Ce travail de thèse porte sur l’effet du rayonnement UV-B sur la physiologie foliaire et la maturation des baies chez la Vigne (Vitis vinifera L. cv Tempranillo), dans le contexte du changement climatique en cours. Dans ce but, des expériences ont été menées en conditions contrôlées en serre, sur des boutures fructifères. Les plantes ont été exposées à trois doses de rayonnement UV-B biologiquement actifs (0, 5,98 et 9,66 kJ.m-2.jour-1), soit à partir de la nouaison, soit à partir de la véraison, et ce jusqu’à la pleine maturité. Le rayonnement UV-B a été appliqué seul ou en combinaison avec d’autres stress abiotiques (déficit hydrique, taux de CO2 et température élevés). L’impact de ces stress sur l’activité photosynthétique, les contenus en pigments et en composés photoprotecteurs, ainsi que sur les activités des enzymes produisant des composés antioxydants, ont été étudiés. La phénologie, les profils d’accumulation des flavonols et des anthocyanes, ainsi que le contenu en acides aminés des baies ont également été analysés, de même que l’expression des principaux gènes régulateurs et structuraux de la voie de biosynthèse des polyphénols. Les résultats obtenus montrent que les effets des UV-B sur la physiologie foliaire peuvent être découpés en deux phases : une première phase transitoire de diminution de l’activité photosynthétique, suivie d’une phase d’acclimatation due à la production de composés photoprotecteurs (flavonoïdes essentiellement) et à l’activité d’enzyme de détoxification des forme actives de l’oxygène (superoxyde dimutase, catalase, ascorbate peroxidase). Le déficit hydrique, le stress thermique et un taux de CO2 élevé (700ppm, +4ºC) ne modifient pas le processus d’acclimatation aux UV-B ; en revanche on note une interaction positive entre les UV-B d’une part et la tolérance aux températures et au taux de CO2 élevé d’autre part, atténuant les dommages oxydatifs dus à l’induction de sénescence par ces deux derniers facteurs. La maturité des baies est retardée par les UV-B et le déficit hydrique, et ce phénomène et amplifiés lorsque ces deux stress sont appliqués en combinaison ; alors que les hautes températures et un taux de CO2 élevé ont l’effet inverse. Dans ce dernier cas, le UV-B rayonnement atténue les effets du couple température/CO2 élevés. Ces effets sur la phénologie de la Vigne ont pu être reliés à des modifications de l’activité photosynthétique des feuilles. En ce qui concerne la composition des baies, l’augmentation du rayonnement UV-B et le déficit hydrique ont augmenté les concentrations en anthocyanes et en potassium des moûts et diminué leur acidité, ce qui peut s’expliquer en partie par une augmentation du ratio de masse pellicule/pulpe. L’augmentation des teneurs en anthocyanes et flavonols des pellicules observée en réponse aux UV-B a pu être reliée à l’induction de gènes structuraux (CHS, F3’H, FLS, UFGT and GST) et régulateurs (MYBF1 et MYBA1) de la voie de biosynthèse des flavonoïdes. Ces changements quantitatifs étaient toujours associés à des changements qualitatifs, avec une augmentation de la part relative des flavonols mono- et disubstitués, en raison d’une compétition de la flavonol synthase avec les F3’ et F3’5’ hydroxylases pour les mêmes substrats. Une interaction notable a été observée entre le rayonnement UV-B et le déficit hydrique, sur les profils d’hydroxylation des flavonols, ce qui s’explique par le fait que ces deux facteurs agissent sur deux étapes distinctes de la voie de biosynthèse. / The aim of the thesis was to assess the effect of UV-B radiation on grapevine Vitis viniferacv. Tempranillo leaf physiology and grape berry composition, framed within the climatechange scenario. Experiments were conducted under glasshouse controlled conditions withfruit-bearing cuttings. Plants were exposed to three UV-B biologically effective doses (0,5.98, 9.66 kJ m-2 d-1) either from fruit set or veraison to maturity. The combined effects of UVand water deficit, as well as, UV-B and elevated CO2-temperature (700ppm, +4ºC), appliedfrom fruit set to maturity were also tested. Gas exchange, Chlorophyll a fluorescence, lipidperoxidation, antioxidant enzyme activity, UV-B absorbing compound levels and chlorophylland carotenoid concentration were determined in leaves. Berry development was assessedquantitatively (e.g. elapsed time to reach phenological stages). Amino acid, anthocyanin andflavonol concentrations and profiles were analyzed in berries, as well as, transcript profilingof regulatory and structural genes involved in flavonoid biosynthesis.The results show that initial down-regulation of photosynthesis was followed by