Charles Quairiaux

A mouse model for studying large-scale neuronal networks using EEG mapping techniques

Human functional imaging studies are increasingly focusing on the identification of large-scale neuronal networks, their temporal properties, their development, and their plasticity and recovery after brain lesions. A method targeting large-scale networks in rodents would open the possibility to investigate their neuronal and molecular basis in detail. We here present a method to study such networks in mice with minimal invasiveness, based on the simultaneous recording of epicranial EEG from 32 electrodes regularly distributed over the head surface. Spatiotemporal analysis of the electrical potential maps similar to human EEG imaging studies allows quantifying the dynamics of the global neuronal activation with sub-millisecond resolution. We tested the feasibility, stability and reproducibility of the method by recording the electrical activity evoked by mechanical stimulation of the mystacial vibrissae. We found a series of potential maps with different spatial configurations that suggested the activation of a large-scale network with generators in several somatosensory and motor areas of both hemispheres. The spatiotemporal activation pattern was stable both across mice and in the same mouse across time. We also performed 16-channel intracortical recordings of the local field potential across cortical layers in different brain areas and found tight spatiotemporal concordance with the generators estimated from the epicranial maps. Epicranial EEG mapping thus allows assessing sensory processing by large-scale neuronal networks in living mice with minimal invasiveness, complementing existing approaches to study the neurophysiological mechanisms of interaction within the network in detail and to characterize their developmental, experience-dependent and lesion-induced plasticity in normal and transgenic animals.

PV plasticity sustained through D1/5 dopamine signaling required for long-term memory consolidation.

Long-term consolidation of memories depends on processes occurring many hours after acquisition. Whether this involves plasticity that is specifically required for long-term consolidation remains unclear. We found that learning-induced plasticity of local parvalbumin (PV) basket cells was specifically required for long-term, but not short/intermediate-term, memory consolidation in mice. PV plasticity, which involves changes in PV and GAD67 expression and connectivity onto PV neurons, was regulated by cAMP signaling in PV neurons. Following induction, PV plasticity depended on local D1/5 dopamine receptor signaling at 0–5 h to regulate its magnitude, and at 12–14 h for its continuance, ensuring memory consolidation. D1/5 dopamine receptor activation selectively induced DARPP-32 and ERK phosphorylation in PV neurons. At 12–14 h, PV plasticity was required for enhanced sharp-wave ripple densities and c-Fos expression in pyramidal neurons. Our results reveal general network mechanisms of long-term memory consolidation that requires plasticity of PV basket cells induced after acquisition and sustained subsequently through D1/5 receptor signaling.

The physiological plausibility of time-varying Granger-causal modeling: normalization and weighting by spectral power.

Time-varying connectivity methods are increasingly used to study directed interactions between brain regions from electrophysiological signals. These methods often show good results in simulated data but it is unclear to what extent connectivity results obtained from real data are physiologically plausible. Here we introduce a benchmark approach using multichannel somatosensory evoked potentials (SEPs) measured across rat cortex, where the structural and functional connectivity is relatively simple and well-understood. Rat SEPs to whisker stimulation are exclusively initiated by contralateral primary sensory cortex (S1), at known latencies, and with activity spread from S1 to specific cortical regions. This allows for a comparison of time-varying connectivity measures according to fixed criteria. We thus evaluated the performance of time-varying Partial Directed Coherence (PDC) and the Directed Transfer Function (DTF), comparing row- and column-wise normalization and the effect of weighting by the power spectral density (PSD). The benchmark approach revealed clear differences between methods in terms of physiological plausibility, effect size and temporal resolution. The results provide a validation of time-varying directed connectivity methods in an animal model and suggest a driving role for ipsilateral S1 in the later part of the SEP. The benchmark SEP dataset is made freely available.

Long-Term Plasticity in Mouse Sensorimotor Circuits after Rhythmic Whisker Stimulation

Mice actively explore their environment by rhythmically sweeping their whiskers. As a consequence, neuronal activity in somatosensory pathways is modulated by the frequency of whisker movement. The potential role of rhythmic neuronal activity for the integration and consolidation of sensory signals, however, remains unexplored. Here, we show that a brief period of rhythmic whisker stimulation in anesthetized mice resulted in a frequency-specific long-lasting increase in the amplitude of somatosensory-evoked potentials in the contralateral primary somatosensory (barrel) cortex. Mapping of evoked potentials and intracortical recordings revealed that, in addition to potentiation in layers IV and II/III of the barrel cortex, rhythmic whisker stimulation induced a decrease of somatosensory-evoked responses in the supragranular layers of the motor cortex. To assess whether rhythmic sensory input-based plasticity might arise in natural settings, we exposed mice to environmental enrichment. We found that it resulted in somatosensory-evoked responses of increased amplitude, highlighting the influence of previous sensory experience in shaping sensory responses. Importantly, environmental enrichment-induced plasticity occluded further potentiation by rhythmic stimulation, indicating that both phenomena share common mechanisms. Overall, our results suggest that natural, rhythmic patterns of whisker activity can modify the cerebral processing of sensory information, providing a possible mechanism for learning during sensory perception.

