Burst Discharges Research Articles

Enhanced burst discharges in the CA1 area of the immature versus adult hippocampus: patterns and cellular mechanisms.

Burst discharges in the immature brain may contribute to its enhanced seizure susceptibility. The cellular mechanisms underlying burst discharges in the CA1 area of the immature versus adult hippocampus were investigated with simultaneous whole-cell and field-potential recordings. When GABAA receptors were blocked pharmacologically, bursts in CA1 were either graded or all-or-none (or mixed) as a function of electrical stimulation intensity. Most CA1 minislices from immature rats displayed all-or-none or mixed bursts, whereas the slices from adult rats predominantly elicited graded bursts. The frequency and amplitude of spontaneous excitatory postsynaptic currents (sEPSCs) were greater in CA1 pyramidal cells from the immature than the adult slices. The developmental differences in CA1 bursting were also detected in slices adjusted for maturational changes in brain volume (i.e., 350 µm thick for immature vs. 450 µm thick for adult rats). Neither N-methyl-d-aspartate (NMDA) nor group I metabotropic glutamate (mGlu1) receptor antagonists blocked the network-driven bursts in immature CA1, but an α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor blocker abolished them. Robust excitatory postsynaptic potentials (EPSPs) occurred after bursts in some immature CA1 slices (23%) but never in slices from the adult. The input-output (amount of current injected vs. number of action potentials generated) relationship was markedly greater in CA1 pyramidal cells in the immature compared with the adult hippocampus. These data suggest that the CA1 area of the immature brain is capable of generating network-driven bursts, which declines in adult rats. The increased propensity of burst generation in immature CA1 appears to involve a greater AMPA receptor-mediated synaptic network and an increased intrinsic spike-generating ability.NEW & NOTEWORTHY Burst discharges in the developing brain can provide valuable insights into epileptogenesis. We show that the immature hippocampal CA1 area is capable of generating all-or-none (i.e., network) bursts, which transitions to graded (i.e., nonnetwork) bursts in the mature brain via both synaptic and intrinsic mechanisms. Our results provide new clues to help understand possible mechanisms that may be shared in the immature and epileptic brain and how the normal brain becomes seizure prone (i.e., epileptogenesis).

Formation and consumption of HO2 radicals in ns pulse O2–He plasmas over a liquid water surface

Hydroperoxyl (HO2) radicals are an important precursor in the formation of hydrogen peroxide (H2O2), a key species in plasma-liquid interactions, such that their formation and consumption pathways need to be understood. In this work, the generation and decay of HO2 have been studied in a controlled environment, in ns pulse discharge O2–He plasmas in contact with a liquid water surface. For this, time-resolved, absolute number densities of HO2 in O2–He mixtures excited by a repetitive ns pulse discharge are measured in situ by cavity ring down spectroscopy (CRDS). The discharge cell with external electrodes to generate the plasma and a water reservoir are integrated into the CRDS cavity. The high-reflectivity cavity mirrors are purged with helium to protect them from water vapor condensation. The experimental results are obtained at near room temperature, both during the discharge pulse burst and in the afterglow. The HO2 number density is inferred from the CRDS data using a spectral model exhibiting good agreement with previous measurements of absolute HO2 absorption cross sections. HO2 is generated during the discharge burst and decays in the afterglow between the bursts. The HO2 number density is also measured vs. the O2 fraction in the mixture. Comparison with the kinetic modeling predictions demonstrates good agreement with the data and identifies the dominant HO2 generation and decay processes. HO2 in the plasma is formed predominantly by the recombination of H atoms, generated by the electron impact dissociation of water vapor, with O2 molecules. Reactions with O atoms and hydroxyl (OH) radicals are among the main HO2 decay processes in the afterglow. HO2 is also detected when O2 is not present in the mixture. In this case, it is generated primarily by the recombination of OH radicals, via the formation of H2O2. The results demonstrate that CRDS can also be used for HO2 and other plasma chemical reaction product measurements in atmospheric pressure plasma jets impinging on a liquid water surface in ambient air.

