Local Dosing in a 3-Mercaptopropionic Acid Chemically-Induced Epileptic Seizure Model with Microdialysis Sampling

The focus of this research was the development of an animal model for local administration of 3-mercaptopropionic acid (3-MPA) in a chemically-induced epileptic seizure model using microdialysis sampling with simultaneous electrocorticography recording (ECoG). Local administration of 3-MPA through the microdialysis probe was employed to elicit seizures in a localized brain region. Delivery of 3-MPA to the brain and changes in amino acid and catecholamine neurotransmitters were monitored. Simultaneous ECoG recordings were made using a microdialysis probe with an internal Ag/AgCl electrode. Local administration of a convulsant is important, as many clinical cases present with focal seizures. Neurochemical and electrical activity were monitored in three separate brain regions: the striatum, hippocampus, and locus coeruleus. 3-MPA was administered through the microdialysis probe in one region, while control samples were collected in the other two. These results demonstrated that unless two brain regions were connected via efferent or afferent pathways, administration of 3-MPA in one region had no neurochemical effect in the others. In the region where 3-MPA was administered, an increase in both glutamate, the main excitatory amino acid, and GABA, the main inhibitory amino acid, was seen. In addition, an increase in both dopamine and norepinephrine was seen. A multiple dosing regimen of 3-MPA was developed where 3-MPA was administered twice. These results showed that there was an attenuation in the increase of glutamate and GABA during the second administration of 3-MPA, indicating a neuronal protective mechanism taking place to decrease the effect of the second 3-MPA administration. Seizures were not detected using during local administration of 3-MPA using the microdialysis probes with an internal Ag/AgCl electrode. This was not due to the ineffectiveness of the electrodes, as they detected seizures during systemic dosing of 3-MPA. It is possible that the number of neurons excited from the local administration of 3-MPA were so limited that the signal was too small to be detected

Computational aspects of parvalbumin-positive interneuron function

The activity of neurons is dependent on the manner in which they process synaptic inputs from other cells. In the event of clustered synaptic input, neurons can respond in a nonlinear manner through synaptic and dendritic mechanisms. Such mechanisms are well established in principal excitatory neurons throughout the brain, where they increase neuronal computational ability and information storage capacity. In contrast for parvalbumin-positive (PV+) interneurons, the most common cortical class of in- hibitory interneuron, synaptic integration is thought to be either linear or sub-linear in nature, facilitating their role as mediators of precise and fast inhibition. This thesis addresses situations in which PV+ interneurons integrate synaptic inputs in a nonlinear manner, and explores the functions of this synaptic processing. First, I describe a form of cooperative supralinear synaptic integration by local excitatory inputs onto PV+ interneurons, and I extend these results to show how this augments the computational capability of PV+ cells within spiking neuron networks. I also explore the importance of polyamine-modulation of synaptic receptors in mediating sublinear synaptic integration, and discuss how this expands the array of mechanisms known to perform similar functions in PV+ cells. Finally, I present work manipulating PV+ cells experimentally during epilepsy. I consider these findings together with recent scientific advances and suggest how they account for a number of open questions and previously contradictory theories of PV+ interneuron function

Whole Brain Network Dynamics of Epileptic Seizures at Single Cell Resolution

Epileptic seizures are characterised by abnormal brain dynamics at multiple scales, engaging single neurons, neuronal ensembles and coarse brain regions. Key to understanding the cause of such emergent population dynamics, is capturing the collective behaviour of neuronal activity at multiple brain scales. In this thesis I make use of the larval zebrafish to capture single cell neuronal activity across the whole brain during epileptic seizures. Firstly, I make use of statistical physics methods to quantify the collective behaviour of single neuron dynamics during epileptic seizures. Here, I demonstrate a population mechanism through which single neuron dynamics organise into seizures: brain dynamics deviate from a phase transition. Secondly, I make use of single neuron network models to identify the synaptic mechanisms that actually cause this shift to occur. Here, I show that the density of neuronal connections in the network is key for driving generalised seizure dynamics. Interestingly, such changes also disrupt network response properties and flexible dynamics in brain networks, thus linking microscale neuronal changes with emergent brain dysfunction during seizures. Thirdly, I make use of non-linear causal inference methods to study the nature of the underlying neuronal interactions that enable seizures to occur. Here I show that seizures are driven by high synchrony but also by highly non-linear interactions between neurons. Interestingly, these non-linear signatures are filtered out at the macroscale, and therefore may represent a neuronal signature that could be used for microscale interventional strategies. This thesis demonstrates the utility of studying multi-scale dynamics in the larval zebrafish, to link neuronal activity at the microscale with emergent properties during seizures

