Neurocysticercosis is a disease in which larvae of the tapeworm, Taenia solium, infect the central nervous system of humans. Seizures are the most common symptom of NCC, occurring in between 70 % and 90 % of all symptomatic NCC cases. Neurocysticercosis impacts heavily on the quality of life of patients, and further presents a significant drain on the economic resources of endemic countries. Despite its considerable global impact, the molecular and cellular mechanisms underlying seizures in neurocysticercosis remain largely unknown. In this thesis I have explored novel models for neurocysticercosis by combining mouse hippocampal organotypic brain slice cultures with various preparations of a model parasite, Taenia crassiceps. Utilising these models, I first explored, using patch clamp and local field potential electrophysiology, how Taenia larval extracts directly affect neuronal excitability. I report that extracts of Taenia crassiceps resulted in a significant acute excitation of neurons and triggered seizure-like events in brain slices. Further investigation revealed that this excitation was mediated by the activation of glutamate receptors and that, indeed, the larvae of both Taenia crassiceps and Taenia solium contain and produce levels of glutamate sufficient to explain this effect. Chronic exposure of brain slices to intact, living, larvae did not, however, result in any changes in network excitability. Next, I investigate whether Taenia larvae produce acetylcholinesterases, as these enzymes have the potential to affect neuronal signaling by digesting the neurotransmitter acetylcholine. Ellman's assays, in situ acetylcholinesterase activity assays, and patch clamp electrophysiology reveal that both Taenia crassiceps and Taenia solium larvae produce acetylcholinesterases and that the activity of Taenia acetylcholinesterases is sufficient to digest acetylcholine at a concentration that alters neuronal signaling. Finally, I explore the effect that Taenia larval extracts have on the innate immune cells of the brain, as the responses of these cells can also alter neuronal excitability. Through the measurement of brain slice cytokine release using enzyme-linked immunosorbent assays, I discover that Taenia crassiceps extracts have robust antiinflammatory effects, which involve lipid, protein, and glycan elements. This thesis presents novel findings that reveal ways in which Taenia larvae interact with both neuronal and nonneuronal resident brain cells. It further delves into how these interactions could contribute to seizure generation in neurocysticercosis and proposes some potential new therapeutic approaches to treat seizures in neurocysticercosis.

We investigated whether chronic administration of guanidinoethane sulfonate, an inhibitor of taurine uptake, could modify the antiepileptic actions of phenobarbital and phenytoin on maximal electroshock seizures in mice. Treatment with 1% guanidinoethane sulfonate decreased the taurine concentration in the brain to 76% of the control value. Under these conditions, neither the severity of tonic convulsions of maximal electroshock seizures nor the threshold for tonic extension caused by electroshock was altered. However, treatment with guanidinoethane sulfonate lessened the antiepileptic actions of phenobarbital and phenytoin on electroshock seizures. The brain concentrations of phenobarbital and phenytoin were unaltered by administration of guanidinoethane sulfonate. The brain concentrations of guanidinoethane sulfonate and total guanidino compounds were unchanged by the injection of either phenobarbital or phenytoin. It is suggested that the observed loss of anticonvulsive potency of phenobarbital and phenytoin may have been related to the decrease in taurine concentration produced by guanidinoethane sulfonate.

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Thesis (LL. M.)--University of Toronto, 2005. / Includes bibliographical references (leaves 88-95).

