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De novo characterization of the liver transcriptome of javelin goby Synechogobius hasta and analysis of its transcriptomic profile following waterborne copper exposure

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Abstract

Previous studies have investigated the physiological responses to chronic copper (Cu) exposure in the liver of Synechogobius hasta; however, little information is available on the underlying molecular mechanisms. In an effort to better understand the mechanisms of Cu toxicity and to illuminate global gene expression patterns modulated by Cu exposure, we obtained the liver transcriptome information of S. hasta by RNA sequencing (RNA-seq) technology and also investigated the differential expression of genes following waterborne Cu exposure. Using the Illumina sequencing platform, as many as 60,217 unigenes were generated, with 815 bp of average length and 1298 bp of unigene N50 after filtering and assembly. For functional annotation analysis, 34,860, 31,526, 31,576, 25,808, 11,542, and 21,721 unigenes were annotated to the NR, NT, Swiss-Prot, KEGG, COG, and GO databases, respectively, and total annotation unigenes were 37,764. After 30 days of exposure to 55 μg Cu/l, a total of 292 and 1076 genes were significantly up- and down-regulated, respectively. By KEGG analysis, 660 had a specific pathway annotation. Subsequent bioinformatics analysis revealed that the differentially expressed genes were mainly related to lipid metabolism, immune system, apoptosis, and signal transduction, suggesting that these signaling pathways may be regulated by Cu exposure. The present study provides comprehensive sequence information for subsequent gene expression studies regarding S. hasta, and the transcriptome profiling after Cu exposure is also expected to improve our understanding of the molecular toxicology of Cu.

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References

  • Ali A, Rexroad CE, Thorgaard GH, Yao J, Salem M (2014) Characterization of the rainbow trout spleen transcriptome and identification of immune-related genes. Front Genet 5:348

    Article  PubMed  PubMed Central  Google Scholar 

  • Amaya MJ, Nathanson MH (2013) Calcium signaling in the liver. Compr Physiol 3:515–539

    PubMed  PubMed Central  Google Scholar 

  • Arnal N, Dominici L, De Tacconi MJ, Marra CA (2014) Copper-induced alterations in rat brain depends on route of overload and basal copper levels. Nutrition 30:96–106

    Article  CAS  PubMed  Google Scholar 

  • Audic S, Claverie JM (1997) The significance of digital gene expression profiles. Genome Res 7:986–995

    CAS  PubMed  Google Scholar 

  • Benjamini Y, Yekutieli D (2001) The control of the false discovery rate in multiple testing under dependency. Ann Stat 29:1165–1188

    Article  Google Scholar 

  • Borroni EM, Mantovani A, Locati M, Bonecchi R (2010) Chemokine receptors intracellular trafficking. Pharmacol Ther 127:1–8

    Article  CAS  PubMed  Google Scholar 

  • Bratton SB, Cohen GM (2001) Apoptotic death sensor: an organelle’s alter ego? Trends Pharmacol Sci 22:306–315

    Article  CAS  PubMed  Google Scholar 

  • Caffrey DR, O’neill LA, Shields DC (1999) The evolution of the MAP kinase pathways: coduplication of interacting proteins leads to new signaling cascades. J Mol Evol 49:567–582

    Article  CAS  PubMed  Google Scholar 

  • Chen QL, Luo Z, Zheng JL, Li XD, Liu CX, Zhao YH, Gong Y (2012) Protective effects of calcium on copper toxicity in Pelteobagrus fulvidraco: copper accumulation, enzymatic activities, histology. Ecotoxicol Environ Saf 76:126–134

    Article  CAS  PubMed  Google Scholar 

  • Chen QL, Luo Z, Liu X, Song YF, Liu CX, Zheng JL, Zhao YH (2013a) Effects of waterborne chronic copper exposure on hepatic lipid metabolism and metal-element composition in Synechogobius hasta. Arch Environ Contam Toxicol 64:301–315

    Article  CAS  PubMed  Google Scholar 

  • Chen QL, Luo Z, Pan YX, Zheng JL, Zhu QL, Sun LD, Zhuo MQ, Hu W (2013b) Differential induction of enzymes and genes involved in lipid metabolism in liver and visceral adipose tissue of juvenile yellow catfish Pelteobagrus fulvidraco exposed to copper. Aquat Toxicol 136:72–78

    PubMed  Google Scholar 

  • Chen QL, Luo Z, Wu K, Huang C, Zhuo MQ, Song YF, Hu W (2015) Differential effects of dietary copper deficiency and excess on lipid metabolism in yellow catfish Pelteobagrus fulvidraco. Comp Biochem Physiol 184B:19–28