Preictal phase detection on EEG signals using hybridized machine learning classifiers with a novel feature selection procedure based GAs and ICOMP

Epilepsy is the fourth most common neurological disorder, which affects the brain and brings out frequent seizures. They are bursts of electrical discharge that can cause a wide range of symptoms such as distraction or involuntary spasms involving the whole body. Preictal phase carries some important features related to seizures which can be found before seizure onset. Hence, this study introduces an efficient hybrid training procedure for machine learning (ML) classifiers that are able to classify Electroencephalogram (EEG) signals for the accurate detection of preictal phase. Essentially, the proposed approach consists of two stages: feature extraction and model estimation with feature selection. In this approach, while the feature extraction is executed by using wavelet transform, the model estimation is performed by hybrid ML classifiers. Essentially, this approach integrates the training mechanism with a novel feature subset and model selection procedure based on the Information Complexity Criteria (ICOMP) and Genetic Algorithms. For preictal phase detection application, the CHB-MIT Scalp EEG dataset was analyzed by both the proposed and traditional approaches. From the analysis results, it can be concluded that the hybrid ML classifiers not only produce robust models in the context of model information complexity, but also provide superior performance outputs than the classical approaches with respect to validity and reliability, over test datasets.

Time-resolved CO2, CO, and N2 vibrational population measurements in Ns pulse discharge plasmas

Time-resolved CO2 and N2 vibrational populations and translational-rotational temperature are measured in a CO2–N2 plasma sustained by a ns pulse discharge burst in plane-to-plane geometry. Time-resolved, absolute number density of CO generated in the plasma is also inferred from the experimental data. CO2 and CO vibrational populations are measured by mid-IR, tunable quantum cascade laser absorption spectroscopy, and N2 vibrational populations are measured by the ns broadband vibrational CARS. Transient excitation of N2 and CO2 asymmetric stretch vibrational energy modes is detected during the discharge burst. The time-resolved rate of CO generation does not correlate with N2 or CO2(ν 3) vibrational temperatures, indicating that CO2 dissociation via the vibrational excitation is insignificant at the present conditions. The rate of CO generation decreases gradually during the discharge burst. The estimated specific energy cost of the CO product is close to that of N atoms in pure nitrogen, measured previously at similar operating conditions. Comparison of the experimental data with the kinetic modeling analysis indicates that CO2 dissociation in collisions with electronically excited N2 molecules is the dominant channel of CO generation at the present conditions, although the inferred CO yield in these processes is significantly lower than 1. The effect of vibrational energy transfer between N2 and CO2 on the plasma chemical processes is insignificant. The kinetic model underpredicts a rapid reduction of the N2 and CO2(ν 3) vibrational temperatures during the later half of the discharge burst and in the afterglow. V–T relaxation of N2 by N and O atoms generated in the ns pulse discharge plasma does not affect the vibrational relaxation rate in a significant way. However, rapid V–T relaxation of CO2 by O atoms has a significant effect on the relaxation rate. The difference between the experimental data and the modeling predictions may be due to the unknown scaling of the CO2–O V–T rates with the vibrational quantum number.

Focal seizures are organized by feedback between neural activity and ion concentration changes.

Human and animal EEG data demonstrate that focal seizures start with low-voltage fast activity, evolve into rhythmic burst discharges and are followed by a period of suppressed background activity. This suggests that processes with dynamics in the range of tens of seconds govern focal seizure evolution. We investigate the processes associated with seizure dynamics by complementing the Hodgkin-Huxley mathematical model with the physical laws that dictate ion movement and maintain ionic gradients. Our biophysically realistic computational model closely replicates the electrographic pattern of a typical human focal seizure characterized by low voltage fast activity onset, tonic phase, clonic phase and postictal suppression. Our study demonstrates, for the first time in silico, the potential mechanism of seizure initiation by inhibitory interneurons via the initial build-up of extracellular K+ due to intense interneuronal spiking. The model also identifies ionic mechanisms that may underlie a key feature in seizure dynamics, that is, progressive slowing down of ictal discharges towards the end of seizure. Our model prediction of specific scaling of inter-burst intervals is confirmed by seizure data recorded in the whole guinea pig brain in vitro and in humans, suggesting that the observed termination pattern may hold across different species. Our results emphasize ionic dynamics as elementary processes behind seizure generation and indicate targets for new therapeutic strategies.

Optimal selection of multipolar electrode configurations for nerve burst detection.

Nerve cuff electrodes are commonly used for neural stimulation and recording applications. Usually, these electrodes are composed of a limited set of metal rings, disposed around the nerve. Although widely used, this technology may be insufficient to record and stimulate in a more selective manner. Higher resolution electrodes, usually composed of a matrix of independent contact points, have been proposed in this sense. These electrodes allow for the exploration of a wide variety of bipolar or multipolar setups, for selective recording and stimulation. In this study, we propose a method to optimally select such multipolar setups and to quantitatively evaluate the performance of a multi-contact neural organic electrode (OE) in recording burst discharges from the rat's phrenic nerve. A 16-channel OE was wrapped around the phrenic nerve (studied electrode) and a suction electrode was applied to the cut-end of the same nerve (gold standard electrode). Analysis of all possible combinations of bipoles and tripoles from the OE were carried out to assess the improvement in the recording performance, measured as the signal-to-noise ratio, compared to the gold standard. The results showed that the bipolar and tripolar configuration significantly increased the overall recording performance. Such configurations are therefore essential to improve nerve burst detection.

