Made available in DSpace on 2015-07-22T20:49:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-03-31 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Introdução: Os pacientes com Miopatia Mitocondrial (MM) e Oftalmoplegia Externa Progressiva (OEP) apresentam disfunção na cadeia respiratória com incapacidade de aumentar a extração de oxigênio muscular e sintetizar ATP aerobicamente, levando à intolerância ao esforço e lentificação da cinética do O2. Quando a extração de oxigênio é comprometida, na tentativa de se manter o consumo de oxigênio muscular, uma hipótese é que estes pacientes poderiam aumentar compensatoriamente a oferta de oxigênio, apresentando, assim, uma resposta hipercinética cardiovascular e ventilatória. Por outro lado, alguns indícios de menor oferta de oxigênio foram encontrados em pacientes com MM, como um menor fluxo sanguíneo muscular no antebraço e uma maior capacidade de produção de ATP após a suplementação de oxigênio. A cinética do O2 na fase de recuperação (REC) nos fornece, assim, subsídios quanto ao pagamento do débito de oxigênio tecidual e ao reabastecimento do estoque de oxigênio sanguíneo após o exercício. Ao nosso conhecimento, a cinética do O2 REC nunca foi avaliada nos pacientes com MM, assim como a integração desta variável com as respostas não-invasivas cardiovasculares e de extração de oxigênio muscular. Objetivo: contrastar as dinâmicas da oferta e da utilização de oxigênio na recuperação do exercício em pacientes com MM; e identificar os principais mecanismos fisiopatológicos da intolerância ao esforço nestes indivíduos. Métodos: Foram avaliadas em 12 pacientes com MM e 12 controles saudáveis, as cinéticas de recuperação: (i) do O2 pulmonar, (ii) da variação na concentração da deoxiemoglobina ([HHb], mensurada pela espectroscopia de raios quasi infravermelhos - NIRS) no vasto lateral, (iii) do débito cardíaco (DC) por bioimpedância transtorácica, após um teste de carga constante de alta intensidade (70% da carga máxima atingida em teste incremental prévio) até o limite da tolerância em cicloergômetro. Resultados: Foram observadas cinéticas mais lentas de reoxigenação da [HHb], ([HHb] = 43,7 ± 21,2 vs 27,5 ± 6,7) e do O2 ( O2 = 58,1 ± 25,1 vs 38,8 ± 7,6) nos pacientes com MM em relação aos controles, respectivamente. Estas respostas foram associadas a uma cinética de DC mais rápida em relação ao O2, nos pacientes comparados aos controles (T½DC * 1,44 / O2 = 1,3 ± 0,4 vs 1,7 ± 0,6). Conclusão: Os pacientes com MM na forma OEP apresentam, na recuperação do exercício de alta intensidade, um pagamento elevado do débito de oxigênio contraído no exercício e reoxigenação mais lenta da [HHb]. Estas respostas, associadas à cinética mais rápida do DC em relação ao O2 são indícios de que possa haver um déficit no transporte de oxigênio microvascular, além do comprometimento mitocondrial característico desta doença. / Background: Mitochondrial Myopathy patients (MM) and Progressive External Ophthalmoplegia (PEO) present with respiratory chain dysfunction and inability to increase muscle oxygen extraction and aerobic ATP synthesis, leading to exercise intolerance and slower O2 kinetics. When oxygen extraction is impaired, in an attempt to maintain muscle oxygen uptake, these patients could increase oxygen delivery, thus exhibiting a hyperkinetic cardiovascular and ventilatory response. On the other hand, some evidence of oxygen delivery impairment was found in MM patients, such as a decrease in muscle blood flow in the forearm and a greater capacity for ATP production after oxygen supplementation. Recovery O2 kinetics provides information on tissue oxygen debt repayment and oxygen blood store replenishment after exercise. To our knowledge, recovery O2 kinetics has never been evaluated in MM patients, as well as its integration with the non-invasive cardiovascular and muscle reoxygenation responses. Objective: to contrast oxygen delivery and utilization dynamics on exercise recovery of MM patients and to identify the main pathophysiologic mechanisms of exercise intolerance in these subjects. Methods: Were evaluated in 12 MM patients and 12 healthy controls, the recovery kinetics of: (i) O2 (ii) deoxyhemoglobin variation ([HHb], measured by near-infrared spectroscopy - NIRS) in vastus lateralis, (iii) cardiac output (CO) by transthoracic bioimpedance, after a high-intensity constant work rate test (70% of maximal workload in a previous incremental test) to the limit of tolerance in a cycle ergometer. Results: We detected slower kinetics for [HHb] ([HHb] = 43.7 ± 21.2 vs. 27.5 ± 6.7) and for O2 ( O2 = 58.1 ± 25.1 vs. 38.8 ± 7.6) in MM patients compared to controls, respectively. Additionally, these responses were associated with a faster recovery CO kinetics in relation to O2 kinetics in MM patients compared to controls (T½DC*1,44 / O2 = 1,3 ± 0,4 vs. 1,7 ± 0,6). Conclusion: Patients with MM and PEO present with a higher oxygen debt and slower reoxygenation kinetics in the recovery of a high-intensity exercise test. Those responses were associated with a faster CO recovery in relation to O2 kinetics, indicating a microvascular oxygen transport deficit, besides the characteristic mitochondrial impairment observed in these patients. / TEDE / BV UNIFESP: Teses e dissertações

