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Este trabalho de mestrado teve como objetivo o desenvolvimento de uma metodologia analítica focada no preparo de amostra protéica. O objeto de estudo foram os fragmentos solúveis das isoformas da proteína precursora do amilóide (APPs) presentes no plasma rico em plaquetas. As APPs têm sido amplamente estudadas em diversos grupos de pesquisa no Brasil e em outros no mundo como possíveis biomarcadores para a doença de Alzheimer. O preparo de amostra é a etapa fundamental que influencia significativamente nos resultados seguintes, especialmente quando se trata de amostras protéicas que exigem maiores cuidados. Para a avaliação dos melhores preparos de amostra para as APPs, foi utilizado SDS-PAGE e eletrotransferências de proteínas por Western Blotting. A eficiência dos preparos foi avaliada baseando-se nos resultados de revelação com anticorpos específicos para APP e medidas de densitometria de bandas. Após a escolha do melhor preparo de amostra utilizando SDS-PAGE e Western Blotting, as isoformas da APP foram separadas por eletroforese bidimensional (2DE). Durante a etapa de preparo de amostra, os resultados inesperados de massa molecular, o que indicou possível biodegradação das APPs. A identificação da fonte de interferência foi realizada estudando as variáveis dos preparos de amostra. Com isso foi possível determinar a fonte de interferência, mas uma avaliação mais detalhada das isoformas (como utilização de espectrometria de massas) não foi possível. / The goal of this Master\'s work was to develop an analytical methodology focused on protein sample preparation. The analyte studied were soluble amyloid precursor protein isoforms (APPs) which has been studied in many groups in Brazil and around the world as a possible biomarker for Alzheimer\'s disease. Sample preparation is a crucial step that influence significantly on next results, especially about biological samples which require more attention. For the best sample preparation for APPs, was used SDS-PAGE and protein electrotransference by Western Blotting techniques. The efficiency of the sample preparations was evaluated based on specific antibody reactions and densitometry measures of these interactions. After that, the APP isoforms were analyzed by two dimensional electrophoresis (2DE). During the sample preparation, were obtained unexpected molecular mass results, which indicated some APPs biodegradation. For the determination of the interference source, the variants steps of the sample preparation were analyzed. The sample preparation interference source was identified, but a more detailed study of the isoforms (by mass spectrometry) was not possible as well as the analysis of the identity of the possible fragmented isoforms.

A doença de Alzheimer é uma afecção crônica degenerativa que não possui tratamento até o momento. O uso de alimentos funcionais como a romã na prevenção e/ou tratamento de doenças neurodegenerativas têm sido amplamente pesquisados. Diante disto, o presente trabalho teve como objetivo avaliar a quantidade de compostos bioativos (antocianinas, compostos fenólicos e flavonoides), atividade anticolinesterásica e a capacidade antioxidante in vitro e on line dos extratos de polpa e casca de romã e posteriormente estudar o possível efeito neuroprotetor de micropartículas obtidas a partir do extrato da casca de romã em um animal submetido a infusão crônica de peptídeo β-amilóide. O extrato de casca de romã apresentou maior teor de antocianinas, compostos fenólicos, flavonoides e atividade antioxidante in vitro e on line do que o extrato da polpa. Na análise de compostos não voláteis pela técnica de GC-MS foram identificados 38 compostos no extrato da casca e 37 da polpa de romã, sendo o ácido gálico a principal substância detectada. Foram encontrados no total 13 compostos no extrato de casca e 8 no extrato de polpa de romã que apresentaram atividade antioxidante pelo método HPLC-ABTS on line. A punicalagina, epicatequina e ácido gálico foram os compostos determinantes para a atividade antioxidante em ambos os extratos. O extrato da casca de romã apresentou atividade anticolinesterásica superior ao da polpa. Estes resultados, em conjunto, indicaram um possível potencial da casca de