Skin Mucus of Gilthead Sea Bream (Sparus aurata L.). Protein Mapping and Regulation in Chronically Stressed Fish

Abstract

The skin mucus of gilthead sea bream was mapped by one-dimensional gel electrophoresis followed by liquid chromatography coupled to high resolution mass spectrometry using a quadrupole time-of-flight mass analyzer. More than 2,000 proteins were identified with a protein score filter of 30. The identified proteins were represented in 418 canonical pathways of the Ingenuity Pathway software. After filtering by canonical pathway overlapping, the retained proteins were clustered in three groups. The mitochondrial cluster contained 59 proteins related to oxidative phosphorylation and mitochondrial dysfunction. The second cluster contained 79 proteins related to antigen presentation and protein ubiquitination pathways. The third cluster contained 257 proteins where proteins related to protein synthesis, cellular assembly, and epithelial integrity were over-represented. The latter group also included acute phase response signaling. In parallel, two-dimensional gel electrophoresis methodology identified six proteins spots of different protein abundance when comparing unstressed fish with chronically stressed fish in an experimental model that mimicked daily farming activities. The major changes were associated with a higher abundance of cytokeratin 8 in the skin mucus proteome of stressed fish, which was confirmed by immunoblotting. Thus, the increased abundance of markers of skin epithelial turnover results in a promising indicator of chronic stress in fish.

Protein changes as robust signatures of fish chronic stress: a proteomics approach to fish welfare research

“…Proteins considered to be "positive" APP were likewise shown to be increased in abundance in the plasma of fish stressed by net handling (fibrinogen alpha-chain, complement component C3, haptoglobin, complement factor B, warm-temperature acclimation 65 kDa protein, alpha-1-antitrypsin), while proteins considered as "negative" were decreased (transferrin, interalpha-trypsin inhibitor, apolipoprotein A-I) [70]. A diverse number of proteins involved in the APR was also found previously to be modulated in chronically stressed gilthead seabream [71].…”

Biological and Ecological Roles of External Fish Mucus: A Review

“…The use of high-throughput techniques such as proteomics allows for the pursuit of comprehensive comparative studies to better understand fish mucus dynamics. Proteomics studies have cast light on fish mucus dynamics of infected fish [181][182][183], overcrowding [184], chronic stress [185], and fish exposed to different diets [186].…”

Chronic wounds alter the proteome profile in skin mucus of farmed gilthead seabream

“…One of the limitations of this approach could be the limited range of molecular weights available, thus mucins and other high molecular weight proteins have been undetected in these works. A recently published proteome map of gilthead seabream with more than 2000 proteins used 1-DE gels and mass spectrometry and any mucin was identified [ 40 ].…”