Glial Gap Junction Pathology in the Spinal Cord of the 5xFAD Mouse Model of Early-Onset Alzheimer's Disease.

Gap junctions (GJs) are specialized transmembrane channels assembled by two hemi-channels of six connexin (Cx) proteins that facilitate neuroglial crosstalk in the central nervous system (CNS). Previous studies confirmed the crucial role of glial GJs in neurodegenerative disorders with dementia or motor dysfunction including Alzheimer's disease (AD). The aim of this study was to examine the alterations in astrocyte and related oligodendrocyte GJs in association with Aß plaques in the spinal cord of the 5xFAD mouse model of AD. Our analysis revealed abundant Aß plaque deposition, activated microglia, and astrogliosis in 12-month-old (12M) 5xFAD mice, with significant impairment of motor performance starting from 3-months (3M) of age. Additionally, 12M 5xFAD mice displayed increased immunoreactivity of astroglial Cx43 and Cx30 surrounding Aß plaques and higher protein levels, indicating upregulated astrocyte-to-astrocyte GJ connectivity. In addition, they demonstrated increased numbers of mature CC1-positive and precursor oligodendrocytes (OPCs) with higher immunoreactivity of Cx47-positive GJs in individual cells. Moreover, total Cx47 protein levels were significantly elevated in 12M 5xFAD, reflecting increased oligodendrocyte-to-oligodendrocyte Cx47-Cx47 GJ connectivity. In contrast, we observed a marked reduction in Cx32 protein levels in 12M 5xFAD spinal cords compared with controls, while qRT-PCR analysis revealed a significant upregulation in Cx32 mRNA levels. Finally, myelin deficits were found focally in the areas occupied by Aß plaques, whereas axons themselves remained preserved. Overall, our data provide novel insights into the altered glial GJ expression in the spinal cord of the 5xFAD model of AD and the implicated role of GJ pathology in neurodegeneration. Further investigation to understand the functional consequences of these extensive alterations in oligodendrocyte-astrocyte (O/A) GJ connectivity is warranted.

Spatiotemporal profiles of focal and generalised spikes in childhood absence epilepsy.

The EEG in childhood absence epilepsy (CAE) may contain focal and generalised spike-wave discharges (SWDs) with focal, mainly frontal, "lead-in". The term "frontal absence" has been used to imply fast, secondary, 3-Hz generalisation from occult frontal foci with potential impact on clinical EEG interpretation and syndrome classification. The aim of this study was to investigate the relationship between focal and generalised SWDs. We studied five children with CAE and examined a sufficient number of focal ("interictal") and generalised SWDs in order to obtain reliable analysis. All generalised SWDs with focal lead-in were "decomposed" into their "pre-generalisation" focal and "generalised" constituents, which were studied separately. Two types of focal SWD ("interictal" and "pre-generalisation") and generalised SWD were visually clustered into groups, waveform-averaged, and plotted in the 2D-electrode space. Spatiotemporal analysis demonstrated a variety (mean: 4.2 per child; SD: 2.12) of mainly frontal and occipital locations for pre-generalisation focal SWDs with propagation along the longitudinal axis in either direction and across homologous sites. Interictal focal SWDs demonstrated similar spatiotemporal characteristics. In contrast, the topography and propagation patterns of the first generalised spike of the SWD showed less variability (mean: 2.5 per child; SD: 2.07), mainly involved the fronto-temporal/temporal areas, and correlated poorly (<10%) with that of the pre-generalisation focal SWD. Our findings suggest that the process of generalised epileptogenesis in genetic epilepsies with electrographic "frontal absences" is far more complex than that proposed by the model for occult frontal focus with fast secondary generalisation. (Published with Supplemental data).

A hemodynamic network involving the insula, the cingulate, and the basal forebrain correlates with EEG synchronization phases of sleep instability.

The cyclic alternating pattern (CAP) encompasses the pseudoperiodic appearance of synchronized brain waves and rhythms and is considered a regulator of the nonrapid eye movement (NREM) sleep vigilance level, reflecting sleep instability. To determine the brain regions responsible for this phenomenon, we scored and analyzed sleep functional magnetic resonance imaging data acquired with simultaneous electroencephalography (EEG-fMRI). Group analysis revealed a set of brain areas showing statistically significant blood oxygen-level dependent signal correlated positively with the synchronization phase of the CAP, most prominent being the insula, the middle cingulate gyrus, and the basal forebrain. These areas may form a network acting as a synchronization pacemaker, controlling the level of NREM sleep vigilance and the sleeper's arousability.

Spatiotemporal propagation patterns of generalized ictal spikes in childhood absence epilepsy.

OBJECTIVE: This work investigates the spatial distribution in time of generalized ictal spikes in the typical absences of childhood absence epilepsy (CAE). METHODS: We studied twelve children with CAE, who had more than two typical absences during their routine video-EEG. Seizures were identified, and ictal spikes were marked over the maximum electronegative peak, clustered, waveform-averaged and spatiotemporaly analyzed in 2D electrode space. RESULTS: Consistency of spatiotemporal patterns of ictal spikes was high between the absences of the same child, but low between children. Three main discharge patterns were identified: of anterio-posterior propagation, of posterio-anterior propagation and confined to the frontal/prefrontal regions. In 4 patients, the propagation patterns transformed during the seizure into either a lateralized diminished or a non-lateralized reverse direction form. Most spikes originated fronto-temporaly, all maximized over the frontal/prefrontal electrodes and mostly decayed prefrontaly. In 4 patients, lateralized propagation patterns were identified. CONCLUSIONS: Ictal spike propagation patterns suggest that epileptogenic CAE networks are personalized, interconnect distal areas in the brain - not the entire cortex - with a tendency to generate bilateral symmetrical discharges, sometimes unsuccessfully. The transformation of propagation patterns during the seizure indicates the existence of dynamic interplay within epileptogenic networks. SIGNIFICANCE: Our results support the revised concept of ictogenesis of ILAE definition in genetic (also known as idiopathic) generalized epilepsies. Understanding the focal features in CAE avoids misdiagnosis as focal epilepsy and inappropriate treatment.

