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  • Changes in protein expression of pacific oyster Crassostrea gigas exposed in situ to urban sewage.

Changes in protein expression of pacific oyster Crassostrea gigas exposed in situ to urban sewage.

Environmental science and pollution research international (2014-11-16)
Fabrício Flores-Nunes, Tânia Gomes, Rui Company, Roberta R M Moraes, Silvio T Sasaki, Satie Taniguchi, Márcia C Bicego, Cláudio M R Melo, Afonso C D Bainy, Maria J Bebianno
ABSTRACT

The composition and concentration of substances in urban effluents are complex and difficult to measure. These contaminants elicit biological responses in the exposed organisms. Proteomic analysis is a powerful tool in environmental toxicology by evidencing alterations in protein expression due to exposure to contaminants and by providing a useful framework for the development of new potential biomarkers. The aim of this study was to determine changes in protein expression signatures (PES) in the digestive gland of oysters Crassostrea gigas transplanted to two farming areas (LIS and RIB) and to one area contaminated by sanitary sewage (BUC) after 14 days of exposure. This species is one of the most cultivated molluscs in the world. The identified proteins are related to the cytoskeleton (CKAP5 and ACT2), ubiquitination pathway conjugation (UBE3C), G protein-coupled receptor and signal transduction (SVEP1), and cell cycle/division (CCNB3). CKAP5 showed higher expression in oysters kept at BUC in comparison with those kept at the farming areas, while ACT2, UBE3C, SVEP1, and CCNB3 were suppressed. The results suggest that these changes might lead to DNA damage, apoptosis, and interference with the immune system