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      Comparison of Integrated Responses to Nonlethal and Lethal Hypothermal Stress in Milkfish ( Chanos chanos): A Proteomics Study

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          Abstract

          Milkfish is an important aquaculture species in Taiwan, and its high mortality during cold snaps in winter usually causes huge economic losses. To understand the effect of hypothermal stress and the corresponding compensatory stress response in milkfish, this study aimed to compare liver and gill protein levels between milkfish exposed to nonlethal (18°C), lethal (16°C), and control (28°C) temperatures. Using a proteomics approach based on two-dimensional electrophoresis and nano-LC-MS/MS analysis, this study identified thirty unique protein spots from milkfish livers and gills for which protein abundance was significantly different between nonlethal, lethal, and control temperature groups. Proteins identified in the liver were classified into three different categories according to their cellular function: (1) anti-oxidative stress, (2) apoptotic pathway, and (3) cytoskeleton. Similarly, proteins identified in the gill were sorted in five different functional categories: (1) cytoskeleton, (2) immune response, (3) protein quality control, (4) energy production, and (5) intracellular homeostasis. Based on functional information derived from the identified proteins, we assumed that different levels of hypothermal stress had a different effect and induced a different cellular response. Upon nonlethal hypothermal stress, the identified proteins were involved in anti-oxidative stress and anti-inflammation pathways, suggesting that milkfish had high levels of oxidative stress in the liver and exhibited inflammation response in the gill. Upon lethal hypothermal stress, however, identified proteins were associated with apoptosis in the liver and regulation of intracellular homeostasis in the gill. The present study provided evidence to illustrate different multi-physiological responses to nonlethal and lethal hypothermal stress in milkfish livers and gills.

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          The multifunctional fish gill: dominant site of gas exchange, osmoregulation, acid-base regulation, and excretion of nitrogenous waste.

          The fish gill is a multipurpose organ that, in addition to providing for aquatic gas exchange, plays dominant roles in osmotic and ionic regulation, acid-base regulation, and excretion of nitrogenous wastes. Thus, despite the fact that all fish groups have functional kidneys, the gill epithelium is the site of many processes that are mediated by renal epithelia in terrestrial vertebrates. Indeed, many of the pathways that mediate these processes in mammalian renal epithelial are expressed in the gill, and many of the extrinsic and intrinsic modulators of these processes are also found in fish endocrine tissues and the gill itself. The basic patterns of gill physiology were outlined over a half century ago, but modern immunological and molecular techniques are bringing new insights into this complicated system. Nevertheless, substantial questions about the evolution of these mechanisms and control remain.
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            Coping with cold: An integrative, multitissue analysis of the transcriptome of a poikilothermic vertebrate.

            How do organisms respond adaptively to environmental stress? Although some gene-specific responses have been explored, others remain to be identified, and there is a very poor understanding of the system-wide integration of response, particularly in complex, multitissue animals. Here, we adopt a transcript screening approach to explore the mechanisms underpinning a major, whole-body phenotypic transition in a vertebrate animal that naturally experiences extreme environmental stress. Carp were exposed to increasing levels of cold, and responses across seven tissues were assessed by using a microarray composed of 13,440 cDNA probes. A large set of unique cDNAs (approximately 3,400) were affected by cold. These cDNAs included an expression signature common to all tissues of 252 up-regulated genes involved in RNA processing, translation initiation, mitochondrial metabolism, proteasomal function, and modification of higher-order structures of lipid membranes and chromosomes. Also identified were large numbers of transcripts with highly tissue-specific patterns of regulation. By unbiased profiling of gene ontologies, we have identified the distinctive functional features of each tissue's response and integrate them into a comprehensive view of the whole-body transition from one strongly adaptive phenotype to another. This approach revealed an expression signature suggestive of atrophy in cooled skeletal muscle. This environmental genomics approach by using a well studied but nongenomic species has identified a range of candidate genes endowing thermotolerance and reveals a previously unrecognized scale and complexity of responses that impacts at the level of cellular and tissue function.
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              Small heat shock proteins HSP27 (HspB1), αB-crystallin (HspB5) and HSP22 (HspB8) as regulators of cell death.

