Hypoxic condition is a naturally occurring environmental stressor in aquatic ecosystems. However, due to modern anthropocentric activities, hypoxia has been increasing in prevalence and severity. Rainbow trout, a keystone species in many North American lakes, is hypoxia intolerant. As a result, this species is of particular concern when studying the effects of hypoxia on an organism’s physiological functioning. Chronic starvation was used as a tool to induce metabolic depression to determine the effect that depressed metabolic rate had on hypoxia tolerance. Juvenile rainbow trout were deprived of food for five weeks at 15oC. Each week, routine metabolic rate (RMR) and critical oxygen tension (Pcrit) were measured. Concomitantly, resting and post-hypoxia fish (8 h at ~50% air saturation) were sampled to measure metabolites in blood, liver and muscle, as well as enzyme activities in select tissues. Food deprivation resulted in a decrease in routine metabolic rate (RMR) and shift towards an increased reliance on aerobic metabolism. Pcrit decreased significantly following four weeks of food deprivation respectively, indicating that metabolic depression induced by food deprivation may confer an increased tolerance to low environmental oxygen concentration ([O2]). However, marginal metabolic scope (MMS), another indicator of hypoxia tolerance, did not change in response to metabolic depression. Furthermore, subjecting trout to O2 limitation resulted in mobilization of carbohydrates from the liver subsequently leading to hyperglycemia. This was likely a survival technique ensuring that if severe hypoxia ensues, anaerobic substrates are ready for transport to the necessary tissues. / Thesis (Master, Biology) -- Queen's University, 2011-10-12 23:21:04.517

This research examined the effects of acclimation to lowered salinity, elevated temperature, and hypoxia on aerobic metabolism of the Gulf killifish, Fundulus grandis, a common estuarine resident of the Gulf of Mexico. Standard metabolic rate (SMR), maximum metabolic rate (MMR), absolute aerobic scope (AAS), and critical oxygen tension (Pcrit) were each influenced by one or more acclimation treatments. Assessing the consistency of these traits measured in the same individuals over time, all were found to be significantly repeatable with no indication that the repeatability of any traits was affected by acclimation conditions. Significant correlations were found between SMR and Pcrit (positively correlated), between SMR and AAS (negatively correlated), between MMR and AAS (positive), and between AAS and Pcrit (negative). This study, therefore, documents the effects of acclimation on these traits, their repeatability, and correlations among them. It further suggests that repeatability of these traits is not context dependent.

Marine mammals such as northern elephant seals (NES) routinely experience hypoxemia and ischemia-reperfusion events to many tissues during deep dives with no apparent adverse effects. Adaptations to diving include increased antioxidants and elevated oxygen storage capacity associated with high hemoprotein content in blood and muscle. Despite experiencing decreased oxygen tensions during diving, NES likely rely on the mobilization of large lipids stores and catabolism of fatty acids to provide energy to exercising muscle while diving. To identify potential regulatory mechanisms that may underly hypoxia and exercise tolerance in diving mammals, this study used system-wide approaches to characterize changes in genes and proteins in two metabolically active tissues (skeletal muscle and blubber) and whole blood of NES over development and in response to translocation. Specifically, this study profiled muscle and blood gene expression associated with regulation of oxidative stress and inflammatory pathways in weaned pups, juveniles, and adult NES as well as evaluated muscle and blubber transcriptomic and proteomic responses to swimming and diving in juvenile NES. I found that expression of genes associated with mitochondrial biogenesis (PGC1A, ESRRA, ESRRG), immune system activation (HMOX2, IL1B, NRF2, BVR, IL10), and protection from lipid peroxidation (GPX4, PRDX6, PRDX1, SIRT1) increased over postnatal development in muscle and whole blood of NES, providing a potential ontogenic mechanism for increasing diving capacity and hypoxia and ischemia-reperfusion tolerance. I also found that expression of genes and abundance of proteins associated with lipid transport (APOD, ABCA6, ABCA8, ABCA10, CD1E), lipid catabolism (ADIPOQ , ENPP6), and adipogenesis (DLK1, ADIRF,) increased, while those associated with insulin sensitivity and energy expenditure (APLN, VGF) decreased in response to swimming and diving in juvenile NES blubber and muscle, suggesting potential mechanisms for fuel provisioning to muscle during exercise in hypoxic conditions. Together, these data provide insights into gene activity in muscle, blubber, and blood cells that may provide hypoxia tolerance and regulate energy homeostasis and exercise performance during breath holds in diving mammals.

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.