Paolo Belardinelli

TMS-EEG Signatures of GABAergic Neurotransmission in the Human Cortex

Combining transcranial magnetic stimulation (TMS) and electroencephalography (EEG) constitutes a powerful tool to directly assess human cortical excitability and connectivity. TMS of the primary motor cortex elicits a sequence of TMS-evoked EEG potentials (TEPs). It is thought that inhibitory neurotransmission through GABA-A receptors (GABAAR) modulates early TEPs (<50 ms after TMS), whereas GABA-B receptors (GABABR) play a role for later TEPs (at ∼100 ms after TMS). However, the physiological underpinnings of TEPs have not been clearly elucidated yet. Here, we studied the role of GABAA/B-ergic neurotransmission for TEPs in healthy subjects using a pharmaco-TMS-EEG approach. In Experiment 1, we tested the effects of a single oral dose of alprazolam (a classical benzodiazepine acting as allosteric-positive modulator at α1, α2, α3, and α5 subunit-containing GABAARs) and zolpidem (a positive modulator mainly at the α1 GABAAR) in a double-blind, placebo-controlled, crossover study. In Experiment 2, we tested the influence of baclofen (a GABABR agonist) and diazepam (a classical benzodiazepine) versus placebo on TEPs. Alprazolam and diazepam increased the amplitude of the negative potential at 45 ms after stimulation (N45) and decreased the negative component at 100 ms (N100), whereas zolpidem increased the N45 only. In contrast, baclofen specifically increased the N100 amplitude. These results provide strong evidence that the N45 represents activity of α1-subunit-containing GABAARs, whereas the N100 represents activity of GABABRs. Findings open a novel window of opportunity to study alteration of GABAA-/GABAB-related inhibition in disorders, such as epilepsy or schizophrenia.

Characterization of GABAB-receptor mediated neurotransmission in the human cortex by paired-pulse TMS-EEG

GABAB-receptor (GABABR) mediated inhibition is important in regulating neuronal excitability. The paired-pulse transcranial magnetic stimulation (TMS) protocol of long-interval intracortical inhibition (LICI) likely reflects this GABABergic inhibition. However, this view is based on indirect evidence from electromyographic (EMG) studies. Here we combined paired-pulse TMS with simultaneous electroencephalography (paired-pulse TMS-EEG) and pharmacology to directly investigate mechanisms of LICI at the cortical level. We tested the effects of a conditioning stimulus (CS100) applied 100ms prior to a test stimulus (TS) over primary motor cortex on TS-evoked EEG-potentials (TEPs). Healthy subjects were given a single oral dose of baclofen, a GABABR agonist, or diazepam, a positive modulator at GABAARs, in a placebo-controlled, pseudo-randomized double-blinded crossover study. LICI was quantified as the difference between paired-pulse TEPs (corrected for long-lasting EEG responses by the conditioning pulse) minus single-pulse TEPs. LICI at baseline (i.e. pre-drug intake) was characterized by decreased P25, N45, N100 and P180 and increased P70 TEP components. Baclofen resulted in a trend towards the enhancement of LICI of the N45 and N100, and significantly enhanced LICI of the P180. In contrast, diazepam consistently suppressed LICI of late potentials (i.e. N100, P180), without having an effect on LICI of earlier (i.e. P25, N45 and P70) potentials. These findings demonstrate for the first time directly at the system level of the human cortex that GABABR-mediated cortical inhibition contributes to LICI, while GABAAR-mediated inhibition occludes LICI. Paired-pulse TMS-EEG allows investigating cortical GABABR-mediated inhibition more directly and specifically than hitherto possible, and may thus inform on network abnormalities caused by disordered inhibition, e.g. in patients with schizophrenia or epilepsy.

Source reconstruction accuracy of MEG and EEG Bayesian inversion approaches.

Electro- and magnetoencephalography allow for non-invasive investigation of human brain activation and corresponding networks with high temporal resolution. Still, no correct network detection is possible without reliable source localization. In this paper, we examine four different source localization schemes under a common Variational Bayesian framework. A Bayesian approach to the Minimum Norm Model (MNM), an Empirical Bayesian Beamformer (EBB) and two iterative Bayesian schemes (Automatic Relevance Determination (ARD) and Greedy Search (GS)) are quantitatively compared. While EBB and MNM each use a single empirical prior, ARD and GS employ a library of anatomical priors that define possible source configurations. The localization performance was investigated as a function of (i) the number of sources (one vs. two vs. three), (ii) the signal to noise ratio (SNR; 5 levels) and (iii) the temporal correlation of source time courses (for the cases of two or three sources). We also tested whether the use of additional bilateral priors specifying source covariance for ARD and GS algorithms improved performance. Our results show that MNM proves effective only with single source configurations. EBB shows a spatial accuracy of few millimeters with high SNRs and low correlation between sources. In contrast, ARD and GS are more robust to noise and less affected by temporal correlations between sources. However, the spatial accuracy of ARD and GS is generally limited to the order of one centimeter. We found that the use of correlated covariance priors made no difference to ARD/GS performance.