O trabalho apresenta uma investigação sobre a influência da umidade atmosférica no processo de absorção de oxigênio atmosférico por corpo de água através de sua superfície líquida livre. Os experimentos realizados em laboratório consistem na reoxigenação de uma massa de água, contida em um tanque cilíndrico, submetida à agitação mecânica. A atmosfera que reabastece de oxigênio a massa líquida é controlada e os ensaios são conduzidos com vários níveis de umidade do ar, o que provoca diferentes taxas de evaporação. Simultaneamente, determinam-se os coeficientes de reoxigenação, K2 (h-1), em cada experimento. Investiga-se neste trabalho a correlação entre esses dois parâmetros. O processo de reoxigenação é analisado inicialmente com o uso de metodologia clássica baseada em modelos matemáticos tradicionais (possivelmente inadequados). Propõe-se neste trabalho, um modelo original para essa mesma análise. / The influence of the atmospheric humidity on the process of absorption of oxygen by a water body through its free surface is presented herein. Laboratory experiments were run for the reoxygenation of a water mass kept in a cylindrical vessel under mechanical agitation. The quality of the atmospheric air transferring oxygen to water is controlled and the tests are run with several levels of air humidity, what renders different rates of evaporation. The corresponding coefficients of reaeration K2 (h-1) are determined for each experiment. The correlation between these two latter parameters is investigated. The process of reoxygenation is analyzed initially using a classical methodology based on traditional mathematical models (possibly inadequate). An original model for the same analysis is proposed in this work.

O parâmetro que determina o processo de transferência de oxigênio através da interface ar-água em escoamentos com superficie livre é o coeficiente de reaeração superficial, \'K IND.2\'. Existe um grande número de equações na literatura científica que realizam a previsão do coeficiente de reaeração, no entanto, a maioria dessas equações considera apenas algumas características hidráulicas do corpo de água como a velocidade e a profundidade, e ainda, como em sua maioria são de natureza empírica, acabam sendo adequadas para corpos de água específicos. Além destes dois parâmetros (velocidade e profundidade do escoamento), existem outros doze que são considerados importantes para estudos de reoxigenação em águas superficiais. É neste contexto que o objetivo deste trabalho foi estudar em condições de laboratório a influência da rugosidade do leito de um canal sobre o coeficiente de reaeração superficial. Além da rugosidade, foram variadas as condições de velocidade média do escoamento e profundidade da lâmina de água. A variação da rugosidade foi realizada através da implementação de obstáculos com geometria conhecida, dispostos transversalmente ao escoamento, e denominados como macro-rugosidades. Os resultados demonstraram evidências de que o coeficiente de reaeração superficial é controlado de forma significativa pela rugosidade do leito do canal. Foram desenvolvidos dois modelos matemáticos de natureza semi-empírica para a previsão do \'K IND.2\', os quais incorporam variáveis hidrodinâmicas e geométricas consideradas relevantes para o processo de transferência de massa na interface ar-água. / The parameter that determines the process of oxygen transfer through the air-water interface in flows with free surface is the superficial reaeration coefficient, \'K IND.2\'. There are a large quantity of equations in the cientific literature that perform the prevision of the reaeration coefficient; however, most of them consider only some of the hydraulic characteristics of water body; for example, the velocity and the depth. Most of these equations have empirical origin and are adequate just for a specific water body. In addition to these two parameters (velocity and depth of flow), there are another twelve important parameters in reoxygenation studies in superficial waters. In this context, the aim of this work was to study the influence of bed roughness of a channel on the superficial reaeration coefficient, under laboratory conditions. Besides the roughness, the conditions of mean velocity of the flow and depth of water were varied. The variation of roughness was done with implementation of obstacles with known geometry, arranged transversely to the flow direction, and called macro-roughness. The results have shown evidences that the surface reaeration coefficient is controlled significantly by the roughness of the channel bed. Two mathematical models of semi-empirical origin for the \'K IND.2\' prevision were developed, which incorporate hydrodynamic and geometric variables considered relevant in the process of mass transfer in air-water interface.