anacclimation, mediated by the accumulation of UV-B absorbing compounds and antioxidantresponse elicitation (flavonoids and antioxidant enzymes). Water deficit and elevated CO2-temperature did not alter UV-B acclimation process, however, UV-B did led to certain degreeof cross-tolerance to elevated CO2-temperature, avoiding the senescence-induced oxidativedamage. Berry technological maturity (ca. 22ºBrix) was delayed by UV-B exposure and waterdeficit, especially when combined, whereas it was hastened by elevated CO2-temperature. Inthe last case, UV-B attenuated the effect of elevated CO2 and temperature. Changes in berryripening rates were associated with changes in photosynthetic performance.UV-B radiation and water deficit induced lower grape must acidity, mediated by increases inrelative skin mass or potassium levels rather than a decrease in organic acid concentration.In addition this increase in relative skin mass may have contributed to higher anthocyaninconcentration in the must. Grape berry skin flavonol and anthocyanin concentration wasincreased by UV-B, mainly due to the up-regulation of the structural (CHS, F3’H, FLS, UFGTand GST) and regulatory genes (MYBF1 and MYBA1) committed to their synthesis.Quantitative changes in flavonol concentration induced by UV-B were always associated withqualitative changes in flavonol profile (i.e. increased relative abundance of mono- anddisubstituted flavonols), as a result of the competition of FLS with flavonoid hydroxylases(F3’H and F3’5’H) for the same substrates. The independent up-regulation of FLS and F3’5’Hby UV-B radiation and water deficit, respectively, resulted in an intaractive effect on theflavonol B ring hydroxylation pattern. Under elevated CO2-temperature anthocyanin-sugaraccumulation was decoupled. However, UV-B partially alleviated this uncoupling by upregulatinganthocyanin biosynthesis and modulating berry ripening rates.UV-B radiation greatly influenced grapevine leaf physiology and berry composition. Theseresponses to UV-B were modulated, to a greater or lesser extent, by other factors linked toclimate change (water availability, atmospheric CO2 levels and temperature).

Plants are exposed to the stress of the environment throughout their entire live. They therefore had to develop sophisticated mechanisms to avoid or tolerate stress. In some plants, repeated exposure to stress increases tolerance, which is known as stress memory. The aim of this study is to assess stress memory in terms of oxidative stress level. Content of malondialdehyde (MDA), reactive oxygen species (ROS) and the electrolyte leakage were chosen as markers of oxidative stress. Fresh weight, plant height and water content were also evaluated. The level of oxidative stress was assessed in several genotypes of Solanum tuberosum subspecies, Andigena and Tuberosum. In vitro culture did not work well for the evaluation of stress memory, ex vitro partially did. The content of MDA in 66B was significantly lower than in cv. Lada (Tuberosum) in root (in vitro) and shoot (ex vitro) in control groups as well as during water shortage. Treatment of acclimation-inducing cold before the stress-inducing cold had a variable effect on growth and electrolyte leakage according to the culture conditions. In vitro, the effect on electrolyte leakage in roots of cv. Lada was positive, in leaves of 66B (Andigena) it was negative. In ex vitro, a positive effect was found on electrolyte leakage in 66B leaves, but at the...

Drosophila suzukii est une drosophile invasive en Europe, Amérique du Nord et Amérique du Sud. Contrairement aux autres espèces de drosophiles, les femelles parasitent les fruits mûrs que les larves consomment, engendrant d’importants dégâts sur les cultures fruitières. Les stratégies mises en place par cette espèce pour tolérer les températures hivernales sous nos latitudes sont encore peu comprises. Par conséquent, l’objectif de ma thèse était d’acquérir des connaissances fondamentales sur la thermotolérance de cette espèce, en m’intéressant notamment à la plasticité de la tolérance au froid et aux mécanismes physiologiques sous-jacents à l’acclimatation. J’ai évalué la thermotolérance basale de D. suzukii en soumettant des adultes et des pupes à un large panel de températures (froides et chaudes). Ces expérimentations ont permis de confirmer que cette espèce était intolérante au froid et que des températures supérieures à 32°C impactaient grandement sa survie. Par la suite, j’ai évalué la plasticité de sa tolérance au froid. Mes travaux ont permis de confirmer que sa thermotolérance était effectivement plastique, puisque l’utilisation de températures