An integrated proteomic and metabolomic study on the gender-specific responses of mussels Mytilus galloprovincialis to tetrabromobisphenol A (TBBPA)

Tetrabromobisphenol A (TBBPA), accounting for the largest production of brominated flame-retardants (BFRs) along the Laizhou Bay in China, is of great concern due to its diverse toxicities. In this study, we focused on the gender-specific responses of TBBPA in mussel Mytilus galloprovincialis using an integrated proteomic and metabolomic approach. After exposure of TBBPA (10 mu g L-1) for one month, a total of 9 metabolites and 67 proteins were altered in mussel gills from exposed group. The significant changes of metabolites in female mussel gills from exposed group exhibited the disturbances in energy metabolism and osmotic regulation, while in male samples only be found the variation of metabolites related to osmotic regulation. iTRAQ-based proteomic analysis showed biological differences between male and female mussel gills from solvent control group. The higher levels of proteins related to primary and energy metabolism and defense mechanisms in male mussel gills meant a greater anti-stress capability of male mussels. Further analysis revealed that TBBPA exposure affected multiple biological processes consisting of production and development, material and energy metabolism, signal transduction, gene expression, defense mechanisms and apoptosis in both male and female mussels with different mechanisms. Specially, the responsive proteins of TBBPA in male mussels signified higher tolerance limits than those in female individuals, which was consistent with the biological differences between male and female mussel gills from solvent control group. This work suggested that the gender differences should be considered in ecotoxicology. (C) 2015 Elsevier Ltd. All rights reserved

A metabolomic study on the biological effects of metal pollutions in oysters Crassostrea sikamea

Metal pollution has become a great threat to organisms in the estuaries in South China. In the present study, the oysters Crassostrea sikamea were collected from one clean (Jiuzhen) and five metal polluted sites (Baijiao, Fugong, Gongqian, Jinshan and Songyu). The tissue metal concentrations in oysters indicated that the five metal sites were polluted by several metals, including Cr, Ni, Co, Cu, Zn, Ag, Cd and Pb with different patterns. Especially, Cu and Zn were the major contaminants in Baijiao, Fugong and Jinshan sites. The metabolic responses in oysters C. sikamea indicated that the metal pollutions in BJ, FG, JS and SY sites induced disturbances in osmotic regulation and energy metabolism via different metabolic pathways. However, the metal pollution in GQsite mainly influenced the osmotic regulation in the oysters C sikamea. This study demonstrates that NMR-based metabolomics is useful to characterize metabolic responses induced by metal pollution. (C) 2015 Elsevier Ltd. All rights reserved

Proteomic responses reveal the differential effects induced by cadmium in mussels Mytilus galloprovincialis at early life stages

Cadmium (Cd) has become an important metal contaminant and posed severe risk on the organisms in the coastal environments of the Bohai Sea. Marine mussel Mytilus galloprovincialis is widely distributed along the Bohai coast and consumed as seafood by local residents. Evidences indicate that the early stages of marine organisms are more sensitive to metal contaminants. In this study, we applied two-dimensional electrophoresis-based proteomics to characterize the biological effects of Cd (50 mu g L-1) in the early life stages (D-shape larval and juvenile) of mussels. The different proteomic responses demonstrated the differential responsive mechanisms to Cd exposure in these two early life stages of mussels. In details, results indicated that Cd mainly induced immune and oxidative stresses in both D-shape larval and juvenile mussels via different pathways. In addition, the significant up-regulation of triosephosphate isomerase and metallothionein confirmed the enhanced energy demand and mobilized detoxification mechanism in D-shape larval mussels exposed to Cd. In juvenile mussels, Cd exposure also induced clear apoptosis. Overall, this work suggests that Cd is a potential immune toxicant to mussel M. galloprovincialis at early life stages. (C) 2016 Elsevier Ltd. All rights reserved

Multiple biomarkers of biological effects induced by cadmium in clam Ruditapes philippinarum

Cadmium (Cd) is a known heavy metal pollutant in the Bohai Sea. Manila clam Ruditapes philippinarum is an important fishery species along the Bohai coast. In this study, the biological effects induced by two concentrations (20 and 200 mu g/L) of Cd were characterized using multiple biochemical indices in the digestive glands of clam R. philippinarum. The total hemocyte counts, reactive oxygen species productions and antioxidant enzyme activities exhibited that Cd induced dose-dependent immune and oxidative stresses in clam digestive glands. Metabolic responses indicated that both Cd exposures caused immune stress marked by the elevated branched chain amino acids (valine, leucine and isoleucine), together with the disturbance in energy metabolism. The differential metabolic biomarkers related to osmotic stress, including homarine, betaine, tyrosine and phenylalanine, suggested the differential responsive mechanisms in clam digestive glands induced by Cd exposures. In addition, both Cd treatments enhanced the anaerobiosis metabolism in clam digestive glands via differential metabolic pathways. (c) 2015 Elsevier Ltd. All rights reserved.Cadmium (Cd) is a known heavy metal pollutant in the Bohai Sea. Manila clam Ruditapes philippinarum is an important fishery species along the Bohai coast. In this study, the biological effects induced by two concentrations (20 and 200 mu g/L) of Cd were characterized using multiple biochemical indices in the digestive glands of clam R. philippinarum. The total hemocyte counts, reactive oxygen species productions and antioxidant enzyme activities exhibited that Cd induced dose-dependent immune and oxidative stresses in clam digestive glands. Metabolic responses indicated that both Cd exposures caused immune stress marked by the elevated branched chain amino acids (valine, leucine and isoleucine), together with the disturbance in energy metabolism. The differential metabolic biomarkers related to osmotic stress, including homarine, betaine, tyrosine and phenylalanine, suggested the differential responsive mechanisms in clam digestive glands induced by Cd exposures. In addition, both Cd treatments enhanced the anaerobiosis metabolism in clam digestive glands via differential metabolic pathways. (c) 2015 Elsevier Ltd. All rights reserved