    Article  Google Scholar 

  • Collins T, Read M, Neish A, Whitley M, Thanos D, Maniatis T (1995) Transcriptional regulation of endothelial cell adhesion molecules: NF-kappa B and cytokine-inducible enhancers. FASEB J 9:899–909

    CAS  PubMed  Google Scholar 

  • Collins SA, Sinclair G, Mcintosh S, Bamforth F, Thompson R, Sobol I, Osborne G, Corriveau A, Santos M, Hanley B (2010) Carnitine palmitoyltransferase 1A (CPT1A) P479L prevalence in live newborns in Yukon, Northwest Territories, and Nunavut. Mol Genet Metab 101:200–204

    Article  CAS  PubMed  Google Scholar 

  • Conesa A, Gotz S, Garcia-Gomez JM, Terol J, Talon M, Robles M (2005) Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research. Bioinformatics 21:3674–3676

    Article  CAS  PubMed  Google Scholar 

  • Dethloff GM, Schlenk D, Hamm JT, Bailey HC (1999) Alterations in physiological parameters of rainbow trout (Oncorhynchus mykiss) with exposure to copper and copper/zinc mixtures. Ecotoxicol Environ Saf 42:253–264

    Article  CAS  PubMed  Google Scholar 

  • Ebner HL, Blatzer M, Nawaz M, Krumschnabel G (2007) Activation and nuclear translocation of ERK in response to ligand-dependent and-independent stimuli in liver and gill cells from rainbow trout. J Exp Biol 210:1036–1045

    Article  CAS  PubMed  Google Scholar 

  • Ekblom R, Galindo J (2011) Applications of next generation sequencing in molecular ecology of non-model organisms. Heredity 107:1–15

    Article  CAS  PubMed  Google Scholar 

  • Ellis RE, Yuan J, Horvitz H (1991) Mechanisms and functions of cell death. Annu Rev Cell Biol 7:663–698

    Article  CAS  PubMed  Google Scholar 

  • Frawley R, White K, Brown R, Musgrove D, Walker N, Germolec D (2011) Gene expression alterations in immune system pathways in the thymus after exposure to immunosuppressive chemicals. Environ Health Perspect 119:371–376

    Article  CAS  PubMed  Google Scholar 

  • Fromenty B, Pessayre D (1995) Inhibition of mitochondrial beta-oxidation as a mechanism of hepatotoxicity. Pharmacol Ther 67:101–154

    Article  CAS  PubMed  Google Scholar 

  • Giacomin M, Jorge MB, Bianchini A (2014) Effects of copper exposure on the energy metabolism in juveniles of the marine clam Mesodesma mactroides. Aquat Toxicol 152:30–37

    Article  CAS  PubMed  Google Scholar 

  • Gotz S, Garcia-Gomez JM, Terol J, Williams TD, Nagaraj SH, Nueda MJ, Robles M, Talon M, Dopazo J, Conesa A (2008) High-throughput functional annotation and data mining with the Blast2GO suite. Nucleic Acids Res 36:3420–3435

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng Q (2011) Full-length transcriptome assembly from RNA-seq data without a reference genome. Nat Biotechnol 29:644–652

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gui D, Jia K, Xia J, Yang L, Chen J, Wu Y, Yi M (2013) De novo assembly of the indo-pacific humpback dolphin leucocyte transcriptome to identify putative genes involved in the aquatic adaptation and immune response. PLoS One 8:e72417

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guo H, Ye CX, Wang AL, Xian JA, Liao SA, Miao YT, Zhang SP (2013) Transcriptome analysis of the Pacific white shrimp Litopenaeus vannamei exposed to nitrite by RNA-seq. Fish Shellfish Immunol 35:2008–2016

    Article  CAS  PubMed  Google Scholar 

  • Harding LB, Schultz IR, Goetz GW, Luckenbach JA, Young G, Goetz FW, Swanson P (2013) High-throughput sequencing and pathway analysis reveal alteration of the pituitary transcriptome by 17α-ethynylestradiol (EE2) in female coho salmon, Oncorhynchus kisutch. Aquat Toxicol 142:146–163

    Article  PubMed  Google Scholar 

  • He S, Liang XF, Sun J, Li L, Yu Y, Huang W, Qu CM, Cao L, Bai XL, Tao YX (2013) Insights into food preference in hybrid F1 of Siniperca chuatsi (♀)× Siniperca scherzeri (♂) mandarin fish through transcriptome analysis. BMC Genom 14:601

    Article  CAS  Google Scholar 

  • Hovhannisyan LP, Mkrtchyan GM, Sukiasian SH, Boyajyan AS (2010) Alterations in the complement cascade in post-traumatic stress disorder. Allergy Asthma Clin Immunol 6:3