A reoxigenação de água residuária é tratada na literatura técnica de maneira análoga à reoxigenação na água limpa; os efeitos do consumo de oxigênio pela população bacteriana ativa são quantificados pela introdução dos parâmetros alfa e beta. Alfa corrige o coeficiente volumétrico de transferência de massa do oxigênio e beta corrige a concentração de saturação do oxigênio dissolvido na água. Com o presente estudo pretende-se tornar explícitos os parâmetros cinéticos envolvidos no processo, de modo a fornecer base conceitual para um melhor entendimento do fenômeno. A pesquisa é feita a partir de trabalho experimental, seguido de modelagem matemática conceitual. No entanto, no lugar dos microrganismos como sumidouro de oxigênio dissolvido foi utilizado o borbulhamento sub-superficial de gás nitrogênio, em processo de stripping. Um processo físico, portanto, substitui o processo bioquímico no papel de retirar o oxigênio dissolvido no seio líquido. O modelo matemático proposto ajustou-se bem aos dados experimentais, sendo que a sua validação pôde ser obtida através das análises estatísticas realizadas. / The reoxygenation in wastewater is seen in the technical literature in an analogous way as the reoxygenation of clean water. The effects of the oxygen consumed by active bacteria population are quantified with the introduction of the parameters alpha and beta. Alpha corrects for the volumetric mass transfer coefficient and beta corrects for the dissolved oxygen saturation concentration. The present research intends to make explicit the kinetic of the process, in order to supply conceptual support to a better understanding of the phenomenon. The research is done through experimental work, followed by conceptual, mathematical modeling. Stripping with nitrogen is used as an oxygen sink instead of microorganisms. Thus, a physical process replaces the biochemical process. The proposed mathematical model fitted well to the experimental data, and its validation could be obtained through the statistical analyses done.