The genesis and functional consequences of cortico-subthalamic beta augmentation and excessive subthalamic burst discharges after dopaminergic deprivation

The cardinal electrophysiological signs in Parkinson's disease (PD) include augmented beta oscillations in the motor cortex-subthalamic nucleus (MC-STN) axis and excessive burst discharges in STN. We have shown that excessive STN burst discharges have a direct causal relation with the locomotor deficits in PD. To investigate the correlation between the two cardinal signs, we characterized the courses of development of the electrophysiological abnormalities in the hemiparkinsonian rat model. The loss of dopaminergic neurons develops fast, and is histologically completed within 4–7 days of the lesion. The increase in STN burst discharges is limited to the lesioned side, and follows a very similar course. In contrast, beta augmentation has a bilateral presentation, and requires 14–21 days for full development. Behaviorally, the gross locomotor deficits in open field test and limb akinesia in stepping test match the foregoing fast and slow time courses, respectively. A further look into the spike entrainment shows that the oscillations in local field potential (LFP) of the MC effectively entrain the multi-unit (MU) spikes of MC, STN and entopeduncular nucleus (EPN), a rat homolog of human globus pallidus interna (GPi), whereas the LFP of STN or EPN (GPi) cannot entrain the spikes in MC. We conclude that excessive STN burst discharges are a direct consequence, whereas beta augmentation is probably a secondary or adaptive changes in the cortico-subcortical re-entrant loops, to dopaminergic deprivation. Beta augmentation is therefore not so consistently present as excessive STN burst discharges, but could signal more delicate derangements at the level of cortical programming in PD.

Deep brain stimulation rectifies the noisy cortex and irresponsive subthalamus to improve parkinsonian locomotor activities

The success of deep brain stimulation (DBS) therapy indicates that Parkinson’s disease is a brain rhythm disorder. However, the manifestations of the erroneous rhythms corrected by DBS remain to be established. We found that augmentation of α rhythms and α coherence between the motor cortex (MC) and the subthalamic nucleus (STN) is characteristically prokinetic and is decreased in parkinsonian rats. In multi-unit recordings, movement is normally associated with increased changes in spatiotemporal activities rather than overall spike rates in MC. In parkinsonian rats, MC shows higher spike rates at rest but less spatiotemporal activity changes upon movement, and STN burst discharges are more prevalent, longer lasting, and less responsive to MC inputs. DBS at STN rectifies the foregoing pathological MC-STN oscillations and consequently locomotor deficits, yet overstimulation may cause behavioral restlessness. These results indicate that delicate electrophysiological considerations at both cortical and subcortical levels should be exercised for optimal DBS therapy.

RNA Interference Via shRNA Decreases the Number of Transcripts of Scn8a in Glutamatergic Interneurons in the preBötC

Rhythmic neural activity that drives the inspiratory phase of the breathing cycle in mammals emanates from glutamatergic interneurons of the pre‐Bötzinger Complex (preBötC). A longstanding question in neuroscience is: what are the underlying mechanisms for inspiratory rhythmogenesis? One proposed mechanism involves the persistent sodium current (INaP), which is the ionic mechanism that underlies the voltage‐dependent bursting‐pacemaker activity of neurons whose rhythmic burst discharges are putatively obligatory for inspiratory rhythm generation. Specifically in brain tissues, the Scn8a gene codes for Nav1.6 ion channels, and those ion channels give rise to INaP.Different genetic tools can help us to understand the role of specific molecules in the generation of motor behaviors, such as breathing. The short hairpin RNA (shRNA) is an artificial RNA molecule, delivered through adeno‐associated virus (AAV), that can inhibit gene expression and promote long‐term knock‐down of the targeted gene.Here we administered Cre‐dependent shRNA to knock‐down Scn8a (thus Nav1.6) in Cre‐expressing glutamatergic neurons in preBötC and quantified mRNA per glutamatergic neuron. We used adult Vglut2Cre mice (10‐14 weeks old) and injected 50 nL of AAVs carrying the shRNA. We used in situ hybridization (RNAscope® Fluorescent Assay) to label mRNA of Scn8a and to identify transduced glutamatergic neurons in the preBötC for RNA quantification at 2 and 6 weeks after injection. Samples were visualized under the Four‐Channel, Upright Confocal Microscope Body Based on Cerna® System (ThorLabs), and images were acquired using the ThorImage®LS Data Acquisition Software, at 40x magnification. The amount of RNA per cell was quantified using the open‐source software Quantitative Pathology(QuPath). We obtained the mean of the number of transcripts per cell ± the standard deviation (SD), and unpaired t‐test was used to evaluate the statistical difference between the shRNA injected mice and their control group (mice whose viral vector contained a non‐targeting shRNA sequence).Our results show that at 2 weeks, the control group had 27.5±9.5 RNA/neuron, whereas the shRNA‐injected had 12.8±3.5, which represents a 53% of decrease in the number of transcripts (P<0.0001). A similar pattern was observed after 6 weeks of the injection in a different cohort, where control animals showed 35.4±6.1 RNA/cells, and the shRNA group only had 24.1±5.2, representing 32% drop in the transcripts for that group (P<0.0001). This initial protocol allowed us to verify that the gene expression has been knocked down at critical time points, encouraging further studies using shRNA to evaluate whether attenuation of Nav1.6 expression in rhythmogenic preBötC neurons impairs breathing, which could resolve the longstanding question regarding the putatively rhythmogenic role of INaP.

