SUMMARYAntarctic icefishes have a significantly lower critical thermal maximum (CT max ) compared with most red-blooded notothenioid fishes. We hypothesized that the lower thermal tolerance of icefishes compared with red-blooded notothenioids may stem from a greater vulnerability to oxidative stress as temperature increases. Oxidative muscles of icefishes have high volume densities of mitochondria, rich in polyunsaturated fatty acids, which can promote the production of reactive oxygen species (ROS). Moreover, icefishes have lower levels of antioxidants compared with red-blooded species. To test our hypothesis, we measured levels of oxidized proteins and lipids, and transcript levels and maximal activities of antioxidants in heart ventricle and oxidative pectoral adductor muscle of icefishes and red-blooded notothenioids held at 0°C and exposed to their CT max . Levels of oxidized proteins and lipids increased in heart ventricle of some icefishes but not in red-blooded species in response to warming, and not in pectoral adductor muscle of any species. Thus, increases in oxidative damage in heart ventricles may contribute to the reduced thermal tolerance of icefishes. Despite an increase in oxidative damage in hearts of icefishes, neither transcript levels nor activities of antioxidants increased, nor did they increase in any tissue of any species in response to exposure to CT max . Rather, transcript levels of the enzyme superoxide dismutase (SOD) decreased in hearts of icefishes and the activity of SOD decreased in hearts of the red-blooded species Gobionotothen gibberifrons. These data suggest that notothenioids may have lost the ability to elevate levels of antioxidants in response to heat stress.

Understanding what limits the capacity of organisms to tolerate increasing temperatures is a critical objective in comparative biology. Using an experimental system of Antarctic notothenioid fishes, we sought to determine whether a mismatch between oxygen demand and oxygen supply was responsible for setting thermal tolerance limits. Previous studies have shown that Antarctic icefishes (family Channichthyidae), which lack hemoglobin, have lower critical thermal maxima (CTmax) than red-blooded notothenioids collected from the same region of the Antarctic (Western Antarctic Peninsula). In addition, within the notothenioid fishes there exists a positive correlation between CTmax and hematocrit. We tested the hypothesis that the lower CTmax of icefishes is associated with reduced oxygen supply. We employed an experimental heat ramp (4°C h(-1)) to determine CTmax under both normoxic and hyperoxic conditions and quantified correlates of oxygen limitation (lactate levels and expression of hypoxia-inducible factor-1α) in white-blooded Chaenocephalus aceratus and red-blooded Notothenia coriiceps. Hyperoxia, corresponding to a three- to fourfold increase in seawater Po2, did not extend CTmax in either species despite an overall mitigation in the rise of plasma and muscle lactate compared with the normoxic treatment. Our results also indicate that cardiac HIF-1α mRNA levels were insensitive to changes in both temperature and oxygen treatments. The absence of a change in CTmax with hyperoxia is likely to represent the contribution of factors beyond oxygen supply to physiological failure at elevated temperatures.