En préservation-transplantation rénale, l’ischémie-reperfusion (IR) induit inflammation, fibrose, dysfonction et perte du greffon. Les événements d’IR sont de mieux en mieux identifiés, mais leurs complexité limite prédiction et thérapeutique. A partir d’une analyse bibliographique détaillée, nous proposons un modèle par agents de la réponse rénale aux lésions d’IR au niveau cellulaire/tissulaire, réalisé avec l'outil de modélisation NetLogo.Dans un premier temps, nous développons un modèle dynamique de l’oxygénation du cortex rénal, avec apport et diffusion de l’oxygène (O2), et consommation dépendant de la filtration. Nous adaptons ce modèle à l’état stationnaire pour l’O2, puis nous couplons les niveaux de PO2 à l'état énergétique (ATP) des cellules épithéliales et endothéliales (avec voies aérobie et anaérobie). Le statut de viabilité cellulaire est lié au niveau d'ATP, aboutissant à une représentation semi-phénoménologique de la réparation/survie vs apoptose/nécrose. Enfin, nous explorons le destin cellulaire et tissulaire lors d’IR simulées, avec l’ajout progressif d’éléments clefs de l’inflammation (invasion tissulaire par leucocytes, signaux lésionnels, phagocytose) et de la fibrose (fibroblastes, collagène). L’évolution du modèle vers la résolution de l’inflammation/régénération du tissu ou vers la fibrose tissulaire est observée selon les conditions imposées (durée/intensité, ischémie vs hypoxémie).Cette construction constitue le premier modèle des effets de l’oxygénation sur la dynamique cellulaire-tissulaire de l’inflammation-fibrose rénale en réponse à l’IR. A terme, elle permettra d'aborder clinique et thérapeutique de la conservation-transplantation rénale. / In renal preservation-transplantation, ischemia-reperfusion (IR) causes graft inflammation and fibrosis, dysfunction and loss. Events involved in IR injury grow identified, but their intricacy hampers prediction and therapeutics. Based on a detailed bibliographical analysis, we propose an Agent-Based Model of renal response to IR injury at cell and tissue levels, created with the modeling tool NetLogo.First, we develop and validate a dynamic model of the oxygenation of the renal cortex, featuring blood perfusion, oxygen diffusion, and oxygen consumption (driven by sodium filtered load and transport). We then adapt this model to oxygen steady-state, and PO2 level is coupled to energetic status (ATP) in epithelial and endothelial cells (aerobic and anaerobic pathways). Cell viability is coupled to ATP level, leading to a semi-phenomenological representation of repair/survival versus apoptosis/necrosis. Finally, we explore (and verify) cell and tissue fate during simulated IR sequences, with the gradual addition of key elements of inflammation (leukocytes infiltration, injury signals, phagocytosis) and fibrosis (fibroblasts, collagen). Model evolution toward the resolution of inflammation/tissue regeneration or toward tissue fibrosis is observed along imposed conditions (duration/intensity, ischemia vs hypoxemia). Results are compared to experiments from our laboratory.This construction is the first model of the effects of oxygenation on cell-tissue dynamics during renal inflammation-fibrosis response to IR. Ultimately, it will allow to address clinical and therapeutic aspects of renal transplantation and conservation.

La maladie coronaire d’origine ischémique reste l’une des principales causes de mortalité dans le monde industrialisé.Le traitement de l’ischémie aiguë du myocarde est entré dans une nouvelle ère où la mortalité peut être diminuée de moitié en utilisant des procédures qui permettent un retour rapide du débit sanguin dans la zone ischémique du myocarde: la revascularisation.Cette reperfusion entraîne des complications appelées lésions de la reperfusion qui ont été décrites pour la 1ère fois par Jennings et al., en 1960. Le développement de stratégies cardioprotectrices associées à la reperfusion constitue un besoin majeur en clinique afin d’améliorer la fonction myocardique, de diminuer l'incidence des arythmies, de retarder l'apparition de la mort cardiomyocytaire et de limiter la taille de l’infarctus du myocarde lors de l'ischémie/reperfusion (I/R).La découverte de 2 formes principales de mécanismes cardioprotecteurs endogènes, a encouragé la recherche de nouveaux moyens pharmacologiques capables de protéger le myocarde ischémié/reperfusé et a développé nos connaissances sur les bases moléculaires des lésions et de la survie cellulaire au cours des processus d’I/R.L’étude des mécanismes responsables de l’induction de la mort cellulaire a permis de mettre en évidence le rôle joué par la mitochondrie et l’augmentation de la perméabilité de ses membranes, induite par la formation/ouverture d’un pore au niveau des points de contacts entre les membranes mitochondriales;ce pore a été appelé « pore de transition de la perméabilité mitochondriale » (mPTP).L’inhibition de l’ouverture de ce pore apparaît comme une stratégie privilégiée pour protéger le myocarde.Des études ont montré que les espèces réactives d’oxygène (EROs) jouent un rôle majeur dans les lésions de l’I/R et dans l’ouverture du mPTP.Il existe peu d’informations claires sur le seuil et la période de production (ischémie et/ou reperfusion) des EROs qui conduisent à l’ouverture du mPTP.Nous avons mis au point un modèle cellulaire d’hypoxie/réoxygénation (H/R) afin d’établir une relation causale entre la production d’EROs, l’ouverture du mPTP et la mort cellulaire tout en explorant le rôle de différents types d’EROs.Ce modèle d’H/R nous a permis de mesurer en temps réel et simultanément la production des EROs, l’ouverture du mPTP