Generation, breakdown and responses of ATP in the carotid body

Purinergic signaling involving adenosine triphosphate (ATP) acting on P2X2/3 receptors modulates physiological carotid body (CB) afferent discharge and chemoreflex activation however, the upregulation of P2X3 receptors on petrosal sensory neurons is partly responsible for aberrant CB tonicity and hyperreflexia, and the increase in blood pressure in the Spontaneously Hypertensive rat (SHR). The enhanced chemoreflex occurs prior to the onset of hypertension and elevated CB activity is thought to contribute to the underlying pathophysiology of the disease but the exact mechanisms remain unclear. We hypothesize that there is greater release of ATP and/or decreased ATP breakdown in the pre‐hypertensive SHR CB compared with that in the Wistar rat, and this evokes an increase in the chemoreflex response and subsequent activation of the sympathetic nervous system. We performed three studies. (i) Using digital droplet PCR (ddPCR), we measured mRNA expression levels of enzymes involved in the breakdown of ATP in CBs extracted from male 4‐week‐old prehypertensive SHR and Wistar rats; these included: ectonucleotide pyrophosphatase/phosphodiesterase 1‐3 (Enpp1‐3), ectonucleoside triphosphate diphosphohydrolase 2‐3 (Entpd2‐3), and ecto‐5’‐nucleotidase (Nt5e). (ii) We quantified ATP release from CBs in vitro during baseline conditions and during cyanide‐evoked ATP release using a colorimetric assay (213‐579‐1 ‐ Millipore), comparing between male 4‐week‐old pre‐hypertensive SHR and Wistar rats. (iii) In an in situ working heart‐brainstem preparation of 4‐week‐old Wistar rats, simultaneous recordings were made from phrenic (PN), hypoglossal (HN), recurrent laryngeal (RLN), abdominal (ABN), and thoracic sympathetic nerves (tSNA). The peripheral chemoreceptors were stimulated with either potassium cyanide (KCN 20‐100 μL, 0.04%) or ATP at varying concentrations (10‐100 µL, 50 µM ‐ 2 mM) delivered via the internal carotid artery. In whole CB extracts, ddPCR revealed increased Enpp2‐3 and Nt5e expression (p<0.05) in pre‐hypertensive SHRs relative to Wistar rats suggesting greater capacity to break down ATP in the pre‐hypertensive SHR CB. There were no differences in the expressions of Enpp1 or Entpd2‐3 between rat strains. However, the amount of ATP released from the CB in both baseline and during cyanide stimulation was two‐fold greater in the pre‐hypertensive SHR (P<0.05). Stimulation of the CB with KCN evoked hyperpnoea, bradycardia and sympathoexcitation at all doses. In contrast, CB injection of low dose ATP increased PN rate with little effect on tSNA, whereas higher doses induced apnea accompanied by repetitive burst discharge of post‐inspiratory activity in the RLN with tonic elevations in the discharge of both ABN and tSNA. In sum, despite the upregulation of several ATP degrading enzymes in the CBs of the SHR, CB release of ATP is higher in this species. We hypothesize that the increase in ATP release in the SHR CB drives the CB hyperexcitability and that the shift towards an upregulation of ATP degrading enzymes even prior to the development of hypertension is compensatory to oppose the increase in ATP signaling. The dose‐dependent effect of exogenous ATP at supraphysiological levels evoking greater CB‐mediated expiratory and sympathetic outflow in the Wistar rat mimics aspects of pathophysiological CB chemoreflex signaling in the SHR.