romã como um agente neuroprotetor na doença de Alzheimer. Para estudar o possível efeito neuroprotetor do extrato da casca de romã foram utilizados camundongos C57BL/6 cronicamente infundidos com peptídeo βA1-42 e/ou veículo através de mini-bombas osmóticas durante 35 dias e foram avaliados biomarcadores e alterações comportamentais. Micropartículas de extrato de casca de romã, produzidas em spray dryer, foram diluídas em água e administradas na dose de 800 mg de casca de romã/kg de animal/dia. A memória espacial foi avaliada em labirinto de Barnes e uma redução no número de erros para encontrar a caixa de escape foi verificada nos animais tratados com micropartículas de casca de romã e nos animais do grupo controle, mas não nos animais do grupo βA. A atividade da acetilcolinesterase, neurotrofina BDNF, TNF-α e a enzima SOD foram avaliadas no hipocampo, córtex e soro dos animais. A peroxidação lipídica foi avaliada no fígado dos animais. Como a casca de romã não é comumente consumida foram dosados marcadores de dano isquêmico hepático. O consumo de micropartículas de casca de romã promoveu uma redução do acumulo de placas amiloides, aumento da expressão de neurotrofinas, redução da atividade da enzima acetilcolinesterase, redução da peroxidação lipídica e da citocina pró-inflamatória TNF-α em animais infundidos com peptídeo β-amilóide. O consumo das microcáspulas de casca de romã não acarretou nenhum tipo de lesão hepática. No geral, verificou-se que os compostos presentes na casca de romã podem apresentar um efeito neuroprotetor em animais submetidos a infusão crônica de peptídeo β-amilóide. / Alzheimer\'s disease is a chronic and degenerative condition that have no treatment until now. The research of functional foods such as pomegranate for the prevention and/or treatment of many conditions, including neurodegenerative diseases, is increasing year after year. The amount of bioactive compounds (anthocyanins, phenolic compounds and flavonoids), acetylcholinesterase activity and antioxidant capacity in vitro and on line of pomegranate peel and pulp extracts were evaluated. Pomegranate peel extract has higher content of anthocyanins, phenolic compounds, flavonoids and antioxidant activity in vitro and on line than pulp. The analyses of the profile of non-volatile compounds identified 38 compounds in the peel and 37 in the pulp. The gallic acid was main compound detected. Pomegranate peel showed 13 compounds with antioxidant activity by the HPLC-ABTS method online and pulp showed eight compounds. Punicalagin, Gallic acid and epicatechin were determinants for the antioxidant capacity of the aqueous-alcoholic extract of pomegranate. Pomegranate peel extract had greater anticholinesterase activity than pulp. These results together indicated a possible potential of pomegranate peel as a neuroprotective agent in Alzheimer\'s disease. This research had as objective to study the possible neuroprotective effect of pomegranate peel on an animal model of the Alzheimer\'s disease. For that purpose, mice model of Alzheimer\'s disease were used and biomarkers and behavioral changes were evaluated. C57BL/6 mice were chronically infused with βA1-42 peptide and/or vehicle by mini - osmotic pumps during 35 days. Microparticles of pomegranate peel extract, produced by spray drying, were diluted in water and administered at a dose of 800 mg of pomegranate peel/ kg animal/day. The spatial memory was evaluated in the Barnes maze and a reduction of the errors to find the scape box was verified in animals treated with the PPE, as observed in the Control group, but not in th Aβ group. The activity of acetylcholinesterase, neurotrophin BDNF, TNF-α and SOD were measured in the hippocampus, cortex and serum. Lipid peroxidation was evaluated in the liver. As the pomegranate peel is not commonly consumed, biomarkers of liver ischemic damage were measured. Pomegranate peel consumption promoted a reduction of amyloid plaques, increasing neurotrophin expression, reduction in a AChE activity, reduced lipid peroxidation and reduced TNF-α in animal models of Alzheimer\'s disease. The consumption of pomegranate peel did not cause liver injury. In general, pomegranate peel showed a neuroprotective effect on animal models of the Alzheimer\'s disease.