Functional development of large-scale sensorimotor cortical networks in the brain.

Large-scale neuronal networks integrating several cortical areas mediate the complex functions of the brain such as sensorimotor integration. Little is known about the functional development of these networks and the maturational processes by which distant networks become functionally connected. We addressed this question in the postnatal rat sensorimotor system. Using epicranial multielectrode grids that span most of the cortical surface and intracortical electrodes, we show that sensory evoked cortical responses continuously maturate throughout the first 3 weeks with the strongest developmental changes occurring in a very short time around postnatal day 13 (P13). Before P13, whisker stimulation evokes slow, initially surface-negative activity restricted mostly to the lateral parietal area of the contralateral hemisphere. In a narrow time window of ∼48 h around P13, a new early, sharp surface-positive component emerges that coincides with subsequent propagation of activity to sensory and motor areas of both hemispheres. Our data show that this new component developing at the end of the second week corresponds principally to functional maturation of the supragranular cortical layers and appears to be crucial for the functional associations in the large-scale sensorimotor cortical network. It goes along with the onset of whisking behavior, as well as major synaptic and functional changes within the S1 cortex that are known to develop during this period.

Multi-modal assessment of long-term erythropoietin treatment after neonatal hypoxic-ischemic injury in rat brain.

Erythropoietin (EPO) has been recognized as a neuroprotective agent. In animal models of neonatal brain injury, exogenous EPO has been shown to reduce lesion size, improve structure and function. Experimental studies have focused on short course treatment after injury. Timing, dose and length of treatment in preterm brain damage remain to be defined. We have evaluated the effects of high dose and long-term EPO treatment in hypoxic-ischemic (HI) injury in 3 days old (P3) rat pups using histopathology, magnetic resonance imaging (MRI) and spectroscopy (MRS) as well as functional assessment with somatosensory-evoked potentials (SEP). After HI, rat pups were assessed by MRI for initial damage and were randomized to receive EPO or vehicle. At the end of treatment period (P25) the size of resulting cortical damage and white matter (WM) microstructure integrity were assessed by MRI and cortical metabolism by MRS. Whisker elicited SEP were recorded to evaluate somatosensory function. Brains were collected for neuropathological assessment. The EPO treated animals did not show significant decrease of the HI induced cortical loss at P25. WM microstructure measured by diffusion tensor imaging was improved and SEP response in the injured cortex was recovered in the EPO treated animals compared to vehicle treated animals. In addition, the metabolic profile was less altered in the EPO group. Long-term treatment with high dose EPO after HI injury in the very immature rat brain induced recovery of WM microstructure and connectivity as well as somatosensory cortical function despite no effects on volume of cortical damage. This indicates that long-term high-dose EPO induces recovery of structural and functional connectivity despite persisting gross anatomical cortical alteration resulting from HI.

Functional Deficit and Recovery of Developing Sensorimotor Networks following Neonatal Hypoxic–Ischemic Injury in the Rat

Neonatal hypoxia-ischemia (HI) is the most important cause of brain injury in the newborn. Here we studied structural alterations and functional perturbations of developing large-scale sensorimotor cortical networks in a rat model of moderate HI at postnatal day 3 (P3). At the morphological level, HI led to a disorganized barrel pattern in the somatosensory cortex without detectable histological changes in the motor cortex. Functional effects were addressed by means of epicranial mapping of somatosensory-evoked potentials (SEPs) during the postischemic recovery period. At P10, SEPs were immature and evoked activity was almost restricted to the somatosensory and motor cortices of the contralateral hemisphere. Peak and topographic analyses of epicranial potentials revealed that responses were profoundly depressed in both sensory and motor areas of HI-lesioned animals. At the end of the postnatal period at P21, responses involved networks in both hemispheres. SEP amplitude was still depressed in the injured sensory region, but it completely recovered in the motor area. These results suggest a process of large-scale network plasticity in sensorimotor circuits after perinatal ischemic injury. The model provides new perspectives for investigating the temporal and spatial characteristics of the recovery process following HI and eventually developing therapeutic interventions.