fluctuantes ou l’acclimatation permettaient de réduire sa mortalité lors d’expositions aux basses températures. L’acclimatation chez D. suzukii était corrélée à de nombreuses modifications physiologiques, telles que l’accumulation de cryoprotecteurs, un réajustement de la composition des phospholipides membranaires et des réserves lipidiques, une régulation des gènes liés à l’activité des transporteurs ioniques ainsi qu’un maintien de l’homéostasie métabolique. Ces modifications, également observées chez d’autres espèces d’insectes, pourraient être liées à l’augmentation de la tolérance au froid de D. suzukii, jouant probablement un rôle important dans sa survie hivernale et donc dans le succès de son invasion. Ces connaissances acquises sur sa thermobiologie contribueront sans doute à mieux cerner les limites physiologiques de cette espèce et prédire l’évolution de son invasion, ainsi que sa phénologie et les variations de populations au cours des saisons dans les zones déjà envahies. Mes résultats ouvrent également des perspectives intéressantes pour la mise en place de techniques de lutte intégrée contre D. suzukii. / Drosophila suzukii is an invasive pest in Europe, North and South America. Unlike other drosophilids, females oviposit in ripe fruits that larvae consume, provoking important damages on fruit productions. The overwintering strategies of this fly are yet poorly understood. Therefore, the aim of my thesis was to acquire new fundamental knowledge about the thermal biology of this fly, and more specifically the plasticity of its thermal tolerance and the physiological mechanisms underpinning cold acclimation. In order to define its basal thermal tolerance, adults and pupae were subjected to a large set of high and low temperatures. My data confirmed that this pest was chill susceptible, and showed that survival was greatly compromised during exposures above 32°C. Next, I evaluated its thermal tolerance plasticity. My data confirmed the high plasticity of its cold tolerance, as fluctuating thermal regimes and acclimation were able to decrease the mortality due to cold exposures. Acclimation in this species was correlate with several physiological adjustments, such as: cryoprotectant accumulation, remodeling of membrane phospholipids and lipidic reserves, upregulation of genes linked with activity of ionic transporters and maintenance of metabolic homeostasis. Those modifications (which are shared among temperate insect species) are likely linked with cold tolerance increase provoked by acclimation. Therefore, these physiological adjustments could play an important role in its overwintering success in Europe and Canada, which can facilitate its invasion in these regions. These new data will participate to a better understanding of its physiological limits, and are thus of importance for predicting the evolution of its invasion front and its phenology and demographic variations in invaded areas. My results are also of interest regarding the set-up of integrated pest management strategies against this fly.

The homeoviscous adaptation hypothesis states that the relative abundance of polyunsaturated fatty acids (PUFAs) in membrane phospholipids of ectothermic organisms decreases with increasing temperatures to maintainvital membrane properties. We reared Daphnia magna at 15°, 20°, and 25°C and increasing dietary concentrations of the long-chain PUFA eicosapentaenoic acid (EPA) to test the hypothesis that the well-documented increase in heat tolerance of high-temperature-reared Daphnia is due to a reduction in body PUFA concentrations. Heat tolerance was assessed by measuring the time to immobility at a lethally high temperature (Timm at 37°C), and whole body lipid fluorescence polarization (FP) was used as an estimate of membrane fluidity. At all rearing temperatures, EPA supplementation resulted in an increase in the relative abundance of EPA in body tissues, but only at 15° and 25°C did this result in a decrease in heat tolerance, and only at 20°C was this associated with an increase in membrane fluidity (i.e., decrease in FP). Overall, however, the degree of tissue fatty acid unsaturation correlated well with heat tolerance and FP. Our results support the homeoviscous adaptation hypothesis by showing that cold-reared Daphnia accumulate PUFAs within their body tissues and thus are more susceptible to heat than hot-reared Daphnia accumulating fewer PUFAs. However, our data also point out that further studies are required that elucidate the complex relationships between PUFA supply, membrane fluidity, and heat tolerance in ectotherms.

No description available.