et la mort cellulaire.Nous avons montré que la production des EROs débute pendant la période d’hypoxie et qu’elle est directement liée à l’augmentation du temps d’hypoxie.Cette production d’EROs à l’hypoxie, plus particulièrement de radicaux hydroxyles et de peroxyde d’hydrogène, a été directement relié, à l’ouverture du mPTP et à la mort cellulaire lors de l’H/R.Nous avons utilisé ce modèle pour étudier le mécanisme d’action de deux stratégies pharmacologiques cardioprotectrices, un nouveau ligand de la protéine translocatrice mitochondriale (TSPO), le TRO 40303, et l’activation de la voie RISK par la morphine. Nous avons ainsi montré que (1) les propriétés cardioprotectrices du TRO40303 sont associées à une inhibition de l’ouverture du mPTP, ce qui n’avait pas pu être démontré au moyen d’expériences réalisées ex vivo et (2) l’activation de la voie RISK par la morphine, qui aboutit à une limitation de la taille d’infarctus associée à une amélioration des fonctions respiratoires mitochondriales, entraîne également une inhibition de l’ouverture du mPTP et un retard de la mort cellulaire des cardiomyocytes isolés soumis à une H/R.La suite de ce travail sera de rechercher si l’inhibition du stress oxydant peut constituer un mécanisme commun aux deux stratégies pharmacologiques cardioprotectrices en utilisant notre modèle d’H/R.Il serait possible d’étendre notre modèle à des animaux génétiquement modifiés pour appréhender les phénomènes impliqués dans cette activité antioxydante.A plus long terme, il sera nécessaire d’approfondir nos connaissances sur la production d’EROs pendant l’I/R en recherchant plus spécifiquement l’origine de cette production. / Ischemic coronary artery disease remains one of the main causes of mortality in the industrialized countries. The treatment of acute myocardial ischemia entered a new era where mortality can be reduced by 50% using revascularization procedures that allow a rapid return of blood flow to the ischemic area. However, this reperfusion leads to complications known as lethal reperfusion induced injury that have been described for the first time by Jennings et al., in 1960. It became crucial to develop cardioprotective strategies in combination with early reperfusion in order to improve myocardial function, to reduce the incidence of arrhythmias, to delay the onset of cardiomyocytes death and to limit the extension of infarct size following reperfusion. The discovery of two major forms of endogenous cardioprotective mechanisms, which consist of the realization of short cycles of ischemia/reperfusion (I/R) prior to a long period of ischemia (ischemic preconditioning) or before reperfusion after the long period of ischemia (Ischemic Postconditioning), encouraged the search for new pharmacological tools to protect the ischemic myocardium to develop our knowledge on the molecular mechanisms of lethal reperfusion injury and cell survival in the I/R process.The study of cell death mechanisms has highlighted the crucial role of the mitochondria and more specifically the increase in mitochondrial membrane permeability following I/R.One reason for increasing permeability is the formation/opening of a pore at mitochondrial membranes contact sites at reperfusion.This pore has been called "the mitochondrial permeability transition pore" (mPTP). Inhibition of this pore opening has been presented as a main strategy to protect the myocardium.Many studies have shown that reactive oxygen species (ROS) play a major role in I/R injury and mPTP opening, but there is very few information to date about the threshold and the period of ROS production (ischemia and/or reperfusion) that lead to mPTP opening.We designed a cellular model of hypoxia/reoxygenation (H/R) to establish a causal relationship between ROS production, mPTP opening and cell death while exploring the role of different types of ROS.This H/R model used freshly isolated adult rat cardiomyocytes and allowed us to measure online and simultaneously ROS production, mPTP opening and cell death. We have demonstrated that ROS production starts during the period of hypoxia and thisproduction is directly linked to the increase in the duration of hypoxia.This ROS production during hypoxia has been, for the first time, directly related to mPTP opening and cell death following H/R.We used this model to study the mechanism of action of two cardioprotective strategies, a new ligand of the mitochondrial translocator protein (TSPO), TRO 40303 and a RISK (Reperfusion Injury Salvage Kinase) pathway activator, morphine. We have shown that (1) the cardioprotective properties of TRO40303 were associated with inhibition of mPTP opening, a mechanism that could not be demonstrated using ex vivo experiments and (2) morphine that provoked infarct size limitation associated with an improvement of mitochondrial respiratory functions through RISK pathway activation, also inhibited mPTP opening and delayed cell death of isolated cardiomyocytes subjected to H/R.Finally, a question comes into sight whether the inhibition of oxidative stress may be a common mechanism to both cardioprotective pharmacological strategies that we have described using our H/R model. To do this, it would be possible to extend our model to genetically modified animals specifically adapted to understand the phenomena involved in antioxidant activity.On long-term, it will be necessary to develop our knowledge on ROS production during I/R by looking for the origin of this production, more precisely the role of the mitochondria and the effect of other reactive species in order to target the treatment and to develop new cardioprotective strategies.