Closed-Loop Neuroscience and Non-Invasive Brain Stimulation: A Tale of Two Loops

Closed-loop neuroscience is receiving increasing attention with recent technological advances that enable complex feedback loops to be implemented with millisecond resolution on commodity hardware. We summarize emerging conceptual and methodological frameworks that are available to experimenters investigating a “brain in the loop” using non-invasive brain stimulation and briefly review the experimental and therapeutic implications. We take the view that closed-loop neuroscience in fact deals with two conceptually quite different loops: a “brain-state dynamics” loop, used to couple with and modulate the trajectory of neuronal activity patterns, and a “task dynamics” loop, that is the bidirectional motor-sensory interaction between brain and (simulated) environment, and which enables goal-directed behavioral tasks to be incorporated. Both loops need to be considered and combined to realize the full experimental and therapeutic potential of closed-loop neuroscience.

Steady-state responses in MEG demonstrate information integration within but not across the auditory and visual senses

To form a unified percept of our environment, the human brain integrates information within and across the senses. This MEG study investigated interactions within and between sensory modalities using a frequency analysis of steady-state responses that are elicited time-locked to periodically modulated stimuli. Critically, in the frequency domain, interactions between sensory signals are indexed by crossmodulation terms (i.e. the sums and differences of the fundamental frequencies). The 3 × 2 factorial design, manipulated (1) modality: auditory, visual or audiovisual (2) steady-state modulation: the auditory and visual signals were modulated only in one sensory feature (e.g. visual gratings modulated in luminance at 6 Hz) or in two features (e.g. tones modulated in frequency at 40 Hz & amplitude at 0.2 Hz). This design enabled us to investigate crossmodulation frequencies that are elicited when two stimulus features are modulated concurrently (i) in one sensory modality or (ii) in auditory and visual modalities. In support of within-modality integration, we reliably identified crossmodulation frequencies when two stimulus features in one sensory modality were modulated at different frequencies. In contrast, no crossmodulation frequencies were identified when information needed to be combined from auditory and visual modalities. The absence of audiovisual crossmodulation frequencies suggests that the previously reported audiovisual interactions in primary sensory areas may mediate low level spatiotemporal coincidence detection that is prominent for stimulus transients but less relevant for sustained SSR responses. In conclusion, our results indicate that information in SSRs is integrated over multiple time scales within but not across sensory modalities at the primary cortical level.

Corticomuscular Coherence Is Tuned to the Spontaneous Rhythmicity of Speech at 2–3 Hz

Human speech features rhythmicity that frames distinctive, fine-grained speech patterns. Speech can thus be counted among rhythmic motor behaviors that generally manifest characteristic spontaneous rates. However, the critical neural evidence for tuning of articulatory control to a spontaneous rate of speech has not been uncovered. The present study examined the spontaneous rhythmicity in speech production and its relationship to cortex–muscle neurocommunication, which is essential for speech control. Our MEG results show that, during articulation, coherent oscillatory coupling between the mouth sensorimotor cortex and the mouth muscles is strongest at the frequency of spontaneous rhythmicity of speech at 2–3 Hz, which is also the typical rate of word production. Corticomuscular coherence, a measure of efficient cortex–muscle neurocommunication, thus reveals behaviorally relevant oscillatory tuning for spoken language.

Effects of the Selective α5-GABAAR Antagonist S44819 on Excitability in the Human Brain: A TMS-EMG and TMS-EEG Phase I Study.

Alpha-5 gamma-aminobutyric acid type A receptors (α5-GABAARs) are located extrasynaptically, regulate neuronal excitability through tonic inhibition, and are fundamentally important for processes such as plasticity and learning. For example, pharmacological blockade of α5-GABAAR in mice with ischemic stroke improved recovery of function by normalizing exaggerated perilesional α5-GABAAR-dependent tonic inhibition. S44819 is a novel competitive selective antagonist of the α5-GABAAR at the GABA-binding site. Pharmacological modulation of α5-GABAAR-mediated tonic inhibition has never been investigated in the human brain. Here, we used transcranial magnetic stimulation (TMS) to test the effects of a single oral dose of 50 and 100 mg of S44819 on electromyographic (EMG) and electroencephalographic (EEG) measures of cortical excitability in 18 healthy young adults in a randomized, double-blinded, placebo-controlled, crossover phase I study. A dose of 100 mg, but not 50 mg, of S44819 decreased active motor threshold, the intensity needed to produce a motor evoked potential of 0.5 mV, and the amplitude of the N45, a GABAAergic component of the TMS-evoked EEG response. The peak serum concentration of 100 mg S44819 correlated directly with the decrease in N45 amplitude. Short-interval intracortical inhibition, a TMS–EMG measure of synaptic GABAAergic inhibition, and other components of the TMS-evoked EEG response remained unaffected. These findings provide first time evidence that the specific α5-GABAAR antagonist S44819 reached human cortex to impose an increase in cortical excitability. These data warrant further development of S44819 in a human clinical trial to test its efficacy in enhancing recovery of function after ischemic stroke. SIGNIFICANCE STATEMENT The extrasynaptic α-5 gamma-aminobutyric acid type A receptor (α5-GABAAR) regulates neuronal excitability through tonic inhibition in the mammalian brain. Tonic inhibition is important for many fundamental processes such as plasticity and learning. Pharmacological modulation of α5-GABAAR-mediated tonic inhibition has never been investigated in the human brain. This study demonstrates that S44819, a selective α5-GABAAR antagonist, increases cortical excitability in healthy human subjects, as indicated by specific markers of transcranial magnetic stimulation-induced muscle and brain responses measured by electromyography and electroencephalography. Our findings imply that tonic inhibition in human cortex can be modified effectively and that this modification can be quantified with noninvasive brain stimulation methods. The actions of S44819 may be suitable to improve plasticity and learning.

