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

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          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.

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          Barrier properties of mucus.

          Mucus is tenacious. It sticks to most particles, preventing their penetration to the epithelial surface. Multiple low-affinity hydrophobic interactions play a major role in these adhesive interactions. Mucus gel is also shear-thinning, making it an excellent lubricant that ensures an unstirred layer of mucus remains adherent to the epithelial surface. Thus nanoparticles (NP) must diffuse readily through the unstirred adherent layer if they are to contact epithelial cells efficiently. This article reviews some of the physiological and biochemical properties that form the mucus barrier. Capsid viruses can diffuse through mucus as rapidly as through water and thereby penetrate to the epithelium even though they have to diffuse 'upstream' through mucus that is being continuously secreted. These viruses are smaller than the mucus mesh spacing, and have surfaces that do not stick to mucus. They form a useful model for developing NP for mucosal drug delivery.
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            Peroxiredoxins: guardians against oxidative stress and modulators of peroxide signaling.

            Peroxiredoxins (Prxs) are a ubiquitous family of cysteine-dependent peroxidase enzymes that play dominant roles in regulating peroxide levels within cells. These enzymes, often present at high levels and capable of rapidly clearing peroxides, display a remarkable array of variations in their oligomeric states and susceptibility to regulation by hyperoxidative inactivation and other post-translational modifications. Key conserved residues within the active site promote catalysis by stabilizing the transition state required for transferring the terminal oxygen of hydroperoxides to the active site (peroxidatic) cysteine residue. Extensive investigations continue to expand our understanding of the scope of their importance as well as the structures and forces at play within these critical defense and regulatory enzymes.
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              An Overview of the Immunological Defenses in Fish Skin

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                Author and article information

                Contributors
                Journal
                Front Physiol
                Front Physiol
                Front. Physiol.
                Frontiers in Physiology
                Frontiers Media S.A.
                1664-042X
                01 February 2017
                2017
                : 8
                : 34
                Affiliations
                [1] 1Nutrigenomics and Fish Growth Endocrinology Group, Biology, Culture and Pathology of Marine Species, Institute of Aquaculture Torre de la Sal (IATS-CSIC) Castellón, Spain
                [2] 2Department of Biotechnology and Life Sciences, University of Insubria Varese, Italy
                [3] 3Inter-University Centre for Research in Protein Biotechnologies “The Protein Factory” Polytechnic University of Milan and University of Insubria Varese, Italy
                [4] 4Institute of Marine Research Matre Matredal, Norway
                [5] 5Department of Biology, Norwegian University for Science and Technology Trondheim, Norway
                [6] 6Fish Pathology Group Group, Biology, Culture and Pathology of Marine Species, Institute of Aquaculture Torre de la Sal (IATS-CSIC) Castellón, Spain
                Author notes

                Edited by: Pung P. Hwang, Academia Sinica, Taiwan

                Reviewed by: Alberto Cuesta, University of Murcia, Spain; Mathilakath Vijayan, University of Calgary, Canada

                *Correspondence: Jaume Pérez-Sánchez jaime.perez.sanchez@ 123456csic.es

                This article was submitted to Aquatic Physiology, a section of the journal Frontiers in Physiology

                Article
                10.3389/fphys.2017.00034
                5288811
                0ea396c3-ea31-4cba-8b21-5fee26d23398
                Copyright © 2017 Pérez-Sánchez, Terova, Simó-Mirabet, Rimoldi, Folkedal, Calduch-Giner, Olsen and Sitjà-Bobadilla.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 05 October 2016
                : 13 January 2017
                Page count
                Figures: 2, Tables: 4, Equations: 0, References: 81, Pages: 18, Words: 11295
                Funding
                Funded by: Seventh Framework Programme 10.13039/501100004963
                Award ID: 262336
                Funded by: Ministerio de Economía y Competitividad 10.13039/501100003329
                Award ID: AGL2013-48560
                Funded by: Generalitat Valenciana 10.13039/501100003359
                Award ID: PROMETEO FASE II-2014/085
                Categories
                Physiology
                Original Research

                Anatomy & Physiology
                chronic stress,cytokeratins,gilthead sea bream,proteome,skin mucus
                Anatomy & Physiology
                chronic stress, cytokeratins, gilthead sea bream, proteome, skin mucus

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