Glioma facilitates the epileptic and tumor-suppressive gene expressions in the surrounding region

Patients with glioma often demonstrate epilepsy. We previously found burst discharges in the peritumoral area in patients with malignant brain tumors during biopsy. Therefore, we hypothesized that the peritumoral area may possess an epileptic focus and that biological alterations in the peritumoral area may cause epileptic symptoms in patients with glioma. To test our hypothesis, we developed a rat model of glioma and characterized it at the cellular and molecular levels. We first labeled rat C6 glioma cells with tdTomato, a red fluorescent protein (C6-tdTomato), and implanted them into the somatosensory cortex of VGAT-Venus rats, which specifically expressed Venus, a yellow fluorescent protein in GABAergic neurons. We observed that the density of GABAergic neurons was significantly decreased in the peritumoral area of rats with glioma compared with the contralateral healthy side. By using a combination technique of laser capture microdissection and RNA sequencing (LCM-seq) of paraformaldehyde-fixed brain sections, we demonstrated that 19 genes were differentially expressed in the peritumoral area and that five of them were associated with epilepsy and neurodevelopmental disorders. In addition, the canonical pathways actively altered in the peritumoral area were predicted to cause a reduction in GABAergic neurons. These results suggest that biological alterations in the peritumoral area may be a cause of glioma-related epilepsy.

Ignition dynamics of radio frequency discharge in atmospheric pressure cascade glow discharge

A cascade glow discharge in atmospheric helium was excited by a microsecond voltage pulse and a pulse-modulated radio frequency (RF) voltage, in which the discharge ignition dynamics of the RF discharge burst was investigated experimentally. The spatio-temporal evolution of the discharge, the ignition time and optical emission intensities of plasma species of the RF discharge burst were investigated under different time intervals between the pulsed voltage and RF voltage in the experiment. The results show that by increasing the time interval between the pulsed discharge and RF discharge burst from 5 μs to 20 μs, the ignition time of the RF discharge burst is increased from 1.6 μs to 2.0 μs, and the discharge spatial profile of RF discharge in the ignition phase changes from a double-hump shape to a bell-shape. The light emission intensity at 706 nm and 777 nm at different time intervals indicates that the RF discharge burst ignition of the depends on the number of residual plasma species generated in the pulsed discharges.

Rhythmic firing of neurons in the medulla of conscious freely behaving rats: rhythmic coupling with baroreceptor input.

Recent investigations emphasized the importance of neural control of cardiovascular adjustments in complex behaviors, including stress, exercise, arousal, sleep-wake states, and different tasks. Baroreceptor feedback is an essential component of this system acting on different time scales from maintaining stable levels of cardiovascular parameters on the long-term to rapid alterations according to behavior. The baroreceptor input is essentially rhythmic, reflecting periodic fluctuations in arterial blood pressure. Cardiac rhythm is a prominent feature of the autonomic control system, present on different levels, including neuron activity in central circuits. The mechanism of rhythmic entrainment of neuron firing by the baroreceptor input was studied in great detail under anesthesia, but recordings of sympathetic-related neuron firing in freely moving animals remain extremely scarce. In this study, we recorded multiple single neuron activity in the reticular formation of the medulla in freely moving rats during natural behavior. Neurons firing in synchrony with the cardiac rhythm were detected in each experiment (n = 4). In agreement with prior observations in anesthetized cats, we found that neurons in this area exhibited high neuron-to-neuron variability and temporal flexibility in their coupling to cardiac rhythm in freely moving rats, as well. This included firing in bursts at multiples of cardiac cycles, but not directly coupled to the heartbeat, supporting the concept of baroreceptor input entraining intrinsic neural oscillations rather than imposing a rhythm of solely external origin on these networks. It may also point to a mechanism of maintaining the basic characteristics of sympathetic neuron activity, i.e., burst discharge and cardiac-related rhythmicity, on the background of behavior-related adjustments in their firing rate.