Understanding how temperature affects biological systems is a central question in ecology and evolutionary biology. Anthropogenic climate change adds urgency to this topic, as the demise or success of species under climate change is expected to depend on how temperature affects important aspects of organismal performance, such as growth, development, survival and reproduction. Rates of biological processes generally increase with increasing temperature up to some maximal temperature. Variation in the slope of the initial, rising phase has attracted considerable interest and forms the focus of this thesis. I explore variation in growth rate-temperature relationships over several levels of biological organization, both between and within species, over individuals’ lifetime, depending on the ecological context and in relation to important life history characteristics such as generation length and winter dormancy.       Specifically, I examine how a clade of temperate damselflies have adapted to their thermal environment along a 3,600 km long latitudinal transect spanning from Southern Spain to Northern Sweden. For each of six species, I sampled populations from close to the northern and southern range margin, as well from the center of the latitudinal range. I reared larvae in the laboratory at several temperatures in order to measure indiviudal growth rates. Very few studies of thermal adaptation have employed such an extensive sampling approach, and my finding reveal variation in temperature responses at several levels of organization.       My main finding was that temperature responses became steeper with increasing latitude, both between species but also between latitudinal populations of the same species. Additional genetic studies revealed that this trend was maintained despite strong gene flow. I highlight the need to use more refined characterizations of latitudinal temperature clines in order to explain these findings. I also show that species differ in their ability to acclimate to novel conditions during ontogeny, and propose that this may reflect a cost-benefit trade-off driven by whether seasonal transitions occur rapidly or gradually during ontogeny.       I also carried out a microcosm experiment, where two of the six species were reared either separately or together, to determine the interacting effects of temperature and competition on larval growth rates and population size structure. The results revealed that the effects of competition can be strong enough to completely overcome the rate-depressing effects of low temperatures. I also found that competition had stronger effects on the amount of variation in growth rates than on the average value.       In summary, my thesis offers several novel insights into how temperature affects biological systems, from individuals to populations and across species’ ranges. I also show how it is possible to refine our hypotheses about thermal adaptation by considering the interacting effects of ecology, life history and environmental variation.

Le processus de dégradation anaérobie de la matière organique est un phénomène naturel largement répandu sur terre (ex. marais, lacs, rizières, systèmes digestifs d'animaux et humains). Une très grande diversité microbienne est entretenue durant ce processus, traduisant une diversité de voies métaboliques impliquées. Lorsqu'elle est complète, la digestion anaérobie aboutie à la formation de biogaz (mélange de méthane et de dioxyde de carbone). En termes de biotechnologie, le traitement par voie anaérobie de pollutions organiques permet de réduire le volume de déchets en générant du méthane valorisable sous plusieurs formes (électricité, chaleur, gaz naturel, biocarburant). Cependant, les digesteurs industriels sont optimisés pour un fonctionnement à 35°C ou à 55°C, ce qui nécessite un apport exogène d'énergie de maintenance. Ainsi, les travaux de thèse se sont intéressés à l'étude de la capacité d'adaptation de divers écosystèmes anaérobies industriels couvrant une variété de procédés et de conditions opératoires à convertir l'éthanol en biogaz à différentes températures. La première phase de l'étude avait pour but le conditionnement, en réacteurs de laboratoire d'écosystèmes à leur température d'origine avec un substrat facilement biodégradable (éthanol). Ensuite, les performances des communautés microbiennes (le potentiel méthanogène maximal et la cinétique de dégradation) ont été estimées sur un gradient de température de 5°C à 55°C en fioles. La phase de conditionnement des écosystèmes en réacteur batch a montré que la production de biogaz avoisinait la production théorique et que cette production s'accompagnait d'une diminution de la durée de réaction avec ajout successif du substrat. De plus, les cinétiques de production de biogaz obtenues les variaient fortement d'un écosystème à l'autre. Des profils d'empreintes moléculaires (CE-SSCP) des communautés bactériennes et archées ont été réalisés au début et à la fin du conditionnement. Ces profils de communauté ont été comparés entre eux par analyse en composante principale (ACP). Les populations bactériennes qui assuraient une performance efficiente étaient différentes de celles qui garantissaient une bonne capacité d'adaptation. Par ailleurs, le potentiel d'adaptation dépendait de la présence de populations d'Archaea méthanogènes bien spécifiques. En plaçant ensuite les écosystèmes conditionnés dans des conditions de température éloignées de la température d'origine, seuls les écosystèmes mésophiles se sont acclimatés aux températures psychrophiles. Comme attendu, l'activé spécifique maximale des méthanogènes était toujours obtenue à la température d'origine de l'écosystème. L'analyse des communautés bactériennes et archées à la fin de la période d'acclimatation a révélé que l'acclimatation des écosystèmes thermophiles et mésophiles