Orientador: Elenice Aparecida de Moraes Ferrari / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-06T20:47:31Z (GMT). No. of bitstreams: 1 Siqueira_LuisOtavio_D.pdf: 1043861 bytes, checksum: 39ebfc094cde948f0af9f82213528298 (MD5) Previous issue date: 2006 / Resumo: O comportamento exploratório garante a inspeção, a localização, a identificação e a avaliação precisa de eventos ambientais. A exploração também é abordada como sendo relacionada com a curiosidade do animal direcionada a novos estímulos. Os comportamentos reflexos que precedem as reações exploratórias, caracterizando-se, portanto, como comportamentos pré-exploratórios e também têm importante função adaptativa. Poucos estudos analisaram a influência da hora do dia sobre respostas incondicionadas e condicionadas ao som. Dessa forma, o presente estudo investigou a variação circadiana da amplitude da resposta aversiva ao som em pombos. Experimento I usou um equipamento de registro automatizado da amplitude da resposta acústica, formado por uma câmara experimental, com uma célula de carga acoplada ao piso e interface dos sinais a um microcomputador utilizado para controle, armazenamento e análise dos dados. O Experimento II analisou a sensibilização da resposta em diferentes horários do dia usando: (a) linha de base com apresentação de 5 estímulos acústicos com intervalo entre sons de 3Os; (b) treino, iniciado 5 min após o último som, com dois choques elétricos, intervalo entre choques de 5 min e (c) teste, iniciado 10 min após o último choque, com 40 estímulos acústicos. Os animais dos grupos controles permaneceram na caixa experimental sem estimulação. O Experimento 111 usou intervalos (a) entre o término da linha de base e o início do treino, (b) entre os dois choques e (c) entre o último choque e o teste foram de 1 mino O Experimento IV investigou o condicionamento clássico de traço, utilizando grupos: Experimental, com três associações som-choque; Randômico, com 3 choques e 3 sons casualizados e Controle, expostos à caixa experimental.Em todos os experimentos usou-se fotoperíodo esqueleto com pulsos de luz (15 min) às 6h (ZTOO) e às 18h (ZT12) e sub-grupos em cada horário de teste: ZT02, ZT06, ZT10, ZT14, ZT18 e ZT22. De um modo geral, os dados mostraram que tanto a sensibilização quanto o condicionamento clássico aversivo resultaram em alterações da amplitude da resposta ao som que variaram em função do horário de teste, com amplitudes máximas na transição dia-noite e mínimas na transição noite-dia. O conjunto desses dados indica a modulação do aprendizado aversivo pelo sistema temporizador circadiano / Abstract: Exploratory behavior allows the inspection, localization, identification and the correct evaluation of the environmental events. The exploration also is related with curiosity related to new stimuli. The reflex behaviors that preceed the exploratory reactions are characterized as pre- exploratory and also have an important adaptative function. There are few studies that analyse the influence of time of the day on this response. Thus, the present study investigated the circadian variation of response to aversive sounds in pigeons. Experiment I, used a new equipment especially designed for automatic recording of response of acoustic response amplitude. The system is composed by an experimental chamber, with a load cell under the floor and a computer interface for contrai, storage and data analysis. Experiment 11 analyzed the sensitization of sound response in different times of the day. The experimental protocol used: (a) baseline with 5 acoustic stimulus presentation with inter-stimulus-interval between sounds of the 30s; (b) training, starting 5 min afier the last sound, with apresentation of two electric shocks with interval between electric shocks of the 5 min and (c) test, starting 10 min afier the last shock, with 40 acoustic stimuli. The Control Group animais received no stimulation. Experiment 11I, used 1 min time intervals between (a) the end of the baseline and the trainning, (b) the two electric shocks and (c) the last electric shock and test. Experiment IV investigated the trace fear conditioning, using 2 groups: Experimental, with 3 sound shock associations; Random, with 3 shocks and 3 sounds in the random arder and Contrai, only exposed to experimental chamber. Ali the experiments used a skeleton photoperiod with 2 light pulses (15 min) to 6h (ZTOO) and 18h (ZT12) and sub-groups for each time of test: ZT02, ZT06, ZT10, ZT14, ZT18 and ZT22. In a general, the data showed that both sensitization and classical fear conditioning resulted in increases of the amplitude of sound responses with variations depending on the time of the test. Taken together, the data indicated a modulation of aversive leaming by the circadian pacemaker / Doutorado / Fisiologia / Doutor em Biologia Funcional e Molecular

