The dramatic feeding-related activities of the Chagas' disease vector, Rhodnius prolixus are under neurohormonal regulation of serotonin and various neuropeptides. One such family of neuropeptides, the insect kinins, possesses diuretic, digestive and myotropic activities in many insects. In R. prolixus, they co-localize with the corticotropin-releasing factor (CRF)-like diuretic hormone (DH) in neurosecretory cell bodies and their abdominal neurohaemal sites. Additionally, kinins are present in endocrine cells of the midgut and are known to stimulate hindgut and midgut contractions. Through the experimentation presented in this dissertation, the cloning and spatial expression of the R. prolixus kinin (Rhopr-kinin) transcript is described. Physiological bioassays demonstrate the myostimulatory effects of selected Rhopr-kinin peptides and also illustrate the augmented responses of hindgut contractions to co-application of Rhopr-kinin and Rhopr-CRF/DH. The irreversible effects of two synthetic kinin analogs on the hindgut relative to the native kinins also exhibit the prospective biotechnological significance of this study.

The dramatic feeding-related activities of the Chagas' disease vector, Rhodnius prolixus are under neurohormonal regulation of serotonin and various neuropeptides. One such family of neuropeptides, the insect kinins, possesses diuretic, digestive and myotropic activities in many insects. In R. prolixus, they co-localize with the corticotropin-releasing factor (CRF)-like diuretic hormone (DH) in neurosecretory cell bodies and their abdominal neurohaemal sites. Additionally, kinins are present in endocrine cells of the midgut and are known to stimulate hindgut and midgut contractions. Through the experimentation presented in this dissertation, the cloning and spatial expression of the R. prolixus kinin (Rhopr-kinin) transcript is described. Physiological bioassays demonstrate the myostimulatory effects of selected Rhopr-kinin peptides and also illustrate the augmented responses of hindgut contractions to co-application of Rhopr-kinin and Rhopr-CRF/DH. The irreversible effects of two synthetic kinin analogs on the hindgut relative to the native kinins also exhibit the prospective biotechnological significance of this study.

CAPA-related peptides belong to a family of neuropeptides localized to the central nervous system that can function in diverse roles in the regulation of water and salt homeostasis in insects. These peptides are known to stimulate fluid secretion by Malpighian tubules (MTs) in Dipteran species, thus serving a diuretic function. In contrast, this thesis demonstrates that members of this family of peptides in Rhodnius prolixus serve an anti-diuretic role and have multiple tissue targets, whereby they oppose the activity of diuretic hormones such as serotonin (5-Hydroxytryptamine hydrochloride; 5-HT). I have identified two genes each encoding three peptides in R. prolixus, suggesting this insect is capable of producing a greater number of CAPA-peptides compared to other insects that contain only a single CAPA gene. Interestingly, while the second peptide encoded in each R. prolixus gene (RhoprCAPA-α2/-β2) inhibits the stimulatory effects of serotonin on tissues such as the anterior midgut and Malpighian tubules, it appears the other CAPA-related and pyrokinin-related peptides do not play a major role in inhibiting the effects of serotonin on these tissues. More specifically, serotonin-stimulated fluid secretion by MTs and fluid absorption by the anterior midgut are reduced by the anti-diuretic peptide, RhoprCAPA-α2. In addition, I have also identified a G protein-coupled receptor which likely mediates the anti-diuretic effect associated with RhoprCAPA-α2 and have functionally characterized this receptor in Chinese hamster ovary cells. Spatial transcript expression analysis in fifth-instars reveals a wide distribution of the receptor in tissues associated with the rapid post-gorging diuresis. Thus, my findings suggest that numerous tissues are regulated by the CAPA peptides in R. prolixus. Gene structure and phylogenetic analyses demonstrate that this receptor is the orthologue of the D. melanogaster capa receptor (CG14575) with homologs in other insects. Taken together, my thesis demonstrates that the RhoprCAPA peptides play an integral role in the coordination and maintenance of anti-diuresis in R. prolixus. This mechanism is necessary following the rapid diuresis associated with blood-feeding by this medically-important insect.

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.

