Dataset Information

In-depth proteomic mapping of skin mucus of gilthead sea bream (Sparus aurata). Surrogate markers of chronic stress in fish farming

ABSTRACT: The skin mucus of gilthead sea bream was mapped by 1-DE followed by liquid chromatography coupled to high resolution mass spectrometry using a quadrupole time-of-flight mass analyzer. More than 2000 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, 2-DE 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. Overall, these results indicate that skin mucus is a reliable tissue for alternative or complementary stress phenotyping in fish farming.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Danio Rerio (zebrafish) (brachydanio Rerio)

TISSUE(S): Epithelial Cell, Skin

SUBMITTER: Luz Valero

LAB HEAD: Jaume Pérez-Sánchez

PROVIDER: PXD004115 | Pride | 2018-10-26

REPOSITORIES: Pride

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Publications

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

Pérez-Sánchez Jaume J Terova Genciana G Simó-Mirabet Paula P Rimoldi Simona S Folkedal Ole O Calduch-Giner Josep A JA Olsen Rolf E RE Sitjà-Bobadilla Ariadna A

Frontiers in physiology 20170201

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 ...[more]

PMID: 28210224

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Project description:Antimicrobial peptides (AMPs) constitute a broad range of bioactive compounds in diverse organisms, including fish. They are effector molecules for the innate immune response, against pathogens, tissue damage and infections. Still, AMPs from African Catfish, Clarias gariepinus skin mucus are largely unexplored despite their possible therapeutic role in combating antimicrobial resistance. In this study, African Catfish Antimicrobial peptides (ACAPs) were identified from the skin mucus of African Catfish, C. gariepinus. Native peptides were extracted from fish mucus scrapings in 10% acetic acid (v/v) and ultra-filtered using 5kDa molecular cut-off membrane. The extract was purified using C18 Solid Phase Extraction. The antibacterial activity was determined using the Agar Well Diffusion method and broth-dilution method utilizing Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922). Thereafter, Sephadex G-25 gel filtration was further utilized in bio-guided isolation of the most active fractions prior to peptide identification using Orbitrap Fusion Lumos Tribrid Mass Spectrometry. The skin mucus extracted from African Catfish from all the three major lakes of Uganda exhibited antimicrobial activity on E. coli and S. aureus. Lake Albert’s C. gariepinus demonstrated the best activity with the lowest MIC of 2.84 µg/mL and 0.71 µg/mL on S. aureus and E. coli respectively. Sephadex G-25 peak I mass spectrometry analysis alongside in silico analysis revealed seven short peptides (11-15 amino acid residues) of high antimicrobial scores (0.561-0.905 units). In addition, these peptides had a low molecular weight (1005.57-1622.05 Da), and had percentage hydrophobicity above 54%. Up to four of these antimicrobial peptides demonstrated α-helix structure conformation, rendering them amphipathic. The ﬁndings of this study indicate that novel antimicrobial peptides can be sourced from the skin mucus of C. gariepinus. Such antimicrobial peptides are potential alternatives to the traditional antibiotics and can be of great application to food and pharmaceutical industries; however, further studies are still needed to establish their drug-likeness and safety profiles.

Project description:Animal mucosal barriers constantly interact with the external environment and this interaction is markedly different in aquatic and terrestrial environments. Transitioning from water to land was a critical step in vertebrate evolution but the immune adaptations that mucosal barriers such as the skin underwent during that process are essentially unknown. Vertebrate animals such as the African lungfish have a bimodal life, switching from freshwater to terrestrial habitats when environmental conditions are not favorable. African lungfish skin mucus secretions contribute to the terrestrialization process by forming a cocoon that surrounds and protect the lungfish body. The goal of this study is to characterize the skin mucus immunoproteome of African lungfish, Protopterus dolloi, before and during the induction phase of terrestrialization as well as the immunoproteome of the gill mucus during the terrestrialization induction phase. Using LC-MS/MS, we identified a total of a total of 974 proteins using a lungfish Illumina RNA-seq database and 880 proteins using a lungfish 454 RNA-seq database for annotation in the three samples analyzed (control skin mucus, terrestrialized skin mucus and terrestrialized gill mucus). The terrestrialized skin mucus proteome was enriched in proteins with known antimicrobial functions such as histones and S100 proteins. In support, gene ontology analyses showed that the terrestrialized skin mucus proteome has predicted functions in processes such as viral process, defense response to Gram negative bacterium and tumor necrosis factor mediated signaling. Importantly, we observed a switch in immunoglobulin heavy chain secretion upon terrestrialization, with IgW1L and IgM1 present in control skin mucus and IgW1L, IgM1 and IgM2 in terrestrialized skin mucus. Combined, these results indicate an increase investment in the production of unique immune molecules in P. dolloi skin mucus in response to terrestrialization that likely better protect lungfish against external aggressors found in land.

Dataset Information

In-depth proteomic mapping of skin mucus of gilthead sea bream (Sparus aurata). Surrogate markers of chronic stress in fish farming

Publications

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

Similar Datasets

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