Oxidative stress in northern elephant seals: Integration of omics approaches with ecological and experimental studies.

Northern elephant seals experience conditions that increase oxidative stress (OS), including extended fasting, ischemia and hypoxia during breath-holds, and immune responses during colonial breeding. Increased OS is suggested by increases in tissue and plasma concentrations of pro-oxidant enzymes NADPH oxidase and xanthine oxidase (XO). Serum cortisol concentrations were positively associated with XO concentrations and damage markers. Elephant seals exhibit robust anti-oxidant responses as evidenced by increases in anti-oxidant enzymes in plasma and tissues. These responses were sufficient to prevent oxidative damage during breath-holds and extended fasts in juveniles. However, high rates of energy expenditure during breeding were associated with increased evidence for oxidative damage to lipids, proteins and DNA in adults. We integrated investigations of the fasting metabolome and muscle and blubber transcriptomes into our oxidative stress studies. Non-targeted metabolomics analysis of fasting seals identified 227 known metabolites in plasma, including those related to glutathione and purine metabolism. Changes in plasma metabolites suggested that glutathione biosynthesis increased during fasting in weaned pups but not in lactating females. We produced the first reference sequence for elephant seals by RNA sequencing of skeletal muscle and adipose tissue transcriptomes and de novo transcriptome assembly. We annotated muscle and adipose transcripts and identified thousands of genes, including potential mediators of OS. This resource provides elephant seal-specific gene sequences, complements existing metabolite and protein expression studies and provides tools for examining cellular responses to OS in a variety of contexts. We examined changes in tissue gene expression in response to experimental elevation of plasma cortisol. Responses included downregulation of Negative Regulator of Reactive Oxygen Species (NRROS) in muscle, a regulator that limits reactive oxygen species production by tissues. These tools provide novel views of the cellular and systemic mechanisms that enable seals to tolerate high levels of OS.