Warming-induced "plastic floors" improve hypoxia vulnerability, not aerobic scope, in red drum (Sciaenops ocellatus).

Ocean warming is a prevailing threat to marine ectotherms. Recently the "plastic floors, concrete ceilings" hypothesis was proposed, which suggests that a warmed fish will acclimate to higher temperatures by reducing standard metabolic rate (SMR) while keeping maximum metabolic rate (MMR) stable, therefore improving aerobic scope (AS). Here we evaluated this hypothesis on red drum (Sciaenops ocellatus) while incorporating measures of hypoxia vulnerability (critical oxygen threshold; Pcrit) and mitochondrial performance. Fish were subjected to a 12-week acclimation to 20 °C or 28 °C. Respirometry was performed every 4 weeks to obtain metabolic rate and Pcrit; mitochondrial respirometry was performed on liver and heart samples at the end of the acclimation. 28 °C fish had a significantly higher SMR, MMR, and Pcrit than 20 °C controls at time 0, but SMR declined by 36.2 % over the 12-week acclimation. No change in SMR was observed in the control treatment. Contrary to expectations, SMR suppression did not improve AS relative to time 0 owing to a progressive decline in MMR over acclimation time. Pcrit decreased by 27.2 % in the warm-acclimated fishes, which resulted in temperature treatments having statistically similar values by 12-weeks. No differences in mitochondrial traits were observed in the heart - despite a Δ8 °C assay temperature - while liver respiratory and coupling control ratios were significantly improved, suggesting that mitochondrial plasticity may contribute to the reduced SMR with warming. Overall, this work suggests that warming induced metabolic suppression offsets the deleterious consequences of high oxygen demand on hypoxia vulnerability, and in so doing greatly expands the theoretical range of metabolically available habitats for red drum.

Hypoxia acclimation improves mitochondrial efficiency in the aerobic swimming muscle of red drum (Sciaenops ocellatus).

Environmental hypoxia (low dissolved oxygen) is a significant threat facing fishes. As fishes require oxygen to efficiently produce ATP, hypoxia can significantly limit aerobic capacity. However, some fishes show respiratory flexibility that rescues aerobic performance, including plasticity in mitochondrial performance. This plasticity may result in increased mitochondrial efficiency (e.g., less proton leak), increased oxygen storage capacity (increased myoglobin), and oxidative capacity (e.g., higher citrate synthase activity) under hypoxia. We acclimated a hypoxia-tolerant fish, red drum (Sciaenops ocellatus), to 8-days of constant hypoxia to induce a hypoxic phenotype. Fish were terminally sampled for cardiac and red muscle tissue to quantify oxidative phosphorylation, proton leak, and maximum respiration in tissue from both hypoxia-acclimated and control fish. Tissue was also collected to assess the plasticity of citrate synthase enzyme activity and mRNA expression for select oxygen storage and antioxidant pathway transcripts. We found that mitochondrial respiration rates were not affected by hypoxia exposure in cardiac tissue, though citrate synthase activity and myoglobin expression were higher following hypoxia acclimation. Interestingly, measures of mitochondrial efficiency in red muscle significantly improved in hypoxia-acclimated individuals. Hypoxia-acclimated fish had significantly higher OXPHOS Control Efficiency, OXPHOS Capacity and Coupling Control Ratios (i.e., LEAK/OXPHOS). There was no significant change to citrate synthase activity or myoglobin expression in red muscle. Overall, these results suggest that red muscle mitochondria of hypoxia-acclimated fish more efficiently utilize oxygen, which may explain previous reports in red drum of improved aerobic swimming performance in the absence of improved maximum metabolic rate following hypoxia acclimation.

The effects of size on exhaustive exercise and recovery in a marine sportfish, the red drum (Sciaenops ocellatus).

Recreational angling is an economically important activity in many communities around the world. One conservation strategy adopted to offset the population-level consequences of recreational angling is "catch-and-release" (CAR), which is the act of returning fish to the environment following an angling event. While an expansive literature has helped to generalize CAR best practices, species-specific validation of recovery profiles remains a crucial component of species-specific angling guidance. This study sought to define the injury and recovery profiles in the plasma and white muscle following exhaustive exercise in two size classes of a common Gulf of Mexico sportfish, the red drum (Sciaenops ocellatus). The two sizes included a "small" (20-30 cm) and "slot" size (51-74 cm), the latter of which is a common angling target. Both size classes showed a characteristic injury profile that consisted of significantly elevated muscle and plasma lactate, plasma osmolality and haematocrit, as well as decreased muscle ATP and phosphocreatine, and lowered plasma and muscle pH. In small fish, muscle metabolites returned to control values by 1 h post-exercise and plasma metabolites returned to control between 3 and 6 h post-exercise. In contrast, slot sized fish had recovery periods of ≥3 h for all metabolites. The maximum injury effect size was also greater in the slot size class. These data suggest that while red drum conform to typical patterns of post-exercise recovery, larger trophy-sized fish may be more at risk to the ancillary effects of exhaustive exercise owing to greater exercise injury and slower recovery rates.