Connectivity Measures in EEG Microstructural Sleep Elements.

During Non-Rapid Eye Movement sleep (NREM) the brain is relatively disconnected from the environment, while connectedness between brain areas is also decreased. Evidence indicates, that these dynamic connectivity changes are delivered by microstructural elements of sleep: short periods of environmental stimuli evaluation followed by sleep promoting procedures. The connectivity patterns of the latter, among other aspects of sleep microstructure, are still to be fully elucidated. We suggest here a methodology for the assessment and investigation of the connectivity patterns of EEG microstructural elements, such as sleep spindles. The methodology combines techniques in the preprocessing, estimation, error assessing and visualization of results levels in order to allow the detailed examination of the connectivity aspects (levels and directionality of information flow) over frequency and time with notable resolution, while dealing with the volume conduction and EEG reference assessment. The high temporal and frequency resolution of the methodology will allow the association between the microelements and the dynamically forming networks that characterize them, and consequently possibly reveal aspects of the EEG microstructure. The proposed methodology is initially tested on artificially generated signals for proof of concept and subsequently applied to real EEG recordings via a custom built MATLAB-based tool developed for such studies. Preliminary results from 843 fast sleep spindles recorded in whole night sleep of 5 healthy volunteers indicate a prevailing pattern of interactions between centroparietal and frontal regions. We demonstrate hereby, an opening to our knowledge attempt to estimate the scalp EEG connectivity that characterizes fast sleep spindles via an "EEG-element connectivity" methodology we propose. The application of the latter, via a computational tool we developed suggests it is able to investigate the connectivity patterns related to the occurrence of EEG microstructural elements. Network characterization of specified physiological or pathological EEG microstructural elements can potentially be of great importance in the understanding, identification, and prediction of health and disease.

Semi-automatic sleep EEG scoring based on the hypnospectrogram.

BACKGROUND: Sleep EEG organization is revealed by sleep scoring, a time-consuming process based on strictly defined visual criteria. NEW METHOD: We explore the possibility of sleep scoring using the whole-night time-frequency analysis, termed hypnospectrogram, with a computer-assisted K-means clustering method. RESULTS: Hypnograms were derived from 10 whole-night sleep EEG recordings using either standard visual scoring under the Rechtshaffen and Kales criteria or semi-automated analysis of the hypnospectrogram derived from a single EEG electrode. We measured substantial agreement between the two approaches with Cohen's kappa considering all 7 stages at 0.61. COMPARISON WITH EXISTING METHODS: A number of existing automated procedures have reached the level of human inter-rater agreement using the standard criteria. However, our approach offers the scorer the opportunity to exploit the information-rich graphic representation of the whole night sleep upon which the automated method works. CONCLUSION: This work suggests that the hypnospectrogram can be used as an objective graphical rep-resentation of sleep architecture upon which sleep scoring can be performed with computer-assisted methods.

Spindle power is not affected after spontaneous K-complexes during human NREM sleep.

K-complexes and sleep spindles often grouped together characterize the second stage of NREM sleep and interest has been raised on a possible interaction of their underlying mechanisms. The reported inhibition of spindles power for about 15 seconds following evoked K-complexes has implications on their role in arousal. Our objective was to assess this inhibition following spontaneous K-complexes. We used time-frequency analysis of spontaneous K-complexes selected from whole-night EEG recordings of normal subjects. Our results show that spindles are most often observed at the positive phase following the peak of a spontaneous KC (70%). At latencies of 1-3 s following the peak of the K-complex, spindles almost disappear. Compared to long-term effects described for evoked KCs, sleep spindle power is not affected by spontaneous KCs for latencies of 5-15 s. Observation of the recurrence rate of sporadic spindles suggests that the reduction of power at 1-3 s most likely reflects a refractory period of spindles lasting for 1-2 s, rather than an effect of KCs. These results suggest that the mechanisms underlying spontaneous KCs do not affect spindle power as in the case of evoked KCs.

An intra-K-complex oscillation with independent and labile frequency and topography in NREM sleep.

NREM sleep is characterized by K-complexes (KCs), over the negative phase of which we identified brief activity in the theta range. We recorded high resolution EEG of whole-night sleep from seven healthy volunteers and visually identified 2nd and 3rd stage NREM spontaneous KCs. We identified three major categories: (1) KCs without intra-KC-activity (iKCa), (2) KCs with non-oscillatory iKCa, and (3) KCs with oscillatory iKCa. The latter group of KCs with intra-KC-oscillation (iKCo), was clustered according to the duration of the iKCo. iKCa was observed in most KCs (1150/1522, 75%). iKCos with 2, 3, and 4 waves were observed in 52% (786/1522) of KCs in respective rates of 49% (386/786), 44%, and 7%. Successive waves of iKCos showed on average a shift of their maximal amplitude in the anterio-posterior axis, while the average amplitude of the slow KC showed no spatial shift in time. The iKCo spatial shift was accompanied by transient increases in instantaneous frequency from the theta band toward the alpha band, followed by decreases to upper theta. The study shows that the KC is most often concurrently accompanied by an independent brief iKCo exhibiting topographical relocation of amplitude maxima with every consecutive peak and transient increases in frequency. The iKCo features are potentially reflecting arousing processes taking place during the KC.