Glutathione peroxidase from the white shrimp Litopenaeus vannamei: Characterization and its regulation upon pH and Cd exposure
Key Laboratory of Ecology and Environmental Science in Guangdong Higher Education, College of Life Science, South China Normal University, Guangzhou, People's Republic of China. Ecotoxicology
(Impact Factor: 2.71).
06/2012; 21(6):1585-92. DOI: 10.1007/s10646-012-0942-z
Glutathione peroxidase (GPx) is a key enzyme of cellular detoxification systems that defend cells against reactive oxygen species. Knowledge of the complement of GPx in shrimp is essential to understanding regulation and detoxification mechanisms of environmental stress. In this study, we expressed GPx from white shrimp Litopenaeus vannamei in Escherichia coli, and then characterized the purified recombinant enzyme with respect to the effects of pH, temperature on its catalytic activity. Quantitative real-time PCR and western blot analysis were carried out to investigate the expression patterns of GPx in shrimp hepatopancreas exposed to Cd stress. A statistically significant increase in expression of GPx mRNA and protein was observed in the Cd stress at 24 h. By contrast, western blot showed a significant up-regulation in GPx protein expression at 12 h exposed pH stress (5.6 and 9.3, respectively). These results suggest that L. vannamei GPx expression was modulated by Cd and pH stress and may play an important role in detoxification of xenobiotics and antioxidant defense. We conclude that GPx could be used as biomarkers of Cd and pH stress in aquatic environment for the studied species.
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ABSTRACT: The purpose of this study was to investigate the toxic effects induced by benzo[a]pyrene (BaP). In the present study, juvenile white shrimp Litopenaeus vannamei were exposed to BaP for 21 days at four different concentrations as 0, 0.03, 0.3 and 3 μg/L. Detoxification system parameters, transcription of metabolism-related genes, and damage indexes were investigated for screening. It showed mRNA expression levels of superoxide dismutase (SOD), cytochrome P450 (CYP) 1A1 and glutathione-S-transferase (GST) in hepatopancreas were significantly induced at day 3 by 0.3 and 3 μg/L BaP, and returned to the initial level afterwards. 7-Ethoxyresorufin O-deethylase (EROD), GST and SOD activities in gill and hepatopancreas were also induced by 0.3 and 3 μg/L BaP, while T-AOC and GSH contents were reduced after BaP exposure. The study also showed that 0.3 and 3 μg/L BaP caused damage effects, while 0.03 μg/L BaP treatment did not exhibit any damage effects. Moreover, according to the correlation analysis results, we conclude that EROD activity, GST activity and GSH content in hepatopancreas can be used as defense biomarkers; DNA strand breaks, lipid peroxidation (LPO) level and protein carbonyl (PC) content in gill and hepatopancreas can be used as damage biomarkers; EROD activity, GST activity, GSH contents and damage effects of DNA strand breaks, LPO level and PC content in hepatopancreas can be used as combined biomarkers. These results will provide information not only on potential biomarkers that could be effectively applied to biomonitor aquatic environment to contamination, but also provide information on toxic effects of this specie.
Environmental Toxicology and Pharmacology 08/2014; 39(1). DOI:10.1016/j.etap.2014.08.006 · 2.08 Impact Factor
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ABSTRACT: Cadmium, one of the most toxic heavy metals in aquatic environments, has severe effects on marine invertebrates and fishes. The MAPK signaling pathway plays a vital role in stress responses of animals. The mitogen-activated protein kinase (MAPK) signaling pathway plays a vital role in animals' stress responses, including mediation of apoptosis induced by the Rho GTPase Cdc42. However, there is limited knowledge about its function in shrimps, although disorders exacerbated by environmental stresses (including heavy metal pollution) have caused serious mortality in commercially cultured shrimps. Thus, we probed roles of Cdc42 in Litopenaeus vannamei shrimps (LvCdc42) during cadmium exposure by inhibiting its expression using dsRNA-mediated RNA interference. The treatment successfully reduced expression levels of MAPKs (including p38, JNK, and ERK). Cadmium exposure induced significant increases in expression levels of LvCdc42 and MAPKs, accompanied by reductions in total hemocyte counts (THC) and increases in apoptotic hemocyte ratios and ROS production. However, all of these responses were much weaker in LvCdc42-suppressed shrimps, in which mortality rates were higher than in controls. Our results suggest that the MAPK pathway plays a vital role in shrimps' responses to Cd(2+). They also indicate that LvCdc42 in shrimps participates in its regulation, and thus plays key roles in ROS production, regulation of apoptosis and associated stress responses.
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Aquatic toxicology (Amsterdam, Netherlands) 04/2015; 163. DOI:10.1016/j.aquatox.2015.03.023 · 3.45 Impact Factor
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ABSTRACT: The growth, activation of immunity, immune parameters, and transcript levels of cytMnSOD, mtMnSOD, ecCuZnSOD, glutathione peroxidase (GPx), catalase, lysozyme, and penaeidin 3a were examined in white shrimp Litopenaeus vannamei reared at pH 6.8 and 8.1 after 24 weeks. No significant difference in growth was observed between the two groups. An in vitro study indicated that phenoloxidase activity and respiratory bursts (RB, release of the superoxide anion) were significantly higher in the haemocytes of pH 8.1 shrimp (shrimp reared at pH 8.1) than in pH 6.8 shrimp (shrimp reared at pH 6.8). An in vivo study indicated that the levels of immune parameters of pH 8.1 shrimp were significantly higher than in pH 6.8 shrimp, and the transcript levels of cytMnSOD, ecCuZnSOD, glutathione peroxidase, lysozyme, and penaeidin 3a were down-regulated in pH 6.8 shrimp. In another experiment, shrimp reared at pH 6.8 and 8.1 for 24 weeks were challenged with Vibrio alginolyticus. The mortality rate of pH 6.8 shrimp was significantly higher than in pH 8.1 shrimp over 12∼168 h. Phagocytic activity, phagocytic index, and clearance efficiency to V. alginolyticus were significantly lower in pH 6.8 shrimp. We concluded that shrimp under long-term culture at pH 6.8 exhibited decreased resistance against V. alginolyticus as evidenced by reductions in the activation of immunity and immune parameters together with decreased transcript levels of cytMnSOD, ecCuZnSOD, GPx, lysozyme, and penaeidin 3a.
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Fish & Shellfish Immunology 06/2015; 46(2). DOI:10.1016/j.fsi.2015.05.055 · 2.67 Impact Factor
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