Time-resolved measurements of HO2 radical in a heated plasma flow reactor

Time-resolved, absolute HO2 number density in diluted H2O2-Ar, CH4O2-Ar, and C2H4O2-Ar mixtures excited by a repetitive ns pulse discharge in a heated plasma flow reactor is measured by Cavity Ringdown Spectroscopy (CRDS). The experimental results are obtained at T = 300–600 K and P = 130 Torr, both during the discharge pulse burst and in the afterglow. The HO2 number density is inferred from the CRDS data using a spectral model exhibiting good agreement with previous measurements of absolute HO2 absorption cross sections. In the room-temperature H2O2 mixture, as well as in CH4O2 and C2H4O2 mixtures over the entire temperature range studied, HO2 is generated only during the discharge burst and decays in the afterglow. However, in the H2O2 mixture at elevated temperatures, T = 400–600 K, HO2 persists in the afterglow up to 10 ms after the discharge burst, comparable with the flow residence time in the reactor. Comparison with kinetic modeling shows that the sustained reactivity after the source of radicals is turned off is due to a chain propagation/ hydrogen oxidation process, which dominates the radical recombination reactions. The kinetic modeling predictions are in good agreement with the relative HO2 number density measured in all three mixtures, although the model underpredicts the absolute number densities in H2O2 at T = 400–600 K by up to a factor of two. Detection of the sustained low-temperature reactivity in H2O2, initiated by the radical generation in the plasma, suggests that the plasma excitation may also affect kinetics of oxidation and reforming of fuels exhibiting low-temperature chemistry below hot ignition point.

Selective stabilization of the intermediate inactivated Na+ channel by the new-generation anticonvulsant rufinamide

Na+ channels undergo multiple inactivated states with different kinetics, which set the refractory period of neuronal discharges, but isolating the intermediate inactivated state has been challenging. Most classical Na+channel-inhibiting anticonvulsants bind to the fast inactivated state to reduce Na+currents and cellular excitability. These anticonvulsants have the slow binding kinetics and thus necessitate long depolarization for drug action, a “use-dependent” effect sparing most normal activities. Rufinamide is a new anticonvulsant targeting Na+channels, and has a therapeutic effect on Lennox-Gastaut syndrome (LGS) which is refractory to classicalNa+channel inhibitors. The efficacy on LGS, whose epileptiform discharges largely involve short depolarization or bursts, is primarily due to the very fast binding kinetics of rufinamide. Could the very fast kinetics of rufinamide lead to indiscriminate inhibition of neuronal activities ? Onhippocampal neurons from male and female mice, wefound that rufinamide most effectively shifts the Na+channel inactivation curve if the inactivating pulse is 1 s, rather than 0.1 or 18 s, in duration. Rufinamide also shows a maximal slowing effect on the recovery kinetics from the inactivation driven by modest depolarization (e.g. −60 mV) of intermediate length (e.g. 50–300 ms). Consistently, rufinamide selectively inhibits the burst discharges at 50–300 ms on a plateau of ∼−60 mV. This is mechanistically ascribable to selective binding of rufinamide to an intermediate inactivated state withan apparent dissociation constantof ∼40 μM. Being the first molecule embodying the evasive transitional gating state, rufinamide could have a unique anti-seizure profile with a novel form of use-dependent action.

Brivaracetam prevents the development of epileptiform activity when administered early after cortical neurotrauma in rats.

There is no effective therapy to prevent the development of posttraumatic epilepsy (PTE). Recently, we reported that administration of the antiseizure medication (ASM) levetiracetam (LEV) shortly after trauma prevented the development of epileptiform activity in two experimental models of neurotrauma. However, the time window for effective intervention with LEV may be too narrow for most clinical settings. Using the controlled cortical impact (CCI) injury model, the current study tested whether early administration of brivaracetam (BRV), an ASM with 20 times the affinity of LEV for the SV2A synaptic vesicle protein, could improve upon the antiepileptogenic action observed with LEV. Rats (postnatal day [P] 24-32) subjected to CCI injury were given a single dose of BRV (21 or 100mg/kg, i.p.) at one of three post-injury time points: immediately (0-2minutes), 30 minutes, or 60minutes. Control animals received only vehicle (0.9% saline). Posttraumatic electrographic epileptiform activity was assayed ex vivo from coronal neocortical slices collected proximal to the injury (four per rat) 3-4weeks after injury. In this model, ictal-like burst discharges occur spontaneously or can be evoked in an "all or none" manner with applied electrical stimulation within the first 2weeks after injury. A single dose of BRV administered to rats up to 60minutes after traumatic brain injury (TBI) significantly reduced the development of posttraumatic epileptiform activity by (1) inhibiting the development of both evoked and spontaneous epileptiform activity, (2) raising the threshold for stimulus-evoked epileptiform discharges, and (3) reducing the intensity of epileptiform bursts that arise after cortical neurotrauma. Clinically there has been little success preventing the development of posttraumatic epilepsy. The results of this study support the hypothesis that early intervention with BRV has the potential to prevent or reduce posttraumatic epileptogenesis, and that there may be a limited time window for successful prophylactic intervention.