No description available.

In der Pathogenese der Temporallappenepilepsie wird kindlichen hippokampalen Schädigungen eine wesentliche Rolle zugeschrieben. Epileptische Krämpfe und perinatale Asphyxie sind zwei häufige Ursachen dieser Schädigungen. Anhaltende epileptiforme Aktivität im Niedrig-Mg2+-Modell als einer experimentellen Form epileptischer Krämpfe führt in organotypischen hippokampalen Schnittkulturen (OHSK) der Ratte, die als Ersatzsystem des kindlichen Hippokampus verwendet werden, zu Zellschäden. Während dieser Untersuchungen ergab sich der Verdacht auf eine zusätzlich schädigende Wirkung erhöhter Sauerstoffspannungen. In meiner ersten Versuchsreihe konnte ich nachweisen, dass erhöhte Sauerstoffspannungen (60 %, 95 %) verglichen mit 20%-Sauerstoffspannung zu reversiblen und irreversiblen Zellschäden in OHSK führen. Die Zellschäden wurden über Veränderungen reizinduzierter Feldpotentiale, d.h. Abnahme der Amplitude, Zunahme der Latenz und Zunahme des Doppelpulsindex, sowie über die Propidium Jodid (PJ)-Fluoreszenzintensität bestimmt. In der zweiten Versuchsreihe konnte gezeigt werden, dass erhöhte Sauerstoffspannungen auch nach einer Hypoxie im Sinne einer hyperoxischen Reoxygenierung verglichen mit normoxischer Reoxygenierung vermehrt Zellschäden in OHSK zur Folge haben. In der dritten Versuchsreihe konnte ich ausschließen, dass erhöhte Sauerstoffspannungen eine notwendige Bedingung für Zellschäden infolge anhaltender epileptiformer Aktivität sind. Um die zellschädigende Rolle von Spreading Depressions (SDs), die während epileptiformer Aktivität auftreten, zu bestimmen, wurde in der vierten Versuchsreihe eine Methode etabliert, SD-ähnliche Ereignisse isoliert und zuverlässig in normoxischen OHSK auszulösen. Auf diese Weise wiederholt ausgelöste SD-ähnliche Ereignisse führten zu Zellschäden, bestimmt über die Veränderung elektrophysiologischer Eigenschaften von SD-ähnlichen Ereignissen, Abnahme der Feldpotentialamplitude und PJ-Fluoreszenzintensität. / Hippocampal damage during infancy is thought to play an important role in the pathogenesis of temporal lobe epilepsy. Epileptic seizures and perinatal asphyxia are two frequent causes of these damages. Sustained epileptiform activity induced in the low Mg2+-model of epileptic seizures leads to cell damage in organotypic hippocampal slice cultures (OHSC) of the rat, which are used as a surrogate for the infantile hippocampus. During a previous study utilising this model the suspicion arose that increased oxygen tension could have an additional damaging effect. My first series of experiments proved that increased oxygen tension (60 %, 95 %) lead to reversible and irreversible cell damage in OHSC compared to 20%-oxygen tension. Cell damage was determined by alterations of evoked field potentials, i.e. decrement of amplitude, increment of latency and paired pulse index, as well as by propidium iodide fluorescence. The second series of experiments showed that increased oxygen tension applied after an hypoxic period (hyperoxic reoxygenation) result in augmented cell damage compared to normoxic reoxygenation. With the third series of experiments it could be excluded that increased oxygen tension is an essential condition for the occurrence of cell damage due to sustained epileptiform activity. In order to elucidate the damaging role of spreading depressions (SD), which emerge during epileptiform activity, a method was established in the fourth series of experiments that allowed the reliable induction of SD-like events in normoxic OHSC. Repetitive SD-like events induced by this method led to cell damage, assessed by alterations of electrophysiological characteristics of SD-like events, decrement of evoked field potential amplitude and propidium iodide fluorescence.