Dynamic connectivity among cortical layers in local and large-scale sensory processing

Cortical processing of sensory stimuli typically recruits multiple areas, but how each area dynamically incorporates activity from other areas is not well understood. We investigated interactions between cortical columns of bilateral primary sensory regions (S1s) in rats by recording local field potentials and multi‐unit activity simultaneously in both S1s with electrodes positioned at each cortical layer. Using dynamic connectivity analysis based on Granger‐causal modeling, we found that, shortly after whisker stimulation (< 10 ms), contralateral S1 (cS1) already relays activity to granular and infragranular layers of S1 in the other hemisphere, after which cS1 shows a pattern of within‐column interactions that directs activity upwards toward superficial layers. This pattern of predominant upward driving was also observed in S1 ipsilateral to stimulation, but at longer latencies. In addition, we found that interactions between the two S1s most strongly target granular and infragranular layers. Taken together, the results suggest a possible mechanism for how cortical columns integrate local and large‐scale neocortical computation by relaying information from deeper layers to local processing in superficial layers.

Whole-scalp EEG mapping of somatosensory evoked potentials in macaque monkeys

High-density scalp EEG recordings are widely used to study whole-brain neuronal networks in humans non-invasively. Here, we validate EEG mapping of somatosensory evoked potentials (SSEPs) in macaque monkeys (Macaca fascicularis) for the long-term investigation of large-scale neuronal networks and their reorganisation after lesions requiring a craniotomy. SSEPs were acquired from 33 scalp electrodes in five adult anaesthetized animals after electrical median or tibial nerve stimulation. SSEP scalp potential maps were identified by cluster analysis and identified in individual recordings. A distributed, linear inverse solution was used to estimate the intracortical sources of the scalp potentials. SSEPs were characterised by a sequence of components with unique scalp topographies. Source analysis confirmed that median nerve SSEP component maps were in accordance with the somatotopic organisation of the sensorimotor cortex. Most importantly, SSEP recordings were stable both intra- and interindividually. We aim to apply this method to the study of recovery and reorganisation of large-scale neuronal networks following a focal cortical lesion requiring a craniotomy. As a prerequisite, the present study demonstrated that a 300-mm2 unilateral craniotomy over the sensorimotor cortex necessary to induce a cortical lesion, followed by bone flap repositioning, suture and gap plugging with calcium phosphate cement, did not induce major distortions of the SSEPs. In conclusion, SSEPs can be successfully and reproducibly recorded from high-density EEG caps in macaque monkeys before and after a craniotomy, opening new possibilities for the long-term follow-up of the cortical reorganisation of large-scale networks in macaque monkeys after a cortical lesion.

Electrophysiological evidence for the development of a self-sustained large-scale epileptic network in the kainate mouse-model of temporal lobe epilepsy

Most research on focal epilepsy focuses on mechanisms of seizure generation in the primary epileptic focus (EF). However, neurological deficits that are not directly linked to seizure activity and that may persist after focus removal are frequent. The recruitment of remote brain regions of an epileptic network (EN) is recognized as a possible cause, but a profound lack of experimental evidence exists concerning their recruitment and the type of pathological activities they exhibit. We studied the development of epileptic activities at the large-scale in male mice of the kainate model of unilateral temporal lobe epilepsy using high-density surface EEG and multiple-site intracortical recordings. We show that, along with focal spikes and fast ripples that remain localized to the injected hippocampus (i.e., the EF), a subpopulation of spikes that propagate across the brain progressively emerges even before the expression of seizures. The spatiotemporal propagation of these generalized spikes (GSs) is highly stable within and across animals, defining a large-scale EN comprising both hippocampal regions and frontal cortices. Interestingly, GSs are often concomitant with muscular twitches. In addition, while fast ripples are, as expected, highly frequent in the EF, they also emerge in remote cortical regions and in particular in frontal regions where GSs propagate. Finally, we demonstrate that these remote interictal activities are dependent on the focus in the early phase of the disease but continue to be expressed after focus silencing at later stages. Our results provide evidence that neuronal networks outside the initial focus are progressively altered during epileptogenesis. SIGNIFICANCE STATEMENT It has long been held that the epileptic focus is responsible for triggering seizures and driving interictal activities. However, focal epilepsies are associated with heterogeneous symptoms, calling into question the concept of a strictly focal disease. Using the mouse model of hippocampal sclerosis, this work demonstrates that focal epilepsy leads to the development of pathological activities specific to the epileptic condition, notably fast ripples, that appear outside of the primary epileptic focus. Whereas these activities are dependent on the focus early in the disease, focus silencing fails to control them in the chronic stage. Thus, dynamical changes specific to the epileptic condition are built up outside of the epileptic focus along with disease progression, which provides supporting evidence for network alterations in focal epilepsy.