En permettant aux animaux de faire face à des environnements variables, l'apprentissage et la mémoire contribuent à l'optimisation de leur fitness, en leur permettant d'extraire et d'utiliser des informations, de façon à réduire l'incertitude associée à des environnements imprévisibles. Parmi les insectes, la drosophile et l'abeille domestique sont considérés comme des modèles classiques pour l'étude de l'apprentissage et de la mémoire. Les travaux réalisés sur ces derniers ont apporté une quantité considérable d'informations concernant les bases génétiques, neurobiologiques et moléculaires de ces processus, et ont permis de rendre compte du niveau de complexité des capacités cognitives des insectes. Cette somme de connaissances fondamentales acquises chez ces insectes contraste étonnamment avec le faible niveau de connaissance concernant la cognition des espèces impliquées dans des problématiques qui touchent la santé humaine et animale. Pourtant, il est largement admis que l'étude détaillée des capacités cognitives des insectes vecteurs de maladies constitue un aspect prioritaire pour la compréhension de leurs adaptations à la vie hématophage, de leur importance vectorielle, ainsi que pour le développement de nouveaux outils pour leur contrôle. Les travaux réalisés à ce jour chez les vecteurs, principalement chez les moustiques, ont été menés dans des contextes naturels ou peu contrôlés et ne proposent donc pas de démonstration formelle d'apprentissage. Le principal objectif de ce travail de thèse est de proposer un cadre expérimental contrôlé permettant de mettre en évidence et caractériser les capacités d'apprentissage chez la punaise hématophage \textit{Rhodnius prolixus}. \`A la différence des moustiques, les caractéristiques biologiques de cette punaise hématophage, responsable de la transmission de la Maladie de Chagas en Amérique Latine, permettent l'adaptation de protocoles expérimentaux largement validés chez les drosophiles et l'abeille domestique. Nos résultats montrent dans un premier temps que ces insectes sont capables d'apprendre et d'associer la présentation d'une même odeur dite neutre (l'acide lactique), c'est-à-dire qui ne provoque ni attraction ni répulsion lorsqu'elle est présentée seule, avec soit la possibilité d'obtenir une récompense (un repas sanguin, conditionnement appétitif), soit avec la possibilité de recevoir une punition (un choc mécanique, conditionnement aversif). Nous avons également montré que l'apprentissage et la mémoire sont également impliqués dans le choix des hôtes. Les insectes ont en effet associé la présentation d'un choc mécanique avec le complexe d'odeur d'hôtes naturels, biaisant leur préférence lors d'un test de choix réalisé après l'entraînement. Dans un second temps, nous avons adapté à notre modèle d'étude le paradigme de conditionnement de la réponse d'extension du proboscis, développé chez les modèles classiques, ce qui a permis la caractérisation des capacités d'apprentissage, de la durée de rétention à la régulation par des horloges circadiennes. Ces travaux proposent également un paradigme expérimental, reproductible et efficace permettant d'analyser les mécanismes fins qui sous-tendent les processus d'apprentissage et de mémorisation. Dans son ensemble, cette étude apporte la première preuve expérimentale de la capacité d'apprentissage d'insectes vecteurs de la maladie de Chagas et propose des outils expérimentaux et méthodologiques permettant d'améliorer la compréhension des processus associés chez les insectes hématophages en général. Les résultats sont également discutés dans le contexte de la sélection d'hôte et de la transmission des parasites. / Learning and memory contribute to animals' fitness by allowing them adapting to variable environments. Thses two processes make them able to extract and use information from their environment in order to reduce the uncertainty associated with unpredictible environments. Among insects, fruit flies and honeybees are considered as classical models for the study of learning and memory. The amount of work that has been done on these models provide a considerable amount of information regarding the genetic, neurobiological and molecular basis of these processes and revealed the complexity of insects' cognitive abilities. All this knowledge acquired in model species, contrasts surprisingly with the lack of knowledge available regarding insect species that are involved in animal and human diseases transmission. Yet, it has been aknowledge that the detailed study of vectors cognitive abilities would allow the understanding of their adaptation to haematophagy, of their vectorial importance and provide new tools for diseases control. Up to date, studies focusing on disaese vectors, mainly in mosquitoes, were conducted in natural or not completely controled contexts and thus no clear demonstration of learning and memory is availaible.The main goal of this work was to provide a controled experimental context allowing the strudy of learning abilities in the haematophagous bug \textit{Rhodnius prolixus}. Our results show that these insectes are able to learn to associate the delivery of a same neutral odour either with the possibility to obtain a reward (blood-meal, appetitive conditioning) or with the possibility to receive a punishment (mechanical shock, aversive conditioning). We also showed that learning ans memory are involved in host selection processes. In a second part, we adapted to our biological model the paradigm of proboscis extension response conditioning, which allowed us to analyse and characterize its learning abilities. The maximal retention duration as well as the modulation of learning abilities by circadian clocks were evinced. Taken as a whole, this work provides the first experimental demonstration of learning abilities in Chagas disease vectors and provides experimental and methodological tools; These latters should allow improving the understanding of the mechanisms that are underlying cearning abilities of haematophagous insects in general. Results are also discussed in the context of host selection and parasite transmission.