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      Transcriptome Analysis of the Hepatopancreas in the Pacific White Shrimp ( Litopenaeus vannamei) under Acute Ammonia Stress

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          Abstract

          In the practical farming of Litopenaeus vannamei, the intensive culture system and environmental pollution usually results in a high concentration of ammonia, which usually brings large detrimental effects to shrimp, such as increasing the susceptibility to pathogens, reducing growth, decreasing osmoregulatory capacity, increasing the molting frequency, and even causing high mortality. However, little information is available on the molecular mechanisms of the detrimental effects of ammonia stress in shrimp. In this study, we performed comparative transcriptome analysis between ammonia-challenged and control groups from the same family of L. vannamei to identify the key genes and pathways response to ammonia stress. The comparative transcriptome analysis identified 136 significantly differentially expressed genes that have high homologies with the known proteins in aquatic species, among which 94 genes are reported potentially related to immune function, and the rest of the genes are involved in apoptosis, growth, molting, and osmoregulation. Fourteen GO terms and 6 KEGG pathways were identified to be significantly changed by ammonia stress. In these GO terms, 13 genes have been studied in aquatic species, and 11 of them were reported potentially involved in immune defense and two genes were related to molting. In the significantly changed KEGG pathways, all the 7 significantly changed genes have been reported in shrimp, and four of them were potentially involved in immune defense and the other three were related to molting, defending toxicity, and osmoregulation, respectively. In addition, majority of the significantly changed genes involved in nitrogen metabolisms that play an important role in reducing ammonia toxicity failed to perform the protection function. The present results have supplied molecular level support for the previous founding of the detrimental effects of ammonia stress in shrimp, which is a prerequisite for better understanding the molecular mechanism of the immunosuppression from ammonia stress.

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          Most cited references48

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          Heat-shock protein 70 inhibits apoptosis by preventing recruitment of procaspase-9 to the Apaf-1 apoptosome.

          The cellular-stress response can mediate cellular protection through expression of heat-shock protein (Hsp) 70, which can interfere with the process of apoptotic cell death. Stress-induced apoptosis proceeds through a defined biochemical process that involves cytochrome c, Apaf-1 and caspase proteases. Here we show, using a cell-free system, that Hsp70 prevents cytochrome c/dATP-mediated caspase activation, but allows the formation of Apaf-1 oligomers. Hsp70 binds to Apaf-1 but not to procaspase-9, and prevents recruitment of caspases to the apoptosome complex. Hsp70 therefore suppresses apoptosis by directly associating with Apaf-1 and blocking the assembly of a functional apoptosome.
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            Prediction of potential GPI-modification sites in proprotein sequences.

            Glycosylphosphatidylinositol (GPI) lipid anchoring is a common posttranslational modification known mainly from extracellular eukaryotic proteins. Attachment of the GPI moiety to the carboxyl terminus (omega-site) of the polypeptide follows after proteolytic cleavage of a C-terminal propeptide. For the first time, a new prediction technique locating potential GPI-modification sites in precursor sequences has been applied for large-scale protein sequence database searches. The composite prediction function (with separate parametrisation for metazoan and protozoan proteins) consists of terms evaluating both amino acid type preferences at sequence positions near a supposed omega-site as well as the concordance with general physical properties encoded in multi-residue correlation within the motif sequence. The latter terms are especially successful in rejecting non-appropriate sequences from consideration. The algorithm has been validated with a self-consistency and two jack-knife tests for the learning set of fully annotated sequences from the SWISS-PROT database as well as with a newly created database "big-Pi" (more than 300 GPI-motif mutations extracted from original literature sources). The accuracy of predicting the effect of mutations in the GPI sequence motif was above 83 %. Lists of potential precursor proteins which are non-annotated in SWISS-PROT and SPTrEMBL are presented on the WWW-page http://www.embl-heidelberg.de/beisenha/gpi/gpi_p rediction. html The algorithm has been implemented in the prototype software "big-Pi predictor" which may find application as a genome annotation and target selection tool. Copyright 1999 Academic Press.
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              Multiple roles of carbonic anhydrase in cellular transport and metabolism.

              Carbonic anhydrase (CA) is a central enzyme to both transport and metabolic processes at the cellular level. In metabolically active tissue such as muscle, CA in the cytoplasm and on the sarcolemma appears to be important in facilitating CO2 transport out of the cell. Membrane-associated CA, with an extracellular orientation, also appears to be important in acidifying the outer boundary layer through the catalyzed hydration of excreted CO2. This facilitates cellular ammonia transport by providing H+ ions for the protonation of NH3, thus maintaining the trans-membrane NH3 gradient. Mitochondrial CA is known to supply HCO3- for the initial reactions of gluconeogenesis and ureagenesis in mammalian tissues, but systematic comparative studies of CA as a metabolic enzyme are lacking. CA probably evolved as an enzyme of trans-membrane facilitated CO2 transport and took on a secondary metabolic role later in metazoan evolution.
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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
                19 October 2016
                2016
                : 11
                : 10
                : e0164396
                Affiliations
                [1 ]Key Laboratory of Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China
                [2 ]Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China
                Shanghai Ocean University, CHINA
                Author notes

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

                • Conceptualization: XL JK.

                • Formal analysis: XL.

                • Investigation: XL PD.

                • Project administration: JK.

                • Resources: PD BXC KL.

                • Supervision: SL XHM.

                • Validation: XL.

                • Writing – original draft: XL.

                • Writing – review & editing: XL.

                Article
                PONE-D-16-21844
                10.1371/journal.pone.0164396
                5070816
                27760162
                41bc5d11-4249-49fd-ab66-a408d9ec372e
                © 2016 Lu 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
                : 31 May 2016
                : 23 September 2016
                Page count
                Figures: 5, Tables: 5, Pages: 24
                Funding
                This work was supported by the National Natural Science Foundation of China (31302179), the Taishan Scholar Program For Seed Industry, the Introduction of International Advanced Agricultural Science and Technology Plan (2016-X39), Shangdong Province Young and Middle-Aged Scientists Research Awards Fund (BS2015NT010), the Special Fund for Basic Scientific Research Business of Central Public Research Institutes (2015B04XK01), and the Guangdong Province Aquaculture Improved Variety Establishment of System Project.
                Categories
                Research Article
                Physical Sciences
                Chemistry
                Chemical Compounds
                Ammonia
                Biology and Life Sciences
                Immunology
                Immune Response
                Medicine and Health Sciences
                Immunology
                Immune Response
                Biology and Life Sciences
                Organisms
                Animals
                Invertebrates
                Arthropoda
                Crustaceans
                Shrimp
                Biology and Life Sciences
                Genetics
                Gene Expression
                Biology and Life Sciences
                Toxicology
                Toxicity
                Medicine and Health Sciences
                Pathology and Laboratory Medicine
                Toxicology
                Toxicity
                Biology and Life Sciences
                Biochemistry
                Metabolism
                Nitrogen Metabolism
                Biology and Life Sciences
                Genetics
                Gene Expression
                Gene Regulation
                Biology and Life Sciences
                Biochemistry
                Metabolism
                Metabolic Pathways
                Custom metadata
                All relevant data are within the paper and its Supporting Information files.

                Uncategorized
                Uncategorized

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