à des températures plus faibles ne modifiait que légèrement la structure des communautés microbiennes. En revanche, des changements plus importants étaient obtenus lorsque la température d'incubation était augmentée par rapport à la température d'origine de l'écosystème. En résumé, l'étude de l'effet de la température d'incubation (de 5°C à 55°C) sur l'activité fermentaire et sur la structure des populations microbiennes est un bon modèle d'étude au laboratoire pour appréhender l'impact d'un facteur abiotique sur la dynamique structurelle et fonctionnelle d'une communauté microbienne complexe. / The process of anaerobic degradation of organic matter is a natural phenomenon widespread on Earth (eg, marshes, lakes, rice fields, digestive systems of animals and humans). A high microbial diversity is maintained during this process, reflecting a diversity of metabolic pathways involved. When complete, the anaerobic digestion accomplished in the formation of biogas (methane mixture and carbon dioxide). In terms of biotechnology, anaerobic treatment of organic pollution reduces the volume of waste and generates methane recoverable in several forms (electricity, heat, natural gas, biofuels). However, industrial digesters are optimized for operation at 35 ° C or 55 ° C, which requires exogenous energy maintenance. Thus, the thesis is interested in the study of the adaptability of various anaerobic ecosystems covering a variety of industrial processes and operating conditions to convert ethanol into biogas at different temperatures. The first phase of the study was to the conditioning, in laboratory reactors ecosystems to their original temperature with a readily biodegradable substrate (ethanol). Then, the performances of microbial communities (the maximum methanogenic potential and degradation kinetics) were estimated on a temperature gradient of 5 ° C to 55 ° C in glass bottles. The conditioning phase of the ecosystems in batch reactor showed that the biogas averaged theoretical production and this production was followed by a decrease in reaction time with successive addition of the substrate. In addition, the kinetics of the biogas obtained varied greatly from one ecosystem to another. Molecular fingerprinting profiles (CE-SSCP) of bacterial and archaeal communities were performed at the beginning and at the end of conditioning. These community profiles were compared with each other by principal component analysis (PCA). Bacterial populations that ensured efficient performance were different from those that ensured a good adaptability. In addition, the potential for adaptation depended on the presence of very specific methanogenic Archaea populations. When placing ecosystems conditioned in temperature away from the original temperature, only mesophilic ecosystems adapted to psychrophilic temperatures. As expected, specific methanogenic activity was always obtained at the original temperature of the ecosystem. Analysis of bacterial and archaeal communities at the end of the acclimation period revealed that acclimation thermophilic and mesophilic ecosystems to lower temperatures only modified slightly the structure of microbial communities. On the other hand, more significant changes were obtained when the incubation temperature was increased in comparison to the original temperature of the ecosystem. In summary, the study of the effect of incubation temperature (5 ° C to 55 ° C) on the fermentation activity and microbial population structure is a good model for laboratory study to understand the impact of abiotic factor on the structural and functional dynamics of a complex microbial community.

La recherche a été initiée par l'observation d'efflorescences de Planktothrix agardhii, une cyanobactérie dulçaquicole potentiellement toxique, dans deux étangs saumâtres, les étangs de l'Olivier et de Bolmon, avec dans ce dernier le déclin de l'espèce concomitamment à une augmentation de salinité. L'objectif de l'étude consistait à évaluer l'influence de la salinité du milieu sur la performance, l'hégémonie et la production de toxine de Planktothrix agardhii au sein de la communauté phytoplanctonique.La réalisation de suivis pluriannuels in situ couplés à des expérimentations en milieu contrôlé a permis de démontrer (i) la capacité d'acclimatation et d'adaptation à la salinité de Planktothrix agardhii, laquelle garantit sa suprématie et sa production toxinique en milieu saumâtre ; et (ii) la modification structurale et fonctionnelle de la communauté phytoplanctonique suite à une augmentation de salinité supérieure au seuil d'halotolérance de Planktothrix agardhii. La recherche témoigne ainsi de la versatilité des cyanobactéries qui renforce leur aptitude à être de bons compétiteurs, laissant présager leur persistance, la continuité de leurs nuisances, et leur expansion dans le futur. / The research was launched by the observation of Planktothrix agardhii blooms, a potentially toxic freshwater cyanobacterium, in two brackish ponds, the Olivier and Bolmon ponds, with in the latter, the concomitantly collapse of P. agardhii with an increase in salinity. The goal of the study was to assess the salinity influence on the performance, the dominance and the toxin production of P. agardhii within the phytoplankton community.The achievement of a long-term monitoring in situ combined with batch cultures experiments has demonstrated (i) the ability of P. agardhii to acclimate and adapt to salinity, which ensure its supremacy and its toxin production in brackish areas, and (ii) the structural and functional changes of the phytoplankton community with the exceeding of the salt-tolerance threshold of P. agardhii .The research reflects the cyanobacteria versatility that enhances their suitability for being good performers, suggesting their persistence along with their nuisances, and their expansion in the future.

