Changes in Liver Proteome Expression of Senegalese Sole (Solea senegalensis) in Response to Repeated Handling Stress

Abstract

The Senegalese sole, a high-value flatfish, is a good candidate for aquaculture production. Nevertheless, there are still issues regarding this species' sensitivity to stress in captivity. We aimed to characterize the hepatic proteome expression for this species in response to repeated handling and identify potential molecular markers that indicate a physiological response to chronic stress. Two groups of fish were reared in duplicate for 28 days, one of them weekly exposed to handling stress (including hypoxia) for 3 min, and the other left undisturbed. Two-dimensional electrophoresis enabled the detection of 287 spots significantly affected by repeated handling stress (Wilcoxon-Mann-Whitney U test, p < 0.05), 33 of which could be reliably identified by peptide mass spectrometry. Chronic exposure to stress seems to have affected protein synthesis, folding and turnover (40S ribosomal protein S12, cathepsin B, disulfide-isomerase A3 precursor, cell-division cycle 48, and five distinct heat shock proteins), amino acid metabolism, urea cycle and methylation/folate pathways (methionine adenosyltransferase I α, phenylalanine hydroxylase, mitochondrial agmatinase, serine hydroxymethyltransferase, 3-hydroxyanthranilate 3,4-dioxygenase, and betaine homocysteine methyltransferase), cytoskeletal (40S ribosomal protein SA, α-actin, β-actin, α-tubulin, and cytokeratin K18), aldehyde detoxification (aldehyde dehydrogenase 4A1 family and aldehyde dehydrogenase 7A1 family), carbohydrate metabolism and energy homeostasis (fatty acid-binding protein, enolase 3, enolase 1, phosphoglycerate kinase, glyceraldehyde-3-phosphate dehydrogenase, aconitase 1, mitochondrial ATP synthase α-subunit, and electron-transfer flavoprotein α polypeptide), iron and selenium homeostasis (transferrin and selenium binding protein 1), steroid hormone metabolism (3-oxo-5-β-steroid 4-dehydrogenase), and purine salvage (hypoxanthine phosphoribosyltransferase). Further characterization is required to fully assess the potential of these markers for the monitoring of fish stress response to chronic stressors of aquaculture environment.

Proteomic analysis of anatoxin-a acute toxicity in zebrafish reveals gender specific responses and additional mechanisms of cell stress

“…The contrasting abundances of the two muscle eno1 isoforms may indicate a turn-over of activity controlled by post-translational modification. Eno1 has been regarded as a marker of stress and disease (Braceland et al, 2013;Cordeiro et al, 2012;Wozny et al, 2012). Eno1 has been also considered an important neurotrophic factor and is under the transcriptional regulation of a number of stimuli, such as ischemia, hypoxia, cytokines and mitogens (Li et al, 2014;Sousa et al, 2005).…”

PROTEOMICS in aquaculture: Applications and trends

“…Liver tissue also displays energy metabolism shifts in response to stress, but at a much deeper level, reflecting its essential role in physiological detoxification and energy management: adaptations usually cover amino acid and lipid metabolism, besides carbohydrate metabolism. Other often affected pathways in liver include the urea cycle, the methionine/homocysteine cycle, the folate one-carbon pool and betaine metabolism, which are intimately connected to amino acid metabolism and detoxification processes [29,57,179]. Besides these main pathways, cellular stress also induces changes in the expression of proteins related to signaling, cell cycle regulation, transcription factors, ribosomal proteins and others.…”

“…The study of the impact of xenobiotics in aquaculture has also benefited from proteomics technologies, with several studies done on the impact of biotoxins [141,[151][152][153], pharmaceuticals [58,154,155], hormones [15,46,156], metals [157,158] and other pollutants [43,45,67,[159][160][161][162][163] on the proteomes of aquaculture aquatic organisms. Finally, there are also some studies correlating environmental sources of stress in aquaculture (hypoxia [57], anoxia [26,[164][165][166], hyperoxygenation [167], osmotic [168][169][170][171][172][173] and temperature [174][175][176][177], as well as stress induced by management practices (like high stocking densities [29,178], handling [29,179] and pre-slaughter stress [180]), with proteome changes in several tissues.…”

Proteomic evaluation of potentiated sulfa treatment on gilthead sea bream (Sparus aurata L.) liver

“…The increase in the levels of ALDH2 found in treated fish could reflect an increase in their capacity to process aldehydes and, therefore to protect the liver from oxidative stress. In fact, a down regulation of ALDHs enzymes in fish liver has been related to a diminution of the capacity to protect this organ from oxidative stress (Cordeiro et al 2012), and also with a non-induction of detoxification processes (Rufino-Palomares et al 2011 (Eze 1983;Gaudet et al 1975). The enzyme is found in the liver, heart, skeletal muscle, kidneys, brain, and red blood cells, and changes in aminotransferases activities are frequently used as biomarker to determine fish liver damage (De la Torre et al 2005;Prashanth and Neelagund 2008;Inyang et al 2010).…”

“…However, the increase in the level of liver FABP found in gilthead sea bream and Senegalese sole subjected to handling and crowing stress, was associated with an increase in lipid mobilization, in order to respond to the increased energetic requirements due to stress, as well as in preventing oxidative damage (Alves et al 2010;Cordeiro et al 2012). …”