Measurements of atoms and metastable species in N2 and H2–N2 nanosecond pulse plasmas

Time-resolved, absolute number densities of metastable N2(A3Σ u +, v = 0, 1) molecules, ground state N2 and H atoms, and rotational–translational temperature have been measured by tunable diode laser absorption spectroscopy and two-photon absorption laser-induced fluorescence in diffuse N2 and N2–H2 plasmas during and after a nanosecond pulse discharge burst. Comparison of the measurement results with the kinetic modeling predictions, specifically the significant reduction of the N2(A3Σ u +) populations and the rate of N atom generation during the burst, suggests that these two trends are related. The slow N atom decay in the afterglow, on a time scale longer than the discharge burst, demonstrates that the latter trend is not affected by N atom recombination, diffusion to the walls, or convection with the flow. This leads to the conclusion that the energy pooling in collisions of N2(A3Σ u +) molecules is a major channel of N2 dissociation in electric discharges where a significant fraction of the input energy goes to electronic excitation of N2. Additional measurements in a 1% H2–N2 mixture demonstrate a further significant reduction of N2(A3Σ u +, v = 0, 1) populations, due to the rapid quenching by H atoms accumulating in the plasma. Comparison with the modeling predictions suggests that the N2(A3Σ u +) molecules may be initially formed in the highly vibrationally excited states. The reduction of the N2(A3Σ u +) number density also diminishes the contribution of the energy pooling process into N2 dissociation, thus reducing the N atom number density. The rate of N atom generation during the burst also decreases, due to its strong coupling to N2(A3Σ u +, v) populations. On the other hand, the rate of H atom generation, produced predominantly by the dissociative quenching of the excited electronic states of N2 by H2, remains about the same during the burst, resulting in a nearly linear rise in the H atom number density. Comparison of the kinetic model predictions with the experimental results suggests that the yield of H atoms during the quenching of the excited electronic state of N2 by molecular H2 is significantly less than 100%. The present results quantify the yield of N and H atoms in high-pressure H2–N2 plasmas, which have significant potential for ammonia generation using plasma-assisted catalysis.

Magnetically responsive hydrophobic pockets for on–off drug release

The vast majority of drugs available on the market are hydrophobic compounds. As a result, their poor water solubility can critically compromise the overall absorption of these drugs by patients. Although numerous different strategies have been developed to improve their bioavailability, the controlled delivery of these drugs is still a challenge. In this sense, stimuli-responsive hydrogels could be a solution to improve administration and stable release. However, the strategies required to render hydrogels hydrophobic mostly rely on weak hydrophobic interactions, which can lead to disassembly of the system and undesired burst discharge. Accordingly, the on-demand release of poorly water-soluble drugs is still a major milestone in this field. To circumvent these setbacks, we present for the first time a hydrophobic, magnetically responsive hydrogel based on the diaminotriazine (DAT) skeleton. The versatility of DATs in terms of H-bond formation and metal complexation simultaneously endows the hydrogel with hydrophobicity and magnetic responsiveness, thus allowing both the efficient loading and on–off release of a model hydrophobic drug as well as of a hydrophobic bioactive substance. Theoretical calculations further suggested stable formation of DAT aggregates that operate as efficient hydrophobic cavities or ‘pockets’ for these compounds. Moreover, these materials were found to be suitable for enhancing the viability and growth of cell lines often used as in vitro models of neuronal function and differentiation. The present magnetic hydrogels hold interesting potential for tissue engineering as biocompatible scaffolds for the on-demand release of hydrophobic compounds, which remains the focus of current research efforts.

Theta-Burst Stimulation of Primary Afferents Drives Long-Term Potentiation in the Spinal Cord and Persistent Pain via α2δ-1-Bound NMDA Receptors.