Made available in DSpace on 2015-07-22T20:50:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-04-29 / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Trinomys yonenagae (TY) é um roedor fossorial das dunas da Caatinga, pertencente à família Echimydae e conhecido localmente como rabo de facho. A observação de que alguns indivíduos dessa espécie animal apresentam crises convulsivas espontâneas estimulou-nos ao estudo de algumas características de seu sistema nervoso que pudessem estar subjacentes a este quadro. Através de técnicas de eletrofisiologia in vitro, com a utilização de protocolos de indução de hiperexcitabilidade e de estudo da potenciação a longo prazo, procuramos identificar o padrão eletrofisiológico da circuitaria hipocampal do TY e compará-lo ao de ratos Wistar. Adicionalmente, procuramos também verificar como animais da espécie TY se comportam em relação a dois modelos clássicos de indução de epilepsia: método do abrasamento amigdaliano e aplicação sistêmica de pilocarpina. No protocolo de potássio alto, observamos em TY uma maior sensibilidade ao aumento gradual de potássio no líquido cefalorraquiano artificial (LCRa). No protocolo de adição de antagonista do receptor GABAA (bicuculina) no LCRa, não observamos quaisquer diferenças significativas nos registros extracelulares entre TY e Wistar. No protocolo da ausência de magnésio no LCRa, ambas as espécies apresentaram atividade epileptiforme espontânea, e quando submetidas a estimulação elétrica, as respostas em ambas as espécies não diferiram estatisticamente. No estudo da potenciação a longo prazo, observamos que, embora as médias dos declives normalizados em TY terem se apresentado sempre inferiores às de Wistar após o estímulo de alta frequência, estes valores entre as duas espécies não diferiram estatisticamente. Os dados eletrencefalográficos e comportamentais foram similares entre TY e Wistar nos dois modelos de indução de epilepsia, com exceção da dose de pilocarpina utilizada para elicitar o status epilepticus em TY, que foi menor. A caracterização eletrofisiológica e os resultados obtidos mediante os modelos de epilepsia são contribuições interessantes para o conhecimento do sistema nervoso do TY e revelam a importância para futuros trabalhos nesta espécie para aquisição de novos conhecimentos que podem estar envolvidos na gênese das crises convulsivas. / Trinomys yonenagae (TY) is a fossorial rodent dweller of sand dunes of the Caatinga, pertaining to Echimydae family and known locally as rabo de facho. The observation of some animals of this species show spontaneous seizures stimulated us the study of some characteristics of its nervous system that could to be subjacents to this condition. Through techniques of eletrophysiology in vitro, with the utilization of protocols of induction of hiperexcitability and study of the long-term potentiation (LTP), we aimed at to identifying the eletrophysiologic pattern of hippocampal circuitry of the TY and comparing it to Wistar rats. Additionaly, we also aimed at verifying how animals of the TY species behave regarding two classic models of epilepsy induction: amygdala kindling and systemic aplication of pilocarpine. In the high potassium protocol, we observed in TY a higher sensibility to gradual increase of potassium in the artificial cerebrospinal fluid (ACSF). In the protocol of addition of antagonist of GABAA receptor (bicuculine) in the ACSF, we did not observe any significative differences in the extracellular records between TY and Wistar. In the absence magnesium protocol in the ACSF, both species showed spontaneous epileptiform activity, and when both species were submited to electric stimulation, their responses did not differ statisticaly. In the study of LTP, we observed that, although the normalized slopes averages in TY have showed always smaller than of Wistar after high frequence stimulus, these values between the two species did not differ statistically. The electroencephalographic and behavioral data were similar between TY and Wistar in the two epilepsy induction models, with exception of the pilocarpine dosage used to elicite status epilepticus in TY, that was lower. The electrophysiological characterization and the obtained results against the epilepsy models are interesting contributions to the knowledge of the nervous system of the TY and reveal the importance to futures works in this species for the acquisition of new knowledges that can to be involved in the genesis of the seizures. / TEDE / BV UNIFESP: Teses e dissertações

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The present research examined the hypothesis that omega-3 polyunsaturated fatty acids would increase seizure threshold in rats in vivo, and reduce neuronal excitability in mouse hippocampal slices. Seizure thresholds were measured in rats using the maximal pentylenetetrazol and electrical stimulation seizure tests following α-linolenic acid (ALA) or docosahexaenoic acid administration. ALA raised seizure threshold in the maximal PTZ seizure test, but this effect probably occurred because ALA displaced DHA from liver to the brain. DHA itself was therefore tested in the PTZ and electrical stimulation seizure tests. Direct administration of DHA by subcutaneous injection raised seizure thresholds in the PTZ seizure test, which models tonic-clonic attacks in humans. Dietary enrichment with DHA raised afterdischarge seizure thresholds in the cortex and amygdala, which model simplex and complex partial seizures in humans, although this effect took some time to occur. In vitro, the application of DHA also reduced the incidence of excitatory sharp waves in mouse hippocampal slices. This effect did not appear to be due to either an increase in GABAergic inhibitory tone, nor to a decrease in glutamatergic drive. The fatty acid composition of phospholipids and unesterified fatty acids were measured in the brain following microwave fixation in order to determine whether the effects of DHA on seizure thresholds were due to its de-esterification from the phospholipid membrane. The assay surprisingly revealed that subcutaneous administration of DHA at a dose that raised seizure threshold, increased unesterified arachidonic acid, but not unesterified DHA concentrations during seizures. The results of these studies support the hypothesis that DHA raises seizure threshold in rats, and reduces neuronal excitability in vitro. The effects of DHA on seizure threshold are possibly mediated by the de-esterification of arachidonic acid, which is known to have effects on the voltage-dependent sodium channel.