Biochemical responses in mussels Perna perna exposed to diesel B5

Nogueira, Lílian; Garcia, Danielly; Trevisan, Rafael; Sanches, Ana Letícia Madeira; Acosta, Daiane da Silva; Dafre, Alcir Luiz; Oliveira, Thiago Y.; Almeida, Eduardo Alves de

doi:10.1016/j.chemosphere.2015.04.034

Cited by 17 publications

(9 citation statements)

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“…Interestingly, MDA levels decreased after exposure (24 h) to the highest PHE concentration, remaining low even after DEP. This pattern has already been reported with other aquatic species exposed to PAH mixtures (Arantes Felício et al, 2015;Nogueira et al, 2015) and on a time-dependent oxidative damage (Ansaldo et al, 2005;Lüchmann et al, 2011Lüchmann et al, , 2014. At the same time, the absence of increased MDA levels along with PHE bioaccumulation could indicate a low acute toxicity of this compound to N. nodosus.…”

Section: Discussionsupporting

confidence: 83%

Exposure to phenanthrene and depuration: Changes on gene transcription, enzymatic activity and lipid peroxidation in gill of scallops Nodipecten nodosus

et al. 2016

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Understanding the mechanism of phenanthrene (PHE) biotransformation and related cellular responses in bivalves can be an important tool to elucidate the risks of polycyclic aromatic hydrocarbons (PAHs) to aquatic organisms. In the present study it was analyzed the transcriptional levels of 13 biotransformation genes related to cytochrome P450 (CYP), glutathione S-transferase (GST), sulfotransferase (SULT), flavin-containing monooxygenase and fatty acid-binding proteins by qPCR in gill of scallops Nodipecten nodosus exposed for 24 or 96h to 50 or 200μgL(-1) PHE (equivalent to 0.28 and 1.12μM, respectively), followed by depuration in clean water for 96h (DEP). Likewise, it was quantified the activity of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), glutathione reductase (GR), glucose 6-phosphate dehydrogenase (G6PDH), GST and levels of lipid peroxidation. Increased transcriptional levels of CYP2UI-like, CYP2D20-like, CYP3A11-like, GSTomega-like, SULT1B1-like genes were detected in organisms exposed to PHE for 24 or 96h. In parallel, GR and GPX activities increased after 96h exposure to 200μgL(-1) PHE and G6PDH activity increased after 24h exposure to 50μgL(-1) PHE. This enhancement of antioxidant and phase I and II biotransformation systems may be related to the 2.7 and 12.5 fold increases in PHE bioaccumulation after 96h exposure to 50 and 200μgL(-1) PHE, respectively. Interestingly, DEP caused reestablishment of GPX and GR activity, as well as to the transcript levels of all upregulated biotransformation genes (except for SULT1B1-like). Bioaccumulated PHE levels decreased 2.5-2.9 fold after depuration, although some biochemical and molecular modifications were still present. Lipid peroxidation levels remained lower in animals exposed to 200μgL(-1) PHE for 24h and DEP. These data indicate that N. nodosus is able to induce an antioxidant and biotransformation-related response to PHE exposure, counteracting its toxicity, and DEP can be an effective protocol for bivalve depuration after PHE exposure.

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Section: Discussionsupporting

confidence: 83%

Exposure to phenanthrene and depuration: Changes on gene transcription, enzymatic activity and lipid peroxidation in gill of scallops Nodipecten nodosus

et al. 2016

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“…These characteristics, in addition to their widespread distribution and significant economic importance, have made bivalves an ideal model organism in aquatic toxicology and ecotoxicology studies to investigate and characterize the effects and mechanisms of toxicity for several pollutants (Apeti et al, 2010;Depledge and Fossi, 1994). For instance, studies have investigated the modulation of major defense systems (e.g., antioxidant and biotransformation) in bivalves exposed to environmental pollutants (Aloıísio Torres et al, 2002;Borković et al, 2005;Dafre et al, 2004;de Almeida et al, 2004;Doyotte et al, 1997;Franco et al, 2006;Nogueira et al, 2015;Osman et al, 2007;Zanette et al, 2011). In this regard, gills and digestive gland are the most commonly studied tissues: gills are considered the main interface between the organism, environment and, consequently, waterborne pollutants (Ravera, 2001), while digestive gland is known as a major detoxification site due to the higher expression of the mixed-function oxidase system (Livingstone and Farrar, 1984;Solé and Livingstone, 2005).…”

Section: Introductionmentioning

confidence: 99%

Gills as a glutathione-dependent metabolic barrier in Pacific oysters Crassostrea gigas : Absorption, metabolism and excretion of a model electrophile