<p>This thesis analyses several factors affecting the control of triatomines in Guatemala. There are three synantropic triatomines in Guatemala, i.e., <i>Rhodnius prolixus</i>, <i>Triatoma dimidiata</i> and <i>T. nitida</i>. Their distibution is mainly at an altitude between 800 and 1500 m a.s.l. <i>R. prolixus</i> and <i>T. nitida</i> have localized but scaterred distibution while <i>T. dimidiata</i> is present in 21 of the 22 departments in the country. Several investigations have shown that <i>R. prolixus</i> could be relatively easily eradicated while <i>T. dimidiata</i> may be more difficult to control, since it is present in domestic, peridomestic and sylvatic environments showing high diversity and a variety of epidemiological characteristics. Based on the incidence of <i>Trypanosma cruzi</i> infection in humans in the distributional areas of the triatomines, <i>R. prolixus</i> appear to be a more competent vector than <i>T. dimidiata</i>. This is despite the fact that these vectors have similar infection rates. Inside houses, <i>R. prolixus</i> and <i>T. dimidiata</i> and in artificial environments, <i>T. ryckmani</i> and <i>T. dimidiata</i>, preferred the northern side of the walls. Therefore, selective application of insecticides should focus on walls and furniture located in the northern part of the house. House improvements reduced the infestation of triatomines, and could be used as a complement to insecticidal spraying. Although <i>T. dimidiata</i> is not an efficient vector its wide distribution, versatility in occupying different habitats and capacity to disperse render this species difficult to control in Central America. Thus, only few months after insecticidal spraying <i>T. dimidiata</i> had reinfested the domestic environments. Morphometic methodology and genetic markers have been developed to differentiate within-species populations of <i>T. dimidiata</i> and <i>T. nitida</i>. Studies on the migration patterns of sylvatic <i>T. dimidiata</i> and <i>T. ryckmani</i> have been performed in order to clarify the colonization patterns. The adults migrate, in particular, in the dry part of the year. This finding may be of help in attempts to control <i>T. dimidiata</i>.</p>

This thesis analyses several factors affecting the control of triatomines in Guatemala. There are three synantropic triatomines in Guatemala, i.e., Rhodnius prolixus, Triatoma dimidiata and T. nitida. Their distibution is mainly at an altitude between 800 and 1500 m a.s.l. R. prolixus and T. nitida have localized but scaterred distibution while T. dimidiata is present in 21 of the 22 departments in the country. Several investigations have shown that R. prolixus could be relatively easily eradicated while T. dimidiata may be more difficult to control, since it is present in domestic, peridomestic and sylvatic environments showing high diversity and a variety of epidemiological characteristics. Based on the incidence of Trypanosma cruzi infection in humans in the distributional areas of the triatomines, R. prolixus appear to be a more competent vector than T. dimidiata. This is despite the fact that these vectors have similar infection rates. Inside houses, R. prolixus and T. dimidiata and in artificial environments, T. ryckmani and T. dimidiata, preferred the northern side of the walls. Therefore, selective application of insecticides should focus on walls and furniture located in the northern part of the house. House improvements reduced the infestation of triatomines, and could be used as a complement to insecticidal spraying. Although T. dimidiata is not an efficient vector its wide distribution, versatility in occupying different habitats and capacity to disperse render this species difficult to control in Central America. Thus, only few months after insecticidal spraying T. dimidiata had reinfested the domestic environments. Morphometic methodology and genetic markers have been developed to differentiate within-species populations of T. dimidiata and T. nitida. Studies on the migration patterns of sylvatic T. dimidiata and T. ryckmani have been performed in order to clarify the colonization patterns. The adults migrate, in particular, in the dry part of the year. This finding may be of help in attempts to control T. dimidiata.

Les insectes sont soumis aux fluctuations thermiques de leur environnement mais disposent d’un panel varié de réponses comportementales, physiologiques et biochimiques pour en minimiser les effets délétères et maintenir leur intégrité physiologique. Ainsi certaines espèces régulent activement leur température interne indépendamment de la température de l’environnement. Si ces insectes peuvent s’affranchir des contraintes thermiques imposées par leur environnement, ceux qui se nourrissent du sang chaud d’hôtes vertébrés endothermes n’ont pas d’autres choix que de se confronter à une situation de stress thermique à chaque prise alimentaire. Le principal objectif de ce travail de thèse est de comprendre comment des insectes hématophages, employant des stratégies alimentaires différentes, gèrent le stress thermique associé au flux massif de chaleur engendré par l’ingestion du repas de sang. Nos résultats montrent que ces insectes ont su s’adapter en développant différentes stratégies de thermorégulation. / Insects are submitted to thermal fluctuations of their environment and have developed a wide ranged panel of behavioral, physiological and biochemical responses, to minimize the subsequent deleterious effects and maintain their physiological integrity. Some species actively regulate their internal temperature independently of the temperature of the environment. If these insects can overcome the constraints imposed by their thermal environment, those that feed on warm-blooded vertebrate hosts have no choice but to confront a situation of thermal stress at each feeding event. The main objective of this work is to understand how bloodsucking insects manage heat stress associated with the massive flow of heat generated by the ingestion of the blood meal. Our results show these insects have developed different strategies of thermoregulation to protect themselves from overheating.