Severas altera¸c~oes no metabolismo energ´etico, no consumo de glicose e na sinaliza¸c~ao de insulina cerebral est~ao presentes na doen¸ca de Alzheimer (DA). O modelo animal da DA obtido pela administra¸c~ao intracerebroventricular de estreptozotocina (STZ-icv) em ratos induz um estado de resist^encia `a insulina no c´erebro associado `a disfun¸c~oes colin´ergicas e a d´eficits cognitivos, tornando-o um dos poucos modelos experimentais da forma espor´adica da DA. Este trabalho tem como objetivo caracterizar, neste modelo, as disfun¸c~oes sin´apticas na via hipocampo - c´ortex pr´e-frontal medial (CA1-CPFm) e testar se o tratamento com nicotina ´e capaz de prevenir as disfun¸c~oes sin´apticas e reverter os preju´?zos cognitivos induzido pelo STZ-icv. Para isso, ratos Wistar receberam STZ e foram submetidos a 20 dias de tratamento com nicotina. Dois dias depois, foram realizados nos animais teste de campo aberto e de reconhecimento de objeto. Em seguida os animais foram anestesiados com uretana para que os registros eletrofisiol´ogicos fossem realizados. Um eletrodo foi utilizado para estimular CA1 com pulso pareado e potenciais de campo p´os-sin´apticos (fPSP1) e sua facilita¸c~ao (fPSP2) foram registradas por um eletrodo no CPFm. Ap´os 30 minutos de linha de base, uma estimula¸c~ao em alta frequ^encia foi aplicada para induzir a potencia¸c~ao de longa dura¸c~ao (LTP), seguido de mais quatro horas de registro. Outro grupo experimental foi realizado para avaliar o efeito de longo prazo da STZ-icv e do tratamento com nicotina. Neste grupo, testes comportamentais e eletrofisiol´ogicos foram realizados 60 dias ap´os o fim do tratamento. Independentemente do tempo, os resultados indicam que a STZ produziu uma redu¸c~ao na indu¸c~ao e na manuten¸c~ao da LTP, mas a facilita¸c~ao por pulso pareado (PPF = fPSP2 / fPSP1) mostra que a STZ prejudica a plasticidade pr´e-sin´aptica apenas a curto prazo. O tratamento com nicotina atenua a disfun¸c~ao na LTP induzida pela STZ. Al´em disso, apenas o tratamento de nicotina tamb´em ´e capaz de reduzir a plasticidade pr´e-sin´aptica no grupo controle dois dias ap´os o fim do tratamento. Estes resultados tamb´em est~ao associados com os dados comportamentais, uma vez que a nicotina reverteu os d´eficits de mem´oria de reconhecimento nos animais STZ mas manteve o comportamento explorat´orio reduzido. Sugerimos com isso que o sistema colin´ergico, que desempenha um papel importante em fun¸c~oes cognitivas e na LTP, est´a afetado nos animais injetados com STZ e o tratamento cr^onico com nicotina consegue reduzir os danos na plasticidade sin´aptica e comportamentais, induzidos pela STZ. / Severe abnormalities in brain glucose/energy metabolism and insulin signaling have been documented to play an important role in early stage of alzheimer disease (AD) pathology. Intracerebroventricular administration (icv) of streptozotocin (STZ) in rats can induce an insulin-resistant brain state associated with cholinergic dysfunctions and memory impairments, which make it a suitable experimental model of the sporadic form of AD. The present work aimed to extend the characterization of this model by probing synaptic plasticity dysfunctions in the medial prefrontal cortex (mPFC)- hippocampal (CA1) pathway and test if nicotine can prevent synaptic dysfunction and revert cognitive impairment induced by icv STZ. Here, Wistar rats received bilateral microinjection of STZ and were submitted to 20 days of nicotine treatment. After 2 days of withdrawing the subjects were submitted to open field and object recognition tests. After that, animals were anesthetized with urethane for electrophysiological tests. A twisted bipolar electrode was used to stimulate posterior-dorsal hippocampus (CA1/subiculum) with paired-pulse. Basal field post-synaptic potentials (fPSP1) and facilitated responses (fPSP2) were recorded by a monopolar electrode in the medial mPFC. After 30min of baseline, high frequency stimulation was applied to induce long-term potentiation (LTP) and additional four hours of electrophysiological recordings was performed. Another experimental group was performed to evaluate the long term effect of both icv STZ and nicotine treatment. In this group behavioral and electrophysiological tests were performed with 60 days after chronic treatment. Independently of time, our results indicate that STZ produced a significant decrease in the induction and maintenance of LTP, but paired pulse facilitation (PPF = fPSP2/fPSP1) shows that only the short-term pre-synaptic plasticity was impaired after STZ injection. The nicotine treatment attenuates the STZ-induced LTP dysfunction in the CA1-mPFC pathway. However, just the nicotine treatment (in control group) can reduce pre-synaptic plasticity two days after chronic treatment. These results are also associated with behavioral data, since nicotine treatment reversed the deficits in recognition memory of STZ animals but maintained the reduced exploratory behavior. We suggest that the brain cholinergic system, which plays a role in cognition function and LTP, is affected in STZ injected animals and chronic treatment with nicotine can attenuate the STZ-induced synaptic plasticity and behavioral dysfunctions.