Normal And Epilepsy-Associated Pathologic Function Of The Dentate Gyrus

The dentate gyrus plays critical roles both in cognitive processing and in regulating propagation of pathological, synchronous activity through the limbic system. The cellular and circuit mechanisms underlying these diverse functions overlap extensively. At the cellular level, the intrinsic properties of dentate granule cells combine to make these neurons fundamentally reluctant to activate, one of their hallmark traits. At the circuit level, the dentate gyrus is one of the more heavily inhibited regions of the brain, with powerful feedforward and feedback GABAergic inhibition dominating responses to afferent activation. In pathologic states such as epilepsy, disease-associated alterations within the dentate gyrus combine to compromise this circuit’s regulatory properties, culminating in a collapse of its normal function. Through the use of dynamic circuit imaging and electrophysiological brain slice recordings, pharmacology, immunohistochemistry, and a pilocarpine model of epilepsy, I characterize the emergence of dentate granule cell firing properties during brain development and then examine how the circuit’s normal activation properties become corrupted as epilepsy develops. I find that, in the perinatal brain, dentate granule cells activate in large numbers. As animals mature, these cells become less excitable and activate in extremely sparse populations in a precise, repeatable, frequency-dependent manner. This sparse activation is mediated by local circuit inhibition and not by alterations in afferent innervation of granule cells. Later, in a pilocarpine model of epilepsy, I demonstrate that normally sparse granule cell activation is massively enhanced during both epilepsy development and expression. This augmentation in excitability is mediated primarily by local disinhibition, and the mechanistic cause of this compromised inhibitory function varies over time following epileptogenic injury. My results implicate a reduction in chloride ion extrusion as a mechanism compromising inhibitory function and contributing to granule cell hyperactivation specifically during early epilepsy development. In contrast, we demonstrate that sparse dentate granule cell activation in chronically epileptic mice is rescued by glutamine application, implicating compromised GABA synthesis as a mechanism of disinhibition in chronic epilepsy. We conclude that compromised feedforward inhibition within the local circuit is the predominant mediator of the massive dentate gyrus circuit hyperactivation evident in animals during and following epilepsy development

Epileptic Seizures and the EEG

A study of epilepsy from an engineering perspective, this volume begins by summarizing the physiology and the fundamental ideas behind the measurement, analysis and modeling of the epileptic brain. It introduces the EEG and provides an explanation of the type of brain activity likely to register in EEG measurements, offering an overview of how these EEG records are and have been analyzed in the past. The book focuses on the problem of seizure detection and surveys the physiologically based dynamic models of brain activity. Finally, it addresses the fundamental question: can seizures be predicted? Based on the authors' extensive research, the book concludes by exploring a range of future possibilities in seizure prediction

Epileptic Seizures and the EEG

A study of epilepsy from an engineering perspective, this volume begins by summarizing the physiology and the fundamental ideas behind the measurement, analysis and modeling of the epileptic brain. It introduces the EEG and provides an explanation of the type of brain activity likely to register in EEG measurements, offering an overview of how these EEG records are and have been analyzed in the past. The book focuses on the problem of seizure detection and surveys the physiologically based dynamic models of brain activity. Finally, it addresses the fundamental question: can seizures be predicted? Based on the authors' extensive research, the book concludes by exploring a range of future possibilities in seizure prediction

Opportunities for improving animal welfare in rodent models of epilepsy and seizures

Animal models of epilepsy and seizures, mostly involving mice and rats, are used to understand the pathophysiology of the different forms of epilepsy and their comorbidities, to identify biomarkers, and to discover new antiepileptic drugs and treatments for comorbidities. Such models represent an important area for application of the 3Rs (replacement, reduction and refinement of animal use). This report provides background information and recommendations aimed at minimising pain, suffering and distress in rodent models of epilepsy and seizures in order to improve animal welfare and optimise the quality of studies in this area. The report includes practical guidance on principles of choosing a model, induction procedures, in vivo recordings, perioperative care, welfare assessment, humane endpoints, social housing, environmental enrichment, reporting of studies and data sharing. In addition, some model-specific welfare considerations are discussed, and data gaps and areas for further research are identified. The guidance is based upon a systematic review of the scientific literature, survey of the international epilepsy research community, consultation with veterinarians and animal care and welfare officers, and the expert opinion and practical experience of the members of a Working Group convened by the United Kingdom's National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs)

Effects of intracranial stimulation and the involvement of the human parahippocampal cortex in perception