De multiples événements sportifs majeurs vont se dérouler prochainement dans des environnements chauds voire tropicaux, justifiant l’intérêt scientifique actuel pour les questions associées à l’effet de la chaleur sur la performance en endurance. Il est admis que l’effort est subjectivement perçu comme plus difficile, en parallèle de la dégradation de la performance, à mesure que la température ambiante s’élève au cours d’une épreuve prolongée. Contrecarrer les effets délétères de la chaleur passe notamment par une exposition répétée dans des conditions écologiques ou simulées d’exercice (i.e. stratégie d’acclimatation). Toutefois, le décalage actuel entre les recommandations d’application issues d’études scientifiques et les conditions réelles de préparation et d’organisation des athlètes de haut niveau est à l’origine d’un faible engouement actuel pour de telles méthodes. Par conséquent, ce travail de thèse a eu pour ambition de répondre à certaines inconnues relatives à l’application de contenus d’entraînement classiques en ambiance chaude, en particulier (i) leur conséquence sur la capacité de performance de l’athlète et (ii) les moyens d’optimiser le contrôle et la régulation de la charge d’entraînement. Dans ce contexte, nous avons proposé à des sujets entraînés de soutenir la production de seuils de RPE (i.e. exercice à RPE fixe) dans un cadre expérimental puis d’entraînement. Au-delà de la validité de ce modèle, nous soutenons que la régulation volontaire de l’intensité dépend, au-delà de la perception de l’effort, de l’état émotionnel et de la motivation de l’individu. Lors d’une première étude, nous avons recherché à comparer les performances de solutions techniques pour le suivi de la température centrale en conditions écologiques (i.e. température gastro-intestinale vs. température frontale par capteur à annulation de flux). Les résultats obtenus ont validé l’usage du capteur à annulation de flux comme alternative pendant l’exercice, en dépit de l’absence de corrélation avec les mesures gastro-intestinales. Ceci suggère d’éventuelles perspectives en matière de contrôle de la température corporelle pendant l’exercice. La seconde étude s’est intéressée aux possibles conséquences de la répétition de sessions d’entraînement exigeantes et des contraintes logistiques d’un stage d’acclimatation (i.e. accumulation de fatigue mentale) sur la perception de l’effort et la performance. En dépit de l’absence d’effets combinés de la tâche cognitive préexercice et de la chaleur ambiante, les résultats tendent à démontrer le rôle-clé de la température cutanée et de la sensation de chaleur sous-jacente dans la régulation de la puissance soutenue à RPE-15 (chaud vs. tempéré : -0,022 vs. -0,008 W.kg-1.min-1). Enfin, la troisième étude suggère un potentiel intérêt de l’application de hautes intensités autorégulées, associée à une diminution du volume d’entraînement (-23%), lors d’une période d’acclimatation de courte durée (i.e. 5 jours). Le moindre effet observé, en comparaison d’un protocole à intensité fixe, sur la performance au cours d’un exercice de contre-la-montre (i.e. expérimental vs. fixe : 1,4 vs. 2,8 %) soulève toutefois l’importance du rapport volume-intensité dans la construction d’un protocole d’acclimatation. De manière générale, l’ensemble des résultats de cette thèse offrent des perspectives pour une individualisation et une adaptation spécifique à l’activité sportive des protocoles d’acclimatation à la chaleur. / The increasing number of major sport events that will take place in hot and/or tropical environments justify the current scientific interest in the effects of heat strain on endurance performance. During a prolonged self-paced exercise, it is well known that the subjectively perceived effort is higher as the ambient temperature increases and the performance level decreases. A repeated exposure to the heat in ecological and/or simulated exercise conditions may counteract the subsequent deleterious effects. However, the discrepancy between guidelines from scientific research and training priorities of well-trained athletes causes a lack of interest in these methods. Therefore, we aimed through the current thesis work to improve the current knowledge about heat acclimation strategies, more precisely about i) its effects on athlete’s performance capacity and ii) the optimization of training load monitoring and building. To shed light on these issues, we proposed to our trained and/or well-trained subjects to cycle at a fixed RPE first in an experimental framework, and then during a training program. We submit that the self-regulation of fixed-RPE exercise work rate depends not only of perceived exertion but also on emotional and motivational parameters. The first study aimed to compare the performance of technical devices for core temperature monitoring in ecological conditions (i.e. gastrointestinal point vs. forehead point from a zero-heat-flux sensor). Results showed that zero-heat-flux measurements might be considered as relevant during exercise. In this way, some opportunities may be considered for the monitoring of body temperature during field-based exercise. The second study investigated the subsequent effects of repeated strenuous training sessions and logistical constraints during a heat camp (i.e. higher mental fatigue) on perceived exertion and endurance performance. Despite the lack of combined effects from the prior cognitive task and the ambient condition during exercise, skin temperature and underlying heat sensation impact the linear decrease of power output at RPE-15 (hot vs. neutral: -0,022 vs. -0,008 W.kg-1.min-1). Lastly, the third study suggested an interest for self-regulated high intensities, associated with a decrease of the total exposure duration (-23%), during a short-term heat acclimation protocol (i.e. 5 days). However, the slighter effect on the improvement of performance in comparison with a classic constant-power training program, (HIT vs. constant-power: 1,4 vs. 2,8 %) provides scope for the building of training load (i.e. volume vs. intensity) in this context. In summary, the overall results of this thesis work open some perspectives for individualizing or adapting heat acclimation strategies to sport-specific conditions.