    Article  PubMed  PubMed Central  Google Scholar 

  • Huang Q, Dong S, Fang C, Wu X, Ye T, Lin Y (2012) Deep sequencing-based transcriptome profiling analysis of Oryzias melastigma exposed to PFOS. Aquat Toxicol 120:54–58

    Article  PubMed  Google Scholar 

  • Huang C, Chen QL, Luo Z, Shi X, Pan YX, Song YF, Zhuo MQ, Wu K (2014) Time-dependent effects of waterborne copper exposure influencing hepatic lipid deposition and metabolism in javelin goby Synechogobius hasta and their mechanism. Aquat Toxicol 155:291–300

    Article  CAS  PubMed  Google Scholar 

  • Kanehisa M, Goto S (2000) KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res 28:27–30

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kim EK, Choi EJ (2010) Pathological roles of MAPK signaling pathways in human diseases. Biochim Biophys Acta 1802:396–405

    Article  CAS  PubMed  Google Scholar 

  • Kobayashi Y, Miyazawa M, Kamei A, Abe K, Kojima T (2010) Ameliorative effects of mulberry (Morus alba L.) leaves on hyperlipidemia in rats fed a high-fat diet: induction of fatty acid oxidation, inhibition of lipogenesis, and suppression of oxidative stress. Biosci Biotechnol Biochem 74:2385–2395

    Article  CAS  PubMed  Google Scholar 

  • Lewis J, Wataha J, Mccloud V, Lockwood P, Messer R, Tseng WY (2005) Au (III), Pd (II), Ni (II), and Hg(II) alter NFκB signaling in THP1 monocytic cells. J Biomed Mater Res A 74:474–481

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Luo Z, Xiong B, Liu X, Zhao Y, Hu G, Lv G (2010) Effect of waterborne copper exposure on growth, hepatic enzymatic activities and histology in Synechogobius hasta. Ecotoxicol Environ Saf 73:1286–1291

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−∆∆CT method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  • Long Y, Li Q, Zhou B, Song G, Li T, Cui Z (2013) De novo assembly of mud loach (Misgurnus anguillicaudatus) skin transcriptome to identify putative genes involved in immunity and epidermal mucus secretion. PLoS One 8:e56998

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Marioni JC, Mason CE, Mane SM, Stephens M, Gilad Y (2008) RNA-seq: an assessment of technical reproducibility and comparison with gene expression arrays. Genome Res 18:1509–1517

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mattie MD, McElwee MK, Freedman JH (2008) Mechanism of copper-activated transcription: activation of AP-1, and the JNK/SAPK and p38 signal transduction pathways. J Mol Biol 383:1008–1018

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McElwee MK, Song MO, Freedman JH (2009) Copper activation of NF-κB signaling in HepG2 cells. J Mol Biol 393:1013–1021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mortazavi A, Williams BA, Mccue K, Schaeffer L, Wold B (2008) Mapping and quantifying mammalian transcriptomes by RNA-seq. Nat Methods 5:621–628

    Article  CAS  PubMed  Google Scholar 

  • Mu Y, Li M, Ding F, Ding Y, Ao J, Hu S, Chen X (2014) De novo characterization of the spleen transcriptome of the large yellow croaker (Pseudosciaena crocea) and analysis of the immune relevant genes and pathways involved in the antiviral response. PLoS One 9:e97471

    Article  PubMed  PubMed Central  Google Scholar 

  • Nawaz M, Manzl C, Lacher V, Krumschnabel G (2006) Copper-induced stimulation of extracellular signal-regulated kinase in trout hepatocytes: the role of reactive oxygen species, Ca2+, and cell energetics and the impact of extracellular signal-regulated kinase signaling on apoptosis and necrosis. Toxicol Sci 92:464–475

    Article  CAS  PubMed  Google Scholar 

  • Nielsen HD, Brown MT, Brownlee C (2003) Cellular responses of developing Fucus serratus embryos exposed to elevated concentrations of Cu2+. Plant, Cell Environ 26:1737–1747

    Article  CAS  Google Scholar 

  • Pierron F, Normandeau E, Defo MA, Campbell PG, Bernatchez L, Couture P (2011) Effects of chronic metal exposure on wild fish populations revealed by high-throughput cDNA sequencing. Ecotoxicology 20:1388–1399

    Article  CAS  PubMed  Google Scholar 

  • Pietrobon D, Virgilio F, Pozzan T (1990) Structural and functional aspects of calcium homeostasis in eukaryotic cells. Eur J Biochem 193:599–622

    Article  CAS  PubMed  Google Scholar 

  • Ramesh GT, Manna SK, Aggarwal BB, Jadhav AL (1999) Lead activates nuclear transcription factor-κB, activator protein-1, and amino-terminal c-Jun kinase in pheochromocytoma cells. Toxicol Appl Pharmacol 155:280–286