Modulation of alpha oscillations in insular cortex reflects the threat of painful stimuli

Pain is a sensory and emotional experience that involves numerous brain areas. Among these areas the insular cortex has been shown to be involved in the expectation and processing of pain. Alpha power modulation has been associated with the experience of pain. The aim of this study was to test the hypothesis that the threat of a painful stimulus affects alpha rhythm oscillation in the insular cortex and to find the time intervals during which the insular cortex is most active. We used a beamforming method in the frequency domain to estimate alpha power associated with source activity during psychologically different conditions, namely a sequence of nonpainful somatosensory stimuli (non-threatening condition) and a sequence of nonpainful stimuli randomly intermixed with painful stimuli (threatening condition). The results revealed that the anterior insula alone was involved during the threat of painful stimuli. Conversely, the posterior insula - as well as other brain areas such as SII - was involved in the processing of somatosensory stimuli regardless their painfulness. Additionally, the involvement of the anterior insula should not be accounted for by fear, arousal, habituation effect or by the occurrence of randomly interleaved different stimuli, but it is likely to be related mainly to expectancy mechanisms enhancing activity of specific neuronal populations.

Weighted Phase Lag Index and Graph Analysis: Preliminary Investigation of Functional Connectivity during Resting State in Children

Resting state functional connectivity of MEG data was studied in 29 children (9-10 years old). The weighted phase lag index (WPLI) was employed for estimating connectivity and compared to coherence. To further evaluate the network structure, a graph analysis based on WPLI was used to determine clustering coefficient (C) and betweenness centrality (BC) as local coefficients as well as the characteristic path length (L) as a parameter for global interconnectedness. The networks modular structure was also calculated to estimate functional segregation. A seed region was identified in the central occipital area based on the power distribution at the sensor level in the alpha band. WPLI reveals a specific connectivity map different from power and coherence. BC and modularity show a strong level of connectedness in the occipital area between lateral and central sensors. C shows different isolated areas of occipital sensors. Globally, a network with the shortest L is detected in the alpha band, consistently with the local results. Our results are in agreement with findings in adults, indicating a similar functional network in children at this age in the alpha band. The integrated use of WPLI and graph analysis can help to gain a better description of resting state networks.

Plasticity of premotor cortico-muscular coherence in severely impaired stroke patients with hand paralysis

Motor recovery in severely impaired stroke patients is often very limited. To refine therapeutic interventions for regaining motor control in this patient group, the functionally relevant mechanisms of neuronal plasticity need to be detected. Cortico-muscular coherence (CMC) may provide physiological and topographic insights to achieve this goal. Synchronizing limb movements to motor-related brain activation is hypothesized to reestablish cortico-motor control indexed by CMC. In the present study, right-handed, chronic stroke patients with right-hemispheric lesions and left hand paralysis participated in a four-week training for their left upper extremity. A brain-robot interface turned event-related beta-band desynchronization of the lesioned sensorimotor cortex during kinesthetic motor-imagery into the opening of the paralyzed hand by a robotic orthosis. Simultaneous MEG/EMG recordings and individual models from MRIs were used for CMC detection and source reconstruction of cortico-muscular connectivity to the affected finger extensors before and after the training program. The upper extremity-FMA of the patients improved significantly from 16.23 ± 6.79 to 19.52 ± 7.91 (p = 0.0015). All patients showed significantly increased CMC in the beta frequency-band, with a distributed, bi-hemispheric pattern and considerable inter-individual variability. The location of CMC changes was not correlated to the severity of the motor impairment, the motor improvement or the lesion volume. Group analysis of the cortical overlap revealed a common feature in all patients following the intervention: a significantly increased level of ipsilesional premotor CMC that extended from the superior to the middle and inferior frontal gyrus, along with a confined area of increased CMC in the contralesional premotor cortex. In conclusion, functionally relevant modulations of CMC can be detected in patients with long-term, severe motor deficits after a brain-robot assisted rehabilitation training. Premotor beta-band CMC may serve as a biomarker and therapeutic target for novel treatment approaches in this patient group.