Long-term potentiation (LTP) and long-term depression (LTD) in the spinal dorsal horn reflect activity-dependent synaptic plasticity and central sensitization in chronic pain. Tetanic high-frequency stimulation is commonly used to induce LTP in the spinal cord. However, primary afferent nerves often display low-frequency, rhythmic bursting discharges in painful conditions. Here, we determined how theta-burst stimulation (TBS) of primary afferents impacts spinal cord synaptic plasticity and nociception in male and female mice. We found that TBS induced more LTP, whereas tetanic stimulation induced more LTD, in mouse spinal lamina II neurons. TBS triggered LTP, but not LTD, in 50% of excitatory neurons expressing vesicular glutamate transporter-2 (VGluT2). By contrast, TBS induced LTD and LTP in 12-16% of vesicular GABA transporter (VGAT)-expressing inhibitory neurons. Nerve injury significantly increased the prevalence of TBS-induced LTP in VGluT2-expressing, but not VGAT-expressing, lamina II neurons. Blocking NMDARs, inhibiting α2δ-1 with gabapentin, or α2δ-1 knockout abolished TBS-induced LTP in lamina II neurons. Also, disrupting the α2δ-1-NMDAR interaction with α2δ-1Tat peptide prevented TBS-induced LTP in VGluT2-expressing neurons. Furthermore, TBS of the sciatic nerve induced long-lasting allodynia and hyperalgesia in wild-type, but not α2δ-1 knockout, mice. TBS significantly increased the α2δ-1-NMDAR interaction and synaptic trafficking in the spinal cord. In addition, treatment with NMDAR antagonists, gabapentin, or α2δ-1Tat peptide reversed TBS-induced pain hypersensitivity. Therefore, TBS-induced primary afferent input causes a neuropathic pain-like phenotype and LTP predominantly in excitatory dorsal horn neurons via α2δ-1-dependent NMDAR activation. α2δ-1-bound NMDARs may be targeted for reducing chronic pain development at the onset of tissue/nerve injury.SIGNIFICANCE STATEMENT Spinal dorsal horn synaptic plasticity is a hallmark of chronic pain. Although sensory nerves display rhythmic bursting discharges at theta frequencies during painful conditions, the significance of this naturally occurring firing activity in the induction of spinal synaptic plasticity is largely unknown. In this study, we found that theta-burst stimulation (TBS) of sensory nerves induced LTP mainly in excitatory dorsal horn neurons and that the prevalence of TBS-induced LTP was potentiated by nerve injury. This TBS-driven synaptic plasticity required α2δ-1 and its interaction with NMDARs. Furthermore, TBS of sensory nerves induced persistent pain, which was maintained by α2δ-1-bound NMDARs. Thus, TBS-induced LTP at primary afferent-dorsal horn neuron synapses is an appropriate cellular model for studying mechanisms of chronic pain.

Intrinsic and synaptic mechanisms controlling the expiratory activity of excitatory lateral parafacial neurones of rats.

Active expiration is essential for increasing pulmonary ventilation during high chemical drive (hypercapnia). The lateral parafacial (pFL ) region, which contains expiratory neurones, drives abdominal muscles during active expiration in response to hypercapnia. However, the electrophysiological properties and synaptic mechanisms determining the activity of pFL expiratory neurones, as well as the specific conditions for their emergence, are not fully understood. Using whole cell electrophysiology and single cell quantitative RT-PCR techniques, we describe the intrinsic electrophysiological properties, the phenotype and the respiratory-related synaptic inputs to the pFL expiratory neurones, as well as the mechanisms for the expression of their expiratory activity under conditions of hypercapnia-induced active expiration, using in situ preparations of juvenile rats. We also evaluated whether these neurones possess intrinsic CO2 /[H+ ] sensitivity and burst generating properties. GABAergic and glycinergic inhibition during inspiration and expiration suppressed the activity of glutamatergic pFL expiratory neurones in normocapnia. In hypercapnia, these neurones escape glycinergic inhibition and generate burst discharges at the end of expiration. Evidence for the contribution of post-inhibitory rebound, CaV 3.2 isoform of T-type Ca2+ channels and intracellular [Ca2+ ] is presented. Neither intrinsic bursting properties, mediated by persistent Na+ current, nor CO2 /[H+ ] sensitivity or expression of CO2 /[H+ ] sensitive ion channels/receptors (TASK or GPR4) were observed. On the other hand, hyperpolarisation-activated cyclic nucleotide-gated and twik-related K+ leak channels were recorded. Post-synaptic disinhibition and the intrinsic electrophysiological properties of glutamatergic neurones play important roles in the generation of the expiratory oscillations in the pFL region during hypercapnia in rats. KEY POINTS: Hypercapnia induces active expiration in rats and the recruitment of a specific population of expiratory neurones in the lateral parafacial (pFL ) region. Post-synaptic GABAergic and glycinergic inhibition both suppress the activity of glutamatergic pFL neurones during inspiratory and expiratory phases in normocapnia. Hypercapnia reduces glycinergic inhibition during expiration leading to burst generation by pFL neurones; evidence for a contribution of post-inhibitory rebound, voltage-gated Ca2+ channels and intracellular [Ca2+ ] is presented. pFL glutamatergic expiratory neurones are neither intrinsic burster neurones, nor CO2 /[H+ ] sensors, and do not express CO2 /[H+ ] sensitive ion channels or receptors. Post-synaptic disinhibition and the intrinsic electrophysiological properties of glutamatergic neurones both play important roles in the generation of the expiratory oscillations in the pFL region during hypercapnia in rats.