    Article  CAS  PubMed  Google Scholar 

  • Rami A, Agarwal R, Botez G, Winckler J (2000) μ-Calpain activation, DNA fragmentation, and synergistic effects of caspase and calpain inhibitors in protecting hippocampal neurons from ischemic damage. Brain Res 866:299–312

    Article  CAS  PubMed  Google Scholar 

  • Robinson CB, Wills PS, Riche MA, Straus DL (2013) Tissue-specific copper concentrations in red drum after long-term exposure to sublethal levels of waterborne copper and a 21-day withdrawal. N Am J Aquac 75:1–6

    Article  Google Scholar 

  • Santos EM, Ball JS, Williams TD, Wu H, Ortega F, Van Aerle R, Katsiadaki I, Falciani F, Viant MR, Chipman JK (2010) Identifying health impacts of exposure to copper using transcriptomics and metabolomics in a fish model. Environ Sci Technol 44:820–826

    Article  CAS  PubMed  Google Scholar 

  • Schafer S, Bickmeyer U, Koehler A (2009) Measuring Ca2+-signalling at fertilization in the sea urchin Psammechinus miliaris: alterations of this Ca2+-signal by copper and 2, 4, 6-tribromophenol. Comp Biochem Physiol C 150:261–269

    Google Scholar 

  • Seki E, Schwabe RF (2015) Hepatic inflammation and fibrosis: functional links and key pathways. Hepatology 61:1066–1079

    Article  PubMed  PubMed Central  Google Scholar 

  • Sheridan MA (1988) Lipid dynamics in fish: aspects of absorption, transportation, deposition and mobilization. Comp Biochem Physiol B 90:679–690

    CAS  PubMed  Google Scholar 

  • Song YF, Luo Z, Zhang LH, Hogstrand C, Pan YX (2016) Endoplasmic reticulum stress and disturbed calcium homeostasis are involved in copper-induced alteration in hepatic lipid metabolism in yellow catfish Pelteobagrus fulvidraco. Chemosphere 144:2443–2453

    Article  CAS  PubMed  Google Scholar 

  • Stohs S, Bagchi D (1995) Oxidative mechanisms in the toxicity of metal ions. Free Radic Biol Med 18:321–336

    Article  CAS  PubMed  Google Scholar 

  • Sun M, Zhao Y, Xu C (2008) Cross-talk between calpain and caspase-3 in penumbra and core during focal cerebral ischemia-reperfusion. Cell Mol Neurobiol 28:71–85

    Article  CAS  PubMed  Google Scholar 

  • Tassabehji NM, Vanlandingham JW, Levenson CW (2005) Copper alters the conformation and transcriptional activity of the tumor suppressor protein p53 in human Hep G2 cells. Exp Biol Med 230:699–708

    CAS  Google Scholar 

  • Tocher DR (2003) Metabolism and functions of lipids and fatty acids in teleost fish. Rev Fish Sci 11:107–184

    Article  CAS  Google Scholar 

  • Wang Z, Gerstein M, Snyder M (2009) RNA-seq: a revolutionary tool for transcriptomics. Nat Rev Genet 10:57–63

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ye J, Fang L, Zheng H, Zhang Y, Chen J, Zhang Z, Wang J, Li S, Li R, Bolund L (2006) WEGO: a web tool for plotting GO annotations. Nucleic Acids Res 34:W293–W297

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhan YJ (2005) The status quo of heavy metal pollution in Bohai sea and the abecedarian estimate of the effect of heavy metals on the growth of typical phytoplankton. Doctoral dissertation. Ocean University of China, Qingdao, China

  • Zhang R, Zhang LL, Ye X, Tian YY, Sun CF, Lu MX, Bai JJ (2013) Transcriptome profiling and digital gene expression analysis of Nile tilapia (Oreochromis niloticus) infected by Streptococcus agalactiae. Mol Biol Rep 40:5657–5668

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grants Nos. 31372547, 31422056, 31072226, and 30800850) and the Fundamental Research Funds for the Central Universities (Grant Nos. 2662015PY017, 2013PY073). We thank Beijing Genomics Institute (Shenzhen, China, http://english.big.cas.cn/) for cDNA library construction and Illumina sequencing.

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Correspondence to Zhi Luo.

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Chen, QL., Luo, Z., Huang, C. et al. De novo characterization of the liver transcriptome of javelin goby Synechogobius hasta and analysis of its transcriptomic profile following waterborne copper exposure. Fish Physiol Biochem 42, 979–994 (2016). https://doi.org/10.1007/s10695-015-0190-2

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