The role of carbonic anhydrase-mediated tissue oxygen extraction in a marine teleost acclimated to hypoxia.

With the growing prevalence of hypoxia (O2 levels ≤2 mg l-1) in aquatic and marine ecosystems, there is increasing interest in the adaptive mechanisms fish may employ to better their performance in stressful environments. Here, we investigated the contribution of a proposed strategy for enhancing tissue O2 extraction - plasma-accessible carbonic anhydrase (CA-IV) - under hypoxia in a species of estuarine fish (red drum, Sciaenops ocellatus) that thrives in fluctuating habitats. We predicted that hypoxia-acclimated fish would increase the prevalence of CA-IV in aerobically demanding tissues to confer more efficient tissue O2 extraction. Furthermore, we predicted the phenotypic changes to tissue O2 extraction that occur with hypoxia acclimation may improve respiratory and swim performance under 100% O2 conditions (i.e. normoxia) when compared with performance in fish that have not been acclimated to hypoxia. Interestingly, there were no significant differences in relative CA-IV mRNA expression, protein abundance or enzyme activity between the two treatments, suggesting CA-IV function is maintained under hypoxia. Likewise, respiratory performance of hypoxia-acclimated fish was similar to that of control fish when tested in normoxia. Critical swim speed (Ucrit) was significantly higher in hypoxia-acclimated fish but translated to marginal ecological benefits with an increase of ∼0.3 body lengths per second. Instead, hypoxia-acclimated fish may have relied more heavily on anaerobic metabolism during their swim trials, utilizing burst swimming 1.5 times longer than control fish. While the maintenance of CA-IV may still be an important contributor for hypoxia tolerance, our evidence suggests hypoxia-acclimated red drum are using other mechanisms to cope in an O2-depleted environment.

Respiratory plasticity improves aerobic performance in hypoxia in a marine teleost.

Ocean deoxygenation is a pressing concern in the face of climate change. In response to prolonged hypoxia, fishes have demonstrated the ability to dynamically regulate hemoglobin (Hb) expression to enhance oxygen (O2) uptake. Here, we examined hypoxia-inducible Hb expression in red drum (Sciaenops ocellatus) and the subsequent implications on Hb-O2 binding affinity and aerobic scope. Fish were acclimated to 30 % air saturation for 1 d, 4 d, 8 d, 2 w, or 6 w, and red blood cells were collected for gene expression and biochemical profiling. Hypoxia acclimation induced significant up-regulation of one Hb subunit isoform (hbα 2) relative to control by 4 d with consistent upregulation thereafter. Hematocrit increased in hypoxia, with no changes in the allosteric modulator [NTP] at any time point. Changes in Hb expression co-occurred with a reduced Root effect (~26 % in normoxia, ~14 % in hypoxia) at a physiologically relevant pH while increasing O2 binding affinity (i.e., lower P50). These changes correlated with increased maximum metabolic rate and aerobic scope relative to controls when fish were tested in hypoxia. These results demonstrate an important role for Hb multiplicity in improving O2 affinity and maximizing respiratory performance in hypoxia.

Is hypoxia vulnerability in fishes a by-product of maximum metabolic rate?

The metabolic index concept combines metabolic data and known thermal sensitivities to estimate the factorial aerobic scope of animals in different habitats, which is valuable for understanding the metabolic demands that constrain species' geographical distributions. An important assumption of this concept is that the O2 supply capacity (which is equivalent to the rate of oxygen consumption divided by the environmental partial pressure of oxygen: ) is constant at O2 tensions above the critical O2 threshold (i.e. the where O2 uptake can no longer meet metabolic demand). This has led to the notion that hypoxia vulnerability is not a selected trait, but a by-product of selection on maximum metabolic rate. In this Commentary, we explore whether this fundamental assumption is supported among fishes. We provide evidence that O2 supply capacity is not constant in all fishes, with some species exhibiting an elevated O2 supply capacity in hypoxic environments. We further discuss the divergent selective pressures on hypoxia- and exercise-based cardiorespiratory adaptations in fishes, while also considering the implications of a hypoxia-optimized O2 supply capacity for the metabolic index concept.

A methodological evaluation of the determination of critical oxygen threshold in an estuarine teleost.

One measure of hypoxia tolerance is the critical oxygen threshold, Pcrit, which is the point where standard metabolism can no longer be maintained through aerobic processes. Traditionally, Pcrit was determined using closed respirometry, whereby the fish's respiration naturally lowered O2 More recently, intermittent flow techniques have been adopted, where N2 is used to displace O2, which ostensibly reduces end-product build-up. This study used a paired design on the marine teleost, red drum. Pcrit is comparable between closed (4.6±0.2 kPa; mean±s.e.m.) and intermittent flow (4.4±0.2 kPa; mean±s.e.m.) respirometry. pCO2, ammonia and pH changes within the chamber were measured prior to the onset of Pcrit and at the end of a typical Pcrit trial and revealed changes in water chemistry in both closed and intermittent flow. Pcrit values were similar in both methods of hypoxia induction regardless of subsequent water chemistry changes that occurred in both methods.