Activation of oxygen-responsive pathways are associated with altered protein metabolism in Arctic char exposed to hypoxia

Abstract

Fish exposed to fluctuating oxygen concentrations often alter their metabolism and/or behaviour to survive. Hypoxia tolerance is typically associated with the ability to reduce energy demand by supressing metabolic processes such as protein synthesis. Arctic char is amongst the most sensitive salmonid to hypoxia, and typically engage in avoidance behaviour when faced with lack of oxygen. We hypothesized that a sensitive species will still have the ability (albeit reduced) to regulate molecular mechanisms during hypoxia. We investigated the tissue-specific response of protein metabolism during hypoxia. Little is known on protein degradation pathways during hypoxia in fish and we predict that protein degradation pathways are differentially regulated and play a role in the hypoxia response. We also studied the regulation of oxygen-responsive cellular signalling pathways (Hypoxia inducible factor, unfolded protein response and mTOR pathway) since most of what we know comes from studies on cancerous mammalian cell lines. Arctic char, were exposed to a cumulative, graded hypoxia trials, for 3 hours at each air saturation level (100%, 50%, 30% and 15%). The rate of protein synthesis was measured using a flooding dose technique, while protein degradation and signalling pathways were assessed by measuring transcripts and phosphorylation of target proteins. Protein synthesis decreased in all tissues measured (liver, muscle, gill, digestive system) except for the heart. Salmonid hearts have preferential access to oxygen through a well-developed coronary artery, therefore the heart is likely the last tissue to become hypoxic. Autophagy markers were upregulated in the liver, while protein degradation markers were downregulated in the heart during hypoxia. Further work is needed to determine the effects of a decrease in protein degradation on a hypoxic salmonid heart. Our study showed that protein metabolism in Arctic char is altered in a tissue-specific fashion during graded hypoxia, which is in accordance with the responses of the three major hypoxia-sensitive pathways (HIF, UPR and mTOR). The activation pattern of these pathways and the cellular processes that are under their control varies greatly among tissues, sometimes even going in opposite direction. This study provides new insights on the effects of hypoxia on protein metabolism. The adjustments of these cellular processes likely contribute in shifting the fish phenotype into a more hypoxia tolerant one, if more than one hypoxia event were to occur. Our results warrant studying these adjustments in fish exposed to long-term and diel cycling hypoxia.