Este trabalho de mestrado teve como objetivo o desenvolvimento de uma metodologia analítica focada no preparo de amostra protéica. O objeto de estudo foram os fragmentos solúveis das isoformas da proteína precursora do amilóide (APPs) presentes no plasma rico em plaquetas. As APPs têm sido amplamente estudadas em diversos grupos de pesquisa no Brasil e em outros no mundo como possíveis biomarcadores para a doença de Alzheimer. O preparo de amostra é a etapa fundamental que influencia significativamente nos resultados seguintes, especialmente quando se trata de amostras protéicas que exigem maiores cuidados. Para a avaliação dos melhores preparos de amostra para as APPs, foi utilizado SDS-PAGE e eletrotransferências de proteínas por Western Blotting. A eficiência dos preparos foi avaliada baseando-se nos resultados de revelação com anticorpos específicos para APP e medidas de densitometria de bandas. Após a escolha do melhor preparo de amostra utilizando SDS-PAGE e Western Blotting, as isoformas da APP foram separadas por eletroforese bidimensional (2DE). Durante a etapa de preparo de amostra, os resultados inesperados de massa molecular, o que indicou possível biodegradação das APPs. A identificação da fonte de interferência foi realizada estudando as variáveis dos preparos de amostra. Com isso foi possível determinar a fonte de interferência, mas uma avaliação mais detalhada das isoformas (como utilização de espectrometria de massas) não foi possível. / The goal of this Master\'s work was to develop an analytical methodology focused on protein sample preparation. The analyte studied were soluble amyloid precursor protein isoforms (APPs) which has been studied in many groups in Brazil and around the world as a possible biomarker for Alzheimer\'s disease. Sample preparation is a crucial step that influence significantly on next results, especially about biological samples which require more attention. For the best sample preparation for APPs, was used SDS-PAGE and protein electrotransference by Western Blotting techniques. The efficiency of the sample preparations was evaluated based on specific antibody reactions and densitometry measures of these interactions. After that, the APP isoforms were analyzed by two dimensional electrophoresis (2DE). During the sample preparation, were obtained unexpected molecular mass results, which indicated some APPs biodegradation. For the determination of the interference source, the variants steps of the sample preparation were analyzed. The sample preparation interference source was identified, but a more detailed study of the isoforms (by mass spectrometry) was not possible as well as the analysis of the identity of the possible fragmented isoforms.

