and α rhythm amplitude. SynchFastSlow, a metric that is used in clinical depth-of-anesthesia monitors, showed a robust correlation with the loss of consciousness at the induction of propofol GA, but this could be largely explained by power spectral changes without considering cross-frequency coupling. Modulogram analysis revealed two distinct modes of cross-frequency coupling under GA. The waking and two distinct states under GA could be discriminated by projecting in a two-dimensional phase space defined by the SynchFastSlow and the preferred SO phase of α activity. Our results show that a stereotyped pattern of phase-amplitude coupling accompanies multiple stages of anesthetic-induced unconsciousness. These findings suggest that modulogram analysis can improve EEG based monitoring of brain state under GA.