Proteomic and metabolomic analysis on the toxicological effects of As (III) and As (V) in juvenile mussel Mytilus galloprovincialis

Inorganic arsenic (As) is a known pollutant including two chemical forms (arsenite (As III) and arsenate (As V)), in marine and coastal environment. Marine mussel Mytilus galloprovincialis is an important environmental monitoring species around the world. In this study, we focused on valence-specific responses of As in juvenile mussel M. galloprovincialis using a combined proteomic and metabolomic approach. Metabolic responses indicated that As (III) mainly caused disturbance in osmotic regulation in juvenile mussels. As (V) caused disturbances in both osmotic regulation and energy metabolism marked by different metabolic responses, including betaine, taurine, glucose and glycogen. Proteomic responses exhibited that As (III) had a significant negative effect on cytoskeleton and cell structure (actin and collagen alpha-6(VI) chain). As (V) affected some key enzymes involved in energy metabolism (cytosolic malate dehydrogenase, cMDH) and cell development (ornithine aminotransferase and astacin). Overall, all these results confirmed the valence-specific responses in juvenile mussels to As exposures. These findings demonstrate that a combined metabolomic and proteomic approach could provide an important insight into the toxicological effects of environmental pollutants in organisms. (C) 2016 Elsevier Ltd. All rights reserved

Homologous cloning, characterization and expression of a new halophyte phytochelatin synthase gene in Suaeda salsa

The halophyte Suaeda salsa can grow in heavy metal-polluted areas along intertidal zones having high salinity. Since phytochelatins can effectively chelate heavy metals, it was hypothesized that S. salsa possessed a phytochelatin synthase (PCS) gene. In the present study, the cDNA of PCS was obtained from S. salsa (designated as SsPCS) using homologous cloning and the rapid amplification of cDNA ends (RACE). A sequence analysis revealed that SsPCS consisted of 1 916 bp nucleotides, encoding a polypeptide of 492 amino acids with one phytochelatin domain and one phytochelatin C domain. A similarity analysis suggested that SsPCS shared up to a 58.6% identity with other PCS proteins and clustered with PCS proteins from eudicots. There was a new kind of metal ion sensor motif in its C-terminal domain. The SsPCS transcript was more highly expressed in elongated and fibered roots and stems (P < 0.05) than in leaves. Lead and mercury exposure significantly enhanced the mRNA expression of SsPCS (P < 0.05). To the best of our knowledge, SsPCS is the second PCS gene cloned from a halophyte, and it might contain a different metal sensing capability than the first PCS from Thellungiella halophila. This study provided a new view of halophyte PCS genes in heavy metal tolerance

A comparative proteomic study on the effects of metal pollution in oysters Crassostrea hongkongensis

The metal pollution has posed great risk on the coastal organisms along the Jiulongjiang Estuary in South China. In this work, two-dimensional electrophoresis-based proteomics was applied to the oysters Crassostrea hongkongensis from metal pollution sites to characterize the proteomic responses to metal pollution. Metal accumulation and proteomic responses indicated that the oysters from BJ site were more severely contaminated than those from FG site. Compared with those oyster samples from the clean site (JZ), metal pollution induced cellular injuries, oxidative and immune stresses in oyster heapatopancreas from both BJ and FG sites via differential metabolic pathways. In addition, metal pollution in BJ site induced disturbance in energy and lipid metabolisms in oysters. Results indicated that cathepsin L and ferritin GF1 might be the biomarkers of As and Fe in oyster C. hongkongensis, respectively. This study demonstrates that proteomics is a useful tool for investigating biological effects induced by metal pollution. (C) 2016 Elsevier Ltd. All rights reserved

iTRAQ-based quantitative proteomic analyses on the gender-specific responses in mussel Mytilus galloprovincialis to tetrabromobisphenol A