No description available.

Fish encounter daily cycles of hypoxia in the wild, but the physiological strategies for coping with repeated cycles of normoxia and hypoxia (intermittent hypoxia) are poorly understood. Contrastingly, the physiological strategies for coping with continuous (constant) exposure to hypoxia have been studied extensively in fish. The main objective of this thesis was to understand how Fundulus killifish cope with a diurnal cycle of intermittent hypoxia, an ecologically relevant pattern of aquatic hypoxia in the natural environment. To do this, I characterized the effects of intermittent hypoxia on hypoxia tolerance, oxygen transport, metabolism, and the oxidative stress defense system of killifish, and compared these effects to fish exposed to normoxia, a single cycle of hypoxia-normoxia, and constant hypoxia. Specifically, I studied the following topics: (i) how acclimation to intermittent hypoxia modifies hypoxia tolerance, and the hypoxia acclimation response of Fundulus heteroclitus (Chapter 2), (ii) metabolic adjustments occurring during a hypoxia-reoxygenation cycle (Chapter 3), (iii) how acclimation to intermittent hypoxia alters O2 transport capacity and maximal aerobic metabolic rate (Chapter 4), (iv) the effects of hypoxia and reoxygenation on reactive oxygen species and oxidative stress (Chapter 5), and (v) variation in hypoxia tolerance and in the hypoxia acclimation responses across Fundulus fishes (Chapter 6). Killifish rely on a unique and effective physiological strategy to cope with intermittent hypoxia, and that this strategy is distinct from both the response to a single bout of acute hypoxia-reoxygenation (12 h hypoxia followed by 6 h reoxygenation) and to chronic exposure to constant hypoxia (24 h hypoxia per day for 28 d). Key features of the acclimation response to intermittent hypoxia include (i) maintenance of resting O2 consumption rate in hypoxia followed by a substantial increase in O2 consumption rate during recovery in normoxia, (ii) reversible increases in blood O2 carrying capacity during hypoxia bouts, (iii) minimal recruitment of anaerobic metabolism during hypoxia bouts, and (iv) protection of tissues from oxidative damage despite alterations in the homeostasis of reactive oxygen species and cellular redox status. Of these features, (i) is unique to intermittent hypoxia, (ii) also occurs in fish exposed to acute hypoxia-reoxygenation, and (iii) and (iv) are observed in both fish acclimated to intermittent hypoxia as well as those acclimated to constant hypoxia. This is the most extensive investigation to date on how fish cope with the energetic and oxidative stress challenges of intermittent hypoxia, and how these responses differ from constant hypoxia. This thesis adds substantial insight into the general mechanisms by which animals can respond to an ecologically important but poorly understood feature of the aquatic environment. / Dissertation / Doctor of Philosophy (PhD) / Oxygen levels in the aquatic environment are dynamic. Many fishes routinely encounter changes in oxygen content in their environment. However, we have very little understanding of how cycles between periods of low oxygen (hypoxia) and periods of high oxygen (normoxia) affect the physiology of fish. This thesis investigated how Fundulus killifish cope with daily cycles between hypoxia and normoxia (intermittent hypoxia) by modifying oxygen transport, metabolism, and oxidative stress defense systems. I found that killifish rely on a unique and effective physiological strategy to cope with intermittent hypoxia, and that this strategy is distinct from how they respond to a single bout of hypoxia (followed by normoxia) and to a constant pattern of only hypoxia. This is the most extensive investigation to date on how fish respond to the challenges of intermittent hypoxia, an understudied but ecologically important type of aquatic hypoxia.