et al. 2016

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The mercapturic acid pathway (MAP) is a major phase II detoxification route, comprising the conjugation of electrophilic substances to glutathione (GSH) in a reaction catalyzed by glutathione S-transferase (GST) enzymes. In mammals, GSH-conjugates are exported from cells, and the GSH-constituent amino acids (Glu/Gly) are subsequently removed by ectopeptidases. The resulting Cys-conjugates are reabsorbed and, finally, a mercapturic acid is generated through N-acetylation. This pathway, though very well characterized in mammals, is poorly studied in non-mammalian biological models, such as bivalve mollusks, which are key organisms in aquatic ecosystems, aquaculture activities and environmental studies. In the present work, the compound 1-chloro-2,4-dinitrobenzene (CDNB) was used as a model electrophile to study the MAP in Pacific oysters Crassostrea gigas. Animals were exposed to 10μM CDNB and MAP metabolites were followed over 24h in the seawater and in oyster tissues (gills, digestive gland and hemolymph). A rapid decay was detected for CDNB in the seawater (half-life 1.7h), and MAP metabolites peaked in oyster tissues as soon as 15min for the GSH-conjugate, 1h for the Cys-conjugate, and 4h for the final metabolite (mercapturic acid). Biokinetic modeling of the MAP supports the fast CDNB uptake and metabolism, and indicated that while gills are a key organ for absorption, initial biotransformation, and likely metabolite excretion, hemolymph is a possible milieu for metabolite transport along different tissues. CDNB-induced GSH depletion (4h) was followed by increased GST activity (24h) in the gills, but not in the digestive gland. Furthermore, the transcript levels of glutamate-cysteine ligase, coding for the rate limiting enzyme in GSH synthesis, and two phase II biotransformation genes (GSTpi and GSTo), presented a fast (4h) and robust (∼6-70 fold) increase in the gills. Waterborne exposure to electrophilic compounds affected gills, but not digestive gland, while intramuscular exposure was able to modulate biochemical parameters in both tissues. This study is the first evidence of a fully functional and interorgan MAP pathway in bivalves. Hemolymph was shown to be responsible for the metabolic interplay among tissues, and gills, acting as a powerful GSH-dependent metabolic barrier against waterborne electrophilic substances, possibly also participating in metabolite excretion into the sea water. Altogether, experimental and modeled data fully agree with the existence of a classical mechanism for phase II xenobiotic metabolism and excretion in bivalves.

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“…Processo de oxidação e redução é o processo biológico de maior abundância de trans-critos associados com essa função na brânquia (Figura 2.4), e ele está relacionado com várias funções comuns na célula (respiração celular, glicólise...), mas também está relacionado com atividades de biotransformação de xenobióticos e antioxidantes, uma vez que genes como CYP, SOD, GPx e CAT são atribuídos à essa categoria do GO. Trabalhos já mostraram que genes de CYP são expressos em brânquias de outros bivalves na presença de xenobióticos [1,15], e também existem trabalhos que mediram a atividade de enzimas antioxidantes em brânquias de P. perna e como elas respondem na presença de contaminantes [10,13,14] ou quando expostos ao ar [57]. Figura 2.4: Os 20 termos GO mais abundantes na categoria BP a nível de aminoácido.…”

Section: Anotação Funcionalunclassified

“…Uma das formas de se monitorar a qualidade dos ecossistemas marinhos é através do uso de biomarcadores. Em estudos com o P. perna, têm-se medido a atividade de enzimas de biotransformação de xenobióticos (glutationa-S-transferase (GST) e proteína de resistência a múltiplas drogas (MDR)) e antioxidantes (glutationa peroxidase (GPx), catalase (CAT) e superóxido dismutase (SOD)), como biomarcadores [10,13,14]. No entanto, a atividade enzimática apresenta respostas com um atraso maior, em relação ao momento que houve a exposição ao contaminante, do que a mudança do padrão de expressão gênica, como mostra um estudo de Lüchmann et al [1], onde ostras (Crassostrea brasiliana) foram expostas a fenantreno por 24h e apresentaram alterações no padrão de expressão de genes relacionados a biotransformação de HPAs, mas não houve uma alteração significativa na atividade dessas enzimas, o que nos mostra que alterações na expressão gênica são biomarcadores de resposta mais rápida do que atividade enzimática.…”

Section: Introductionunclassified

Análise do transcritoma do mexilhão marrom (Perna perna) sob contaminação por antraceno

Monteiro¹

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The brown mussel Perna perna (Linnaeus, 1758) helps the monitoring of chemical compounds in marine ecosystems. However its molecular mechanisms of detoxification and stress response remain unclear. Elucidating these mechanisms is crucial to understand the toxic effects of chemical pollutants and to develop biomarkers to assess marine ecosystems. In this study, P. perna individuals were exposed to anthracene (ANT) and its mRNA complement was sampled sequenced with Illumina technology. Chemical analysis of the soft tissue identified ANT concentrations 268-715 fold higher in the exposed group compared to controls, demonstrating that the exposure procedure was successfully accomplished. Transcriptome sequencing of P. perna generated 273.152.390 paired reads that were assembled in 231.728 contigs of average length 720 bp and N50 1.083 bp , which 66.563 contigs (28,7%) could be annotated using GenBank genes, Pfam domains, Gene Ontology (GO) terms and KEGG pathways. The results obtained from functional annotation suggest gills play a role in xenobiotics biotransformation, antioxidant response, signal transduction, innate immune response, and osmoregulation. It was possible to identify transcripts similar to genes related with biotransformation reactions of phases I, II and III, including CYPs and GSTs. Transcripts similar to CYPs and GSTs isoforms were highly expressed in the group exposed to ANT, however no CYP, GST, or even other genes related with biotransformation reactions were classified as differentially expressed. On the other hand, several hypothetical genes were differentially expressed, which suggests that P. perna uses unknown mechanisms of biotransformation to deal with ANT stress contamination. Immune related-genes were both up and downregulated, as was also observed for Perna viridis exposed to benzo(a)pyrene, suggesting that ANT promotes alteration in the immune response of P. perna.

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Biochemical responses in mussels Perna perna exposed to diesel B5

Cited by 17 publications

References 40 publications

Exposure to phenanthrene and depuration: Changes on gene transcription, enzymatic activity and lipid peroxidation in gill of scallops Nodipecten nodosus

Exposure to phenanthrene and depuration: Changes on gene transcription, enzymatic activity and lipid peroxidation in gill of scallops Nodipecten nodosus

Gills as a glutathione-dependent metabolic barrier in Pacific oysters Crassostrea gigas : Absorption, metabolism and excretion of a model electrophile

Análise do transcritoma do mexilhão marrom (Perna perna) sob contaminação por antraceno

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