Tetrabromobisphenol A (TBBPA) accounts for the largest production of brominated flame-retardants (BFRs) along the Laizhou Bay in China and is the most widely used BFR in industrial products. It can induce diverse toxicities including hepatotoxicity, nephrotoxicity, neurotoxicity and endocrine disrupting effects in mammalian and fish models. In this work, we applied iTRAQ-based proteomics to investigate the gender-specific responses in mussel Mytilus galloprovincialis to TBBPA. Thirty-one proteins were differentially expressed in hepatopancreas between male and female mussels, which clearly indicated the biological differences between male and female mussels at the protein level. After exposure of TBBPA (18.4 nmol/L) for one month, a total of 60 proteins were differentially expressed in response to the TBBPA treatment in mussel hepatopancreas, among which 33 and 29 proteins were significantly altered in TBBPA-treated male and female mussel samples, respectively. Only two of the 60 proteins were commonly altered in both male and female mussel samples exposed to TBBPA. Based on KEGG analysis, these differentially expressed proteins of TBBPA-induced effects were assigned to several groups, including cytoskeleton, reproduction and development, metabolism, signal transduction, gene expression, stress response and apoptosis. Overall, results indicated that TBBPA exposure could induce apoptosis, oxidative and immune stresses and disruption in energy, protein and lipid metabolisms in both male and female mussels with different mechanisms. This work suggested that the gender differences should be considered in ecotoxicoproteomics. (C) 2014 Elsevier B.V. All rights reserved.Tetrabromobisphenol A (TBBPA) accounts for the largest production of brominated flame-retardants (BFRs) along the Laizhou Bay in China and is the most widely used BFR in industrial products. It can induce diverse toxicities including hepatotoxicity, nephrotoxicity, neurotoxicity and endocrine disrupting effects in mammalian and fish models. In this work, we applied iTRAQ-based proteomics to investigate the gender-specific responses in mussel Mytilus galloprovincialis to TBBPA. Thirty-one proteins were differentially expressed in hepatopancreas between male and female mussels, which clearly indicated the biological differences between male and female mussels at the protein level. After exposure of TBBPA (18.4 nmol/L) for one month, a total of 60 proteins were differentially expressed in response to the TBBPA treatment in mussel hepatopancreas, among which 33 and 29 proteins were significantly altered in TBBPA-treated male and female mussel samples, respectively. Only two of the 60 proteins were commonly altered in both male and female mussel samples exposed to TBBPA. Based on KEGG analysis, these differentially expressed proteins of TBBPA-induced effects were assigned to several groups, including cytoskeleton, reproduction and development, metabolism, signal transduction, gene expression, stress response and apoptosis. Overall, results indicated that TBBPA exposure could induce apoptosis, oxidative and immune stresses and disruption in energy, protein and lipid metabolisms in both male and female mussels with different mechanisms. This work suggested that the gender differences should be considered in ecotoxicoproteomics. (C) 2014 Elsevier B.V. All rights reserved

Differential metabolic responses in three life stages of mussels Mytilus galloprovincialis exposed to cadmium

Cadmium (Cd) is one of the most important metal contaminants in the Bohai Sea. In this work, NMR-based metabolomics was used to investigate the toxicological effects of Cd at an environmentally relevant concentration (50 A mu g L-1) in three different life stages (D-shape larval, juvenile and adult) of mussels Mytilus galloprovincialis. Results indicated that the D-shape larval mussel was the most sensitive life stage to Cd. The significantly different metabolic profiles meant that Cd induced differential toxicological effects in three life stages of mussels. Basically, Cd caused osmotic stress in all the three life stages via different metabolic pathways. Cd exposure reduced the anaerobiosis in D-shape larval mussels and disturbed lipid metabolism in juvenile mussels, respectively. Compared with the D-shape larval and juvenile mussels, the adult mussels reduced energy consumption to deal with Cd stress

Metabolomic analysis revealed that female mussel Mytilus galloprovincialis was sensitive to bisphenol A exposures

Bisphenol A (BPA) is a synthetic compound used in numerous chemicals, such as polycarbonate plastics and epoxy resins, and it can be released into aquatic environment and poses risk on aquatic organisms. In this work, metabolomics was applied to characterize the metabolic responses in mussel Mytilus galloprovincialis exposed to BPA. Our results indicated that the gonad of female mussel was sensitive to BPA exposures (1 and 10 mu g/L) for one month. However, no significant metabolic responses were observed in male mussel gonads exposed to these two concentrations of BPA. Overall, this limited study suggested that the gender differences should be considered in marine ecotoxicology. (C) 2014 Elsevier B.V. All rights reserved.Bisphenol A (BPA) is a synthetic compound used in numerous chemicals, such as polycarbonate plastics and epoxy resins, and it can be released into aquatic environment and poses risk on aquatic organisms. In this work, metabolomics was applied to characterize the metabolic responses in mussel Mytilus galloprovincialis exposed to BPA. Our results indicated that the gonad of female mussel was sensitive to BPA exposures (1 and 10 mu g/L) for one month. However, no significant metabolic responses were observed in male mussel gonads exposed to these two concentrations of BPA. Overall, this limited study suggested that the gender differences should be considered in marine ecotoxicology. (C) 2014 Elsevier B.V. All rights reserved