              Hsp27, αB-crystallin and HSP22 are ubiquitous small heat shock proteins (sHsp) whose expression is induced in response to a wide variety of unfavorable physiological and environmental conditions. These sHsp protect cells from otherwise lethal conditions mainly by their involvement in cell death pathways such as necrosis, apoptosis or autophagy. At a molecular level, the mechanisms accounting for sHsp functions in cell death are (1) prevention of denatured proteins aggregation, (2) regulation of caspase activity, (3) regulation of the intracellular redox state, (4) function in actin polymerization and cytoskeleton integrity and (5) proteasome-mediated degradation of selected proteins. In cancer cells, these sHsp are often overexpressed and associated with increased tumorigenicity, cancer cells metastatic potential and resistance to chemotherapy. Altogether, these properties suggest that Hsp27, αB-crystallin and Hsp22 are appropriate targets for modulating cell death pathways. In the present, we briefly review recent reports showing molecular evidence of cell death regulation by these sHsp and co-chaperones. This article is part of a Directed Issue entitled: Small HSPs in physiology and pathology. Copyright © 2012 Elsevier Ltd. All rights reserved.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                22 September 2016
                2016
                : 11
                : 9
                : e0163538
                Affiliations
                [1 ]Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan
                [2 ]Department of Oceanography, National Sun Yat-Sen University, Kaohsiung 804, Taiwan
                [3 ]Tainan Hydraulics Laboratory, National Cheng Kung University, Tainan 709, Taiwan
                [4 ]Department of Biotechnology, Hung Kuang University, Taichung 433, Taiwan
                [5 ]Agricultural Biotechnology Center, National Chung Hsing University, Taichung 402, Taiwan
                Institut National de la Recherche Agronomique, FRANCE
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: CHC CHT CKK WYL THL.

                • Data curation: CHC.

                • Formal analysis: CHC CHT WYL.

                • Investigation: CHC.

                • Methodology: CHC WYL.

                • Resources: WYL THL.

                • Software: WYL THL.

                • Writing – original draft: CHC.

                • Writing – review & editing: CHT CKK WYL THL.

                Article
                PONE-D-16-17986
                10.1371/journal.pone.0163538
                5033585
                27657931
                d8453a02-abf5-45db-85ba-e9021cd08a80
                © 2016 Chang et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 12 May 2016
                : 10 September 2016
                Page count
                Figures: 5, Tables: 2, Pages: 19
                Funding
                This study was supported by grants to THL from the Ministry of Science and Technology (MOST) of Taiwan (NSC 99-2321-B-005-013-MY3 and 104-2321-B-005-004; https://www.most.gov.tw/en/public) and the Taiwan Comprehensive University System (103TCUS03; http://www.tcus.edu.tw/English).
                Categories
                Research Article
                Biology and Life Sciences
                Biochemistry
                Proteins
                Chaperone Proteins
                Biology and Life Sciences
                Zoology
                Fish Biology
                Fish Physiology
                Biology and Life Sciences
                Zoology
                Animal Physiology
                Vertebrate Physiology
                Fish Physiology
                Physical Sciences
                Physics
                Classical Mechanics
                Mechanical Stress
                Thermal Stresses
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Death
                Apoptosis
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cellular Stress Responses
                Heat Shock Response
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cellular Stress Responses
                Biology and Life Sciences
                Zoology
                Animal Anatomy
                Aquatic Respiratory Anatomy
                Gills
                Biology and Life Sciences
                Anatomy
                Respiratory System
                Gills
                Medicine and Health Sciences
                Anatomy
                Respiratory System
                Gills
                Biology and Life Sciences
                Microbiology
                Bacteriology
                Lethality (Bacteriology)
                Custom metadata
                All relevant data are within the paper.

                Uncategorized
                Uncategorized

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