As propriedades bioquímicas da (Na+,K+)-ATPase branquial do siri eurialino Callinectes danae aclimatado à salinidade de 15 o/oo foram estudadas. A análise do gradiente de centrifugação em sacarose revelou a presença de um único pico entre 30-35% de sacarose, com uma boa correlação entre as atividades PNFFase a ATPase totais e (Na+,K+)-ATPase. A atividade residual observada na presença de ouabaína 3 mM sugere a presença de outros sistemas de enzimas atuantes. A eletroforese em condições desnaturantes nos microsomas de brânquias de C. danae em animais recém-coletados em salinidade de 33 o/oo (não aclimatados) e de aclimatados a salinidades de 15 e 33 o/oo por um período de 10 dias mostrou a presença de pequenas diferenças nos padrões eletroforéticos das diferentes amostras. A análise por Western blot mostrou um aumento significativo da proporção relativa da subunidade alfa da (Na+,K+)-ATPase em relação à proteína total na fração microsomal do tecido branquial de animais aclimatados à salinidade de 15 o/oo quando comparados aos animais aclimatados a 33 o/oo. Entretanto, proporções similares de subunidade alfa foram observadas para amostras de animais recém-coletados a salinidade de 33 o/oo e aclimatados a 15 o/oo. A estimulação da atividade (Na+,K+)-ATPase pelo ATP ocorreu através de uma curva de saturação monofásica apresentando interações sítio-sítio (nH=1,2), com V= 298,8 ± 16,7 U/mg, com K0,5 de 174,2 ± 9,8 uM. A estimulação da atividade ATPase da (Na+,K+)-ATPase por íons Mg2+ (V= 299,16 ± 14,06 U/mg; K0,5= 767,31 ± 36,06 uM), íons Na+ (V= 309,0 ± 15,8 U/mg; K0,5= 7,8 ± 0,4 mM), íons K+ (V= 300,6 ± 15,3 U/mg; K0,5= 1,63 ± 0,08 mM) e íons NH4+ (V= 345,1 ± 19,0 U/mg; K0,5= 6,0 ± 0,3 mM) ocorreu através de interações sítio-sítio. A atividade da enzima foi modulada sinergisticamente pelos íons K+ com atividade máxima variando de 300,6 ± 15,3 U/mg para 514,6 ± 26,2 U/mg, na ausência e na presença 50 mM de íons NH4+, respectivamente. Além disso, foi observado um significativo aumento na afinidade aparente da enzima pelo íon K+ da ordem de 10 vezes (diminuiu de 1,6 ± 0,08 mM para 0,157 ± 0,008 mM). Similarmente ao observado para os íons K+, o íon NH4+ estimulou sinergisticamente a atividade da enzima na presença de diferentes concentrações de íons K+. A estimulação da atividade da enzima pelo íon NH4+ também ocorreu através de interações cooperativas entre os sítios. Embora tenha sido observado um aumento da atividade específica da enzima de 345,1 ± 19,0 U/mg para 516,8 ± 27,9 U/mg, não foram observadas variações significativas nos valores de nH e K0,5 com o aumento da concentração de íons K+. A ouabaína inibiu cerca de 90% da atividade ATPase total. A inibição pela ouabaína apresentou valor de KI de 45,09 ± 2,51 uM. O ortovanadato também inibiu atividade (Na+,K+)-ATPase na mesma faixa (90%) através de uma curva de inibição monofásica, com valor de KI da ordem de 1,31 ± 0,06 uM. O emprego de bafilomicina A1, tapsigargina e teofilina, juntamente com a ouabaína, na atividade ATPase total descartam a presença de V-ATPase, Ca2+-ATPase ou fosfatase, respectivamente. Apesar da inibição por oligomicina corresponder a menos de 3,7%, esse valor aparentemente sugere a presença de uma F0F1-ATPase. Além disso, a inibição por ácido etacrínico, em conjunto com os experimentos de estimulação por da atividade ATPase da enzima por íons Na+ sugere fortemente a presença de uma K+-ATPase. A (Na+,K+)-ATPase hidrolisou o substrato PNFF obedecendo à cinética Michaeliana com velocidade de V= 102,9 ± 4,3 U/mg e KM= 1,7 ± 0,1 mM. Já a estimulação da atividade K+-fosfatase da enzima por íons Mg2+ (V= 93,7 ± 2,3 U/mg; K0,5= 