L’objectif de cette thèse est d’étudier, sous divers aspects, le processus de formation d’amyloïde à partir de la polymérisation de protéines. Ces phénomènes, aussi bien in vitro que in vivo, posent des questions de modélisation mathématique. Il s’agit ensuite de conduire une analyse des modèles obtenus. Dans la première partie nous présentons des travaux effectués en collaboration avec une équipe de biologistes. Deux modèles sont introduits, basés sur la théorie en vigueur du phénomène Prions, que nous ajustons aux conditions expérimentales. Ces modèles nous permettent d’analyser les données obtenues à partir d’expériences conduites en laboratoire. Cependant celles-ci soulèvent certains phénomènes encore inexpliqués par la théorie actuelle. Nous proposons donc un autre modèle qui corrobore les données et donne une nouvelle approche de la formation d’amyloïde dans le cas du Prion. Nous terminons cette partie par l’analyse mathématique de ce système compose d’une infinité d’équations différentielles. Ce dernier consiste en un couplage entre un système de type Becker-Doring et un système de polymérisation-fragmentation discrète. La seconde partie s’attache à l’analyse d’un nouveau modèle pour la polymérisation de protéines dont la fragmentation est sujette aux variations du fluide environnant. L’idée est de décrire au plus près les conditions expérimentales mais aussi d’introduire de nouvelles quantités macroscopiques mesurables pour l’étude de la polymérisation. Le premier chapitre de cette partie présente une description stochastique du problème. On y établit les équations du mouvement des polymères et des monomères (de type Langevin) ainsi que le formalisme pour l’étude du problème limite en grand nombre. Le deuxième chapitre pose le cadre fonctionnel et l’existence de solutions pour l équation de Fokker-Planck- Smoluchowski décrivant la densité de configuration des polymères, elle-même couplée a une équation de diffusion pour les monomères. Le dernier chapitre propose une méthode numérique pour traiter ce problème. On s’intéresse dans la dernière partie à la modélisation de la maladie d’Alzheimer. On construit un modèle qui décrit d’une part la formation de plaque amyloïde in vivo, et d’autre part les interactions entre les oligomères d’Aβet la protéine prion qui induiraient la perte de mémoire. On mène l’analyse mathématique de ce modèle dans un cas particulier puis dans un cas plus général ou le taux de polymérisation est une loi de puissance / The aim of this thesis is to study, under several aspects, the formation of amyloids from proteins polymerization. The mathematical modelling of these phenomena in the case of in vitro or in vivo polymerisation remains questioned. We then propose here several models, which are also investigated from theoritical and numerical point of view. In the first part we present works done in collaboration with biologists. We propose two models based on the current theory on Prion phenomena that are designed for specific experimental conditions. These models allow us to analyse the experimental data obtained in laboratory and raise phenomena that remain unexplained by the theory. Then, from these results and biophysical considerations, we introduce a model which corroborates with data and provides a new approach on the amyloid formation in the particular case of Prion. This part is ended by the mathematical analysis of the model consisting of an infinite set of differentials equations. The system analysed is a Becker-Doring system coupled to a discrete growth-fragmentation system. The second part is dedicated to the analysis of a new model for polymerization of proteins with fragmentation subject to the surrounding variations of the fluid. Thus, we propose a model which is close to the experimental conditions and introduce new measurable macroscopic quantities to study the polymerization. The first introductory chapter states the stochastic description of the problem. We give the equations of motion for each polymers and monomers as well as a general formalism to study the limit in large number. Next, we give the mathematical framework and prove the existence of solutions to the Fokker-Planck-Smoluchowski equation for the configurational density of polymers coupled to the diffusion equation for monomers. The last chapter provides a numerical method adapted to this problem with numerical simulations In the last part, we are interested in modelling Alzheimer’s disease. We introduce a model that describes the formation of amyloids plaques in the brain and the interactions between Aβ-oligomers and Prion proteins which might be responsible of the memory impairment. We carry out the mathematical analysis of the model. Namely, for a constant polymerization rate, we provide existence and uniqueness together with stability of the equilibrium. Finally we study the existence in a more general and biological relevant case, that is when the polymerization depends on the size of the amyloid

La maladie d’Alzheimer et les pathologies apparentées, en évoluant génèrent une altération de l’état général, et une dépendance générale. Le patient, dont les capacités d’adaptation sont altérées par la maladie risque, à tous les stades de la maladie de réagir par des troubles du comportement. Ce sont ces troubles qui le conduisent généralement à entrer en institution spécialisée. L’institution doit alors prendre en charge la dépendance physique et psychique du patient. Elle doit, mettre en place une prévention des complications de la maladie. Avec parmi celles-ci les manifestations comportementales perturbantes. Cela nécessite à la fois des équipes spécialement formées et des établissements ou des unités de soins adaptés à cette prise en charge spécifique. Cette thèse tente de répondre à une question simple : sur quoi s’appuient au quotidien les soignants pour parler d’ « amélioration » ou d’ « aggravation », sans recourir à l’évaluation avec des tests en condition standard ? Et cela, en sachant que les conditions standard qui sont valides pour la recherche, sont des conditions artificielles qui ne permettent pas de caractériser toutes les potentialités des malades. Le présent travail, effectué dans une unité spécialisée, a pour objectif de tenter de décoder ce qui détermine subjectivement le jugement des soignants à partir d’une analyse de leur discours dans le cadre d’entretiens semi-directifs. Ce discours a successivement fait l’objet d’une analyse informatique au moyen du logiciel « Alceste » puis d’une analyse thématique de contenu, dite : « manuelle ». Les résultats de ces deux analyses sont exposés en miroir de ceux des tests « standards » (MMS ; GDS ; NPI/ES ZARIT). Tests qui ont permis de distinguer quels patients, d’un point de vue classique, avaient été stables, mais aussi s’étaient améliorés ou aggravés en six mois. La seconde analyse a permis de mettre très explicitement en évidence que les préoccupations dominantes des soignants portaient sur des éléments descriptifs. C’est-à-dire sur les tâches qu’ils ont à faire matériellement. Il apparait que pour les soignants la notion d’amélioration ou d’aggravation du malade est en résumé associée à la charge de travail que son état implique. Cela, avec entre autres des références aux conduites gênantes et aux problèmes médicaux. On observe de plus que le patient est peu considéré comme un sujet auquel s’identifier. Des facteurs psychologiques peuvent cependant être envisagés par les soignants lorsqu’il s’agit d’expliquer certaines conduites perturbantes. / The Alzheimer's disease and the related pathologies, by evolving, generate a change of the the overall condition of the patient, and a general dependence. The patient, whose capacities of adaptation are affected by the disease risks, in all the stages of the disease, to react by behavior disorders. It is these disorders which lead him generally to enter specialized institution.The institution then has to take care of the physical and psychic dependence of the patient. She owes to set up a prevention of the complications of the disease. With among these the disturbing behavioral manifestations. It requires at the same time specially trained teams and establishments or units of care adapted to this specific management.This thesis tries to answer a simple question: on what base themselves, on a daily basis, the nursing to speak about "improvement" or about "worsening", without using of the evaluation with tests in standard condition?And these, by knowing that the standard conditions which are valid for the search, are artificial conditions which do not allow to characterize all the potentialities of the disease.The present work, made in a specialized unit, aims to decode what determines subjectively the assessment of the nursing from an analysis of their discourse within semi-directive discussions.This discourse was successively the object of an IT analysis by the software "Alceste", then a thematic analysis of contents, called: "manual".The results of these two analyses are mirrored with those of the "standard" tests (MMS; GDS; NPI/ES ZARIT). Tests which allowed to distinguish which patients, from a classic point of view, had been stable, but also had improved or deteriorated in six months.The second analysis allowed to highlight very explicitly that the dominant concerns of the nursing was about descriptive elements. That is on the tasks which they have to do materially.It seems that for the nursing the notion of improvement or worsening of the patient is, in summary, correlated to the workload it involves. Due, among others, to the annoying behavior and to the medical problems…We observe, furthermore, that the patient is a bit considered as a subject in which become identified. Psychological factors can be however envisaged by the nursing when it is a question of explaining certain disturbing behavior.

Severas altera¸c~oes no metabolismo energ´etico, no consumo de glicose e na sinaliza¸c~ao de insulina cerebral est~ao presentes na doen¸ca de Alzheimer (DA). O modelo animal da DA obtido pela administra¸c~ao intracerebroventricular de estreptozotocina (STZ-icv) em ratos induz um estado de resist^encia `a insulina no c´erebro associado `a disfun¸c~oes colin´ergicas e a d´eficits cognitivos, tornando-o um dos poucos modelos experimentais da forma espor´adica da DA. Este trabalho tem como objetivo caracterizar, neste modelo, as disfun¸c~oes sin´apticas na via hipocampo - c´ortex pr´e-frontal medial (CA1-CPFm) e testar se o tratamento com nicotina ´e capaz de prevenir as disfun¸c~oes sin´apticas e reverter os preju´?zos cognitivos induzido pelo STZ-icv. Para isso, ratos Wistar receberam STZ e foram submetidos a 20 dias de tratamento com nicotina. Dois dias depois, foram realizados nos animais teste de campo aberto e de reconhecimento de objeto. Em seguida os animais foram anestesiados com uretana para que os registros eletrofisiol´ogicos fossem realizados. Um eletrodo foi utilizado para estimular CA1 com pulso pareado e potenciais de campo p´os-sin´apticos (fPSP1) e sua facilita¸c~ao (fPSP2) foram registradas por um eletrodo no CPFm. Ap´os 30 minutos de linha de base, uma estimula¸c~ao em alta frequ^encia foi aplicada para induzir a potencia¸c~ao de longa dura¸c~ao (LTP), seguido de mais quatro horas de registro. Outro grupo experimental foi realizado para avaliar o efeito de longo prazo da STZ-icv e do tratamento com nicotina. Neste grupo, testes comportamentais e eletrofisiol´ogicos foram realizados 60 dias ap´os o fim do tratamento. Independentemente do tempo, os resultados indicam que a STZ produziu uma redu¸c~ao na indu¸c~ao e na manuten¸c~ao da LTP, mas a facilita¸c~ao por pulso pareado (PPF = fPSP2 / fPSP1) mostra que a STZ prejudica a plasticidade pr´e-sin´aptica apenas a curto prazo. O tratamento com nicotina atenua a disfun¸c~ao na LTP induzida pela STZ. Al´em disso, apenas o tratamento de nicotina tamb´em ´e capaz de reduzir a plasticidade pr´e-sin´aptica no grupo controle dois dias ap´os o fim do tratamento. Estes resultados tamb´em est~ao associados com os dados comportamentais, uma vez que a nicotina reverteu os d´eficits de mem´oria de reconhecimento nos animais STZ mas manteve o comportamento explorat´orio reduzido. Sugerimos com isso que o sistema colin´ergico, que desempenha um papel importante em fun¸c~oes cognitivas e na LTP, est´a afetado nos animais injetados com STZ e o tratamento cr^onico com nicotina consegue reduzir os danos na plasticidade sin´aptica e comportamentais, induzidos pela STZ. / Severe abnormalities in brain glucose/energy metabolism and insulin signaling have been documented to play an important role in early stage of alzheimer disease (AD) pathology. Intracerebroventricular administration (icv) of streptozotocin (STZ) in rats can induce an insulin-resistant brain state associated with cholinergic dysfunctions and memory impairments, which make it a suitable experimental model of the sporadic form of AD. The present work aimed to extend the characterization of this model by probing synaptic plasticity dysfunctions in the medial prefrontal cortex (mPFC)- hippocampal (CA1) pathway and test if nicotine can prevent synaptic dysfunction and revert cognitive impairment induced by icv STZ. Here, Wistar rats received bilateral microinjection of STZ and were submitted to 20 days of nicotine treatment. After 2 days of withdrawing the subjects were submitted to open field and object recognition tests. After that, animals were anesthetized with urethane for electrophysiological tests. A twisted bipolar electrode was used to stimulate posterior-dorsal hippocampus (CA1/subiculum) with paired-pulse. Basal field post-synaptic potentials (fPSP1) and facilitated responses (fPSP2) were recorded by a monopolar electrode in the medial mPFC. After 30min of baseline, high frequency stimulation was applied to induce long-term potentiation (LTP) and additional four hours of electrophysiological recordings was performed. Another experimental group was performed to evaluate the long term effect of both icv STZ and nicotine treatment. In this group behavioral and electrophysiological tests were performed with 60 days after chronic treatment. Independently of time, our results indicate that STZ produced a significant decrease in the induction and maintenance of LTP, but paired pulse facilitation (PPF = fPSP2/fPSP1) shows that only the short-term pre-synaptic plasticity was impaired after STZ injection. The nicotine treatment attenuates the STZ-induced LTP dysfunction in the CA1-mPFC pathway. However, just the nicotine treatment (in control group) can reduce pre-synaptic plasticity two days after chronic treatment. These results are also associated with behavioral data, since nicotine treatment reversed the deficits in recognition memory of STZ animals but maintained the reduced exploratory behavior. We suggest that the brain cholinergic system, which plays a role in cognition function and LTP, is affected in STZ injected animals and chronic treatment with nicotine can attenuate the STZ-induced synaptic plasticity and behavioral dysfunctions.

Diss. (sammanfattning) Stockholm : Karol. inst., 2004. / Härtill 4 uppsatser.

Thèse (M.A.)--Université de Sherbrooke (Canada), 2006. / Titre de l'écran-titre (visionné le 8 mai 2008). In ProQuest dissertations and theses. Publié aussi en version papier.