Análise imunoistoquímica do processo oponente da regulação afetiva em resposta ao uso crônico de álcool
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2018-10-25
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Dissertação de mestrado
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Objetivo: Os neurotransmissores dopamina e CRF têm sido associados, respectivamente, às vias responsáveis pelo efeito reforçador positivo e negativo do álcool. Este estudo teve como objetivos: a) analisar os comportamentos motor e associado à ansiedade de ratos submetidos a um protocolo de ingestão crônica de álcool por quatro semanas; b) analisar, nos cérebros dos mesmos animais, quanto à imunorreatividade aos neurotransmissores Dopamina e CRF. Material e Métodos: Foram utilizados 24 ratos Wistar provenientes do CEDEME/UNIFESP, submetidos ao protocolo de ingestão voluntária de solução alcoolizada a 20% (paradigma das duas garrafas) de caráter intermitente (álcool oferecido durante as segundas, quartas e sextasfeiras) durante quatro semanas. O teste do Campo Aberto foi utilizado para análise motora e o teste do Labirinto em Cruz Elevado para análise dos comportamentos associados à ansiedade. Após perfusão e preparação histológica do tecido cerebral contendo áreas de interesse do sistema dopaminérgico e CRFérgico, foi realizada a análise de imunorreatividade aos neurotransmissores dos respectivos sistemas através da técnica de imunoperoxidase. Resultados: A análise do Campo Aberto permitiu classificar os animais que consumiram álcool em relação à atividade motora (alta atividade e baixa atividade). Os resultados obtidos no teste do Labirinto em Cruz Elevado mostraram que os animais de alta atividade produziram menos bolos fecais indicando efeito ansiolítico do álcool sobre esses animais. A análise imunoistoquímica demonstrou que animais que ingeriram álcool cronicamente apresentaram menor imunorreatividade à dopamina quando comparados ao controle. Conclusões: Por meio do presente estudo podese concluir que animais que consumiram solução alcoolizada de forma crônica e intermitente puderam ser classificados em animais de alta atividade e de baixa atividade motora, e os considerados de alta atividade foram aqueles que consumiram significativamente mais álcool. Na dose utilizada, não foi observado efeito ansiolítico do etanol nos testes comportamentais, assim como nenhum efeito sobre o sistema cerebral de estresse. No entanto, foi observado efeito inibitório do consumo crônico de álcool sobre o sistema cerebral de recompensa dopaminérgico, independente do perfil de alta ou de baixa atividade motora.
Objective: The neurotransmitters dopamine and CRF have been associated, respectively, to the neural pathways responsible for the positive and negative reinforcing effect of alcohol. This study had the following objectives: a) to analyze the motor and anxietyassociated behaviors of rats submitted to a protocol of chronic alcohol ingestion for four weeks; b) to analyze, in the brains of the same animals, the immunoreactivity to the neurotransmitters Dopamine and CRF. Material and Methods: Twentyfour Wistar rats from CEDEME/UNIFESP were submitted to a voluntary ingestion protocol of alcoholic solution 20% (twobottle choice paradigm) of an intermittent nature (alcohol was offered on Mondays, Wednesdays, and Fridays) for four weeks. The Open Field test was used for cerebral motor activity analysis and the Elevated Plus Maze test for the analysis of the behaviors associated with anxiety. After perfusion and histological preparation of brain tissue containing areas of interest of the dopaminergic and CRFergic systems, immunoreactivity analysis was performed on the respective systems utilizing the immunoperoxidase technique. Results: The Open Field test analysis allowed the classification of the alcohol consuming animals in high motor activity and low motor activity. The results obtained from the Elevated Plus Maze test showed that the high activity animals produced fewer fecal boli indicating an anxiolytic effect of alcohol on these animals. Immunohistochemical analysis showed that animals that chronically ingested alcohol solution showed lower immunoreactivity to dopamine when compared with the control group. Conclusions: Through the present study, it can be concluded that animals that consumed alcoholic solution in a chronic and intermittent manner could be classified as animals of high or low motor activity and those classified as high motor activity animals were those that consumed significantly more alcohol. No anxiolytic effect of ethanol was observed in the behavioral tests, as well as no effect on the brain stress system. However, we observed an inhibitory effect of chronic alcohol consumption on the dopaminergic reward brain system, regardless of the motor activity classification.
Objective: The neurotransmitters dopamine and CRF have been associated, respectively, to the neural pathways responsible for the positive and negative reinforcing effect of alcohol. This study had the following objectives: a) to analyze the motor and anxietyassociated behaviors of rats submitted to a protocol of chronic alcohol ingestion for four weeks; b) to analyze, in the brains of the same animals, the immunoreactivity to the neurotransmitters Dopamine and CRF. Material and Methods: Twentyfour Wistar rats from CEDEME/UNIFESP were submitted to a voluntary ingestion protocol of alcoholic solution 20% (twobottle choice paradigm) of an intermittent nature (alcohol was offered on Mondays, Wednesdays, and Fridays) for four weeks. The Open Field test was used for cerebral motor activity analysis and the Elevated Plus Maze test for the analysis of the behaviors associated with anxiety. After perfusion and histological preparation of brain tissue containing areas of interest of the dopaminergic and CRFergic systems, immunoreactivity analysis was performed on the respective systems utilizing the immunoperoxidase technique. Results: The Open Field test analysis allowed the classification of the alcohol consuming animals in high motor activity and low motor activity. The results obtained from the Elevated Plus Maze test showed that the high activity animals produced fewer fecal boli indicating an anxiolytic effect of alcohol on these animals. Immunohistochemical analysis showed that animals that chronically ingested alcohol solution showed lower immunoreactivity to dopamine when compared with the control group. Conclusions: Through the present study, it can be concluded that animals that consumed alcoholic solution in a chronic and intermittent manner could be classified as animals of high or low motor activity and those classified as high motor activity animals were those that consumed significantly more alcohol. No anxiolytic effect of ethanol was observed in the behavioral tests, as well as no effect on the brain stress system. However, we observed an inhibitory effect of chronic alcohol consumption on the dopaminergic reward brain system, regardless of the motor activity classification.
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CONTE, RAFAEL Análise imunoistoquímica do processo oponente da regulação afetiva em resposta ao uso crônico de álcool. 2018. 63 f. Dissertação (Mestrado em Neurologia e Neurociências. ) - Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, 2003.