1,40 ± 0,03 mM), K+ (V= 94,9 ± 3,5 U/mg; K0,5= 2,9 ± 0,1 mM) e NH4+ (V= 106,2 ± 2,2 U/mg; K0,5= 9,8 ± 0,2 mM) seguiu uma cinética cooperativa, sugerindo a presença de múltiplos sítios de ligação. Entretanto, a atividade K+-fosfatase não foi estimulada sinergísticamente na presença de íons K+ mais NH4+. Os íons sódio (KI= 22,7 ± 1,7 mM) e ortovanadato (KI= 28,1 ± 1,4 nM) inibiram completamente a atividade fosfatase total através de uma única curva de inibição. / The biochemical properties of the (Na+,K+)-ATPase from the gill microsomal tissue of the euryhaline, marine, swimming crab Callinectes danae, acclimated to 15 0/00 salinity, were investigated. Sucrose gradient centrifugation analyses revealed a unique peak, between 30-35% sucrose, coincident with the total PNPPase, ATPase, and (Na+,K+)-ATPase activities. The residual activity observed in the presence of 3 mM ouabain suggests the existence of other enzyme systems. Electrophoresis under denaturing conditions, using material from fresh-caught crabs (33 o/oo salinity, not acclimated), and from crabs acclimated to 15 or 33 o/oo salinity, for 10 days, revealed differences in migration pattern. Western blot analyses showed a significant increase in the amount of (Na+,K+)-ATPase alpha-subunit relative to total protein, for crabs acclimated to 15 o/oo compared to those acclimated to 33 o/oo salinity. However, the proportion of alpha-subunit in samples from fresh-caught crabs acclimated to 33 o/oo and those acclimated to 15 o/oo salinity was similar. (Na+,K+)-ATPase activity was stimulated by ATP and showed a single saturation curve, exhibiting site-site interactions (nH=1.2), with V= 298.8 ± 16.7 U/mg, and K0.5= 174.2 ± 9.8 uM. Stimulation of the ATPase activity by Mg2+ (V= 299.16 ± 14.06 U/mg; K0.5= 767.31 ± 36.06 uM), Na+ (V= 309.0 ± 15.8 U/mg; K0.5= 7.8 ± 0.4 mM), K+ (V= 300.6 ± 15.3 U/mg; K0.5= 1.63 ± 0.08 mM) and NH4+ ions (V= 345.1 ± 19.0 U/mg; K0.5= 6.0 ± 0.3 mM) occurred through site-site interactions. (Na+,K+)-ATPase activity was synergistically modulated by K+ ions, maximum activity varying from 300.6 ± 15.3 U/mg to 514.6 ± 26.2 U/mg, in the absence and presence of 50 mM NH4+ ions, respectively. K+ ions induced a 10-fold increase in enzyme apparent affinity (from 1.6 ± 0.08 mM to 0.157 ± 0.008 mM). As for K+ ions, NH4+ synergistically stimulated enzyme activity in the presence of variable K+ concentrations. The stimulation by NH4+ ions exhibited cooperative, site-site interactions. Although an increase in specific activity from 345.1 ± 19.0 U/mg to 516.8 ± 27.9 U/mg was seen, no significant changes in nH and K0.5 were observed. Ouabain inhibited total ATPase activity by about 90%, showing a KI= 45.09 ± 2.51 uM. Orthovanadate also inhibited the (Na+,K+)-ATPase with a KI of 1.31 ± 0.06 uM. Although the inhibitory effect of oligomycin was minimal (3.7%), this inhibition may suggest F0F1-ATPase activity. The inhibition by ethacrynic acid, in association with Na+ ion stimulation of the ATPase activity, suggests the presence of a K+-ATPase. The (Na+,K+)-ATPase hydrolyzed PNPP (K+-phosphatase activity) obeying Michaelian kinetics, with V= 102.9 ± 4.3 U/mg and KM= 1.7 ± 0.1 mM. The stimulation of K+-phosphatase activity by Mg2+ (V= 93.7 ± 2.3 U/mg; K0.5= 1.4 ± 0.03 mM), K+ (V= 94.9 ± 3.5 U/mg; K0.5= 2.9 ± 0.1 mM), and NH4+ ions (V= 106.2 ± 2.2 U/mg; K0.5= 9.8 ± 0.2 mM) following cooperative kinetics, suggests multiple binding sites. K+-phosphatase activity, however, was not synergistically stimulated by K+ and NH4+. Sodium ions (KI= 22.7 ± 1.7 mM), and orthovanadate (KI= 28.1 ± 1.4 nM) totally inhibited the total phosphatase activity.