How the human brain translates photons hitting the retina into conscious perception remains an open question. Throughout the medial temporal lobe (MTL), there are neurons (called concept cells) that change their firing rate when that neuron's preferred concept, e.g., a specific person or object, is seen. The firing rate of concept cells is correlated with perception. Nevertheless, it remains unclear whether or to what extent concept cells are involved in perceptogenesis, i.e., the creation of conscious percepts. Inferring from studies in monkeys, concept-specific neurons involved in perceptogenesis would be expected along the ventral and dorsal stream of visual processing (also called the what and where pathway, respectively). Various regions that are part of the dorsal stream are connected to the parahippocampal cortex (PHC), a region within the MTL. Compared to other MTL regions, lower selectivity, the absence of multimodal responses, and especially the shorter response latencies do not exclude an involvement of the PHC in perceptogenesis. In fact, damage to the parahippocampal place area (PPA, a part of the PHC) results in topographical disorientation. The goal of this thesis is to test the involvement of the PHC in perception by using electrical stimulation during a forced-choice categorization task involving landscapes versus animals. First, we determined effective parameters for intracranial stimulation of brain tissue in epilepsy patients implanted with depth-electrodes for seizure monitoring. We investigated the effects of amplitude, phase width, frequency, and pulse-train duration on neuronal firing, the local field potential (LFP), and behavioral responses to evoked percepts. Frequency and charge per phase were the most influential parameters on all three signals. Both parameters showed a positive effect on event-related potentials (ERPs) in the LFP. Higher frequencies (especially around 200 Hz) lead to a short-term inhibition of neuronal firing, while higher charge per phase can have an inhibitory or excitatory effect on neuronal firing. All parameters had a positive effect on the reports of evoked percepts; on reports of phosphenes in response to stimulating close to the optic radiation as well as on reports of auditory verbal hallucinations in response to stimulating Heschl's gyrus. Using functional magnetic resonance imaging (fMRI), we found that the PPA, i.e., the part of the PHC that is most selective towards images of landscapes, is rather small (up to 1‰ of total brain volume per hemisphere) with varying degrees of hemispheric laterality. Stimulating the PHC outside of the PPA - using a 100 ms high-frequency pulse train delivered at the natural response latency of the PHC - had no effect on categorizing landscapes. However, stimulating inside the PPA, close to the peak activation of the fMRI cluster, resulted in a 7% to 10% increase in landscape responses to ambiguous stimuli. Furthermore, stimulating the PPA also led to an increase in behavioral response time, especially to images with a predominant landscape component. None of our patients reported visual hallucinations of places or scenes in response to our stimulation protocols. Our data suggests that the PPA is involved in the perceptogenesis of landscapes at a stage that does not reach awareness, while the rest of the PHC is unlikely to be involved in perceptogenesis, at least not as it pertains to the perception of landscapes or animals. We also developed an online spike sorting algorithm and an adaptive screening procedure for concept cells to pave the way for new paradigms involving informed feedback

Changes in neuronal firing and synchrony precede recruitment of mesial temporal networks into generalizing seizures

Despite extensive study, the mechanisms underlying seizure generation and propagation are poorly understood. One approach is to study changes in the neuronal activity (of inhibitory and excitatory subpopulations) that occur during the recruitment of networks into a propagating seizure, to gain insight into mechanisms by which seizures spread across the brain. Recent work, comparing intra- and extracelluar recordings in ex-vivo preparations of human neocortex has implicated a failure in feed-forward inhibition underlying the spread of seizure. However, direct in-vivo study of inhibitory and excitatory population dynamics in the neocortex is difficult, due to an inability to separate single neuron activity into excitatory and inhibitory subpopulations. In the mesial temporal lobe (MTL) it is considerably easier to isolate these subpopulations, and several studies in the rodent MTL have, indeed, demonstrated an intricate spatiotemporal interplay between inhibitory and excitatory neuron firing and their corresponding synchrony to local field potentials during the transition to seizure. While this work suggests potential mechanisms for network recruitment into seizure, no direct observations have been made in the MTL of epileptic patients. This report provides methods to prolong the longevity of single neuron recordings in the human MTL. Using these recordings, evidence is presented that supports the hypothesis that recruitment of MTL networks into seizures of neocortical origin is preceded by specific spatiotemporal increases in synchrony. In detail, within the MTL there is a decrease in inhibitory interneuron firing that coincides with the inhibitory population becoming more coherent to their local field potentials. This increased synchrony between neurons and the local field occurs at frequencies similar to those of regional synchrony between MTL networks and the seizure focus. These results suggest a mechanism by which downstream networks are prepared for recruitment into generalizing seizures. Interestingly, these spatiotemporal changes occur prior to the first electrographic manifestation of seizure in the brain, implying that in addition to their role in seizure propagation, changes in interneuron firing and interneuron-field synchrony in the MTL may be reflective of early seizure activity in other brain structures as well, and may thus be a useful tool in developing improved early detection algorithms.Ph.D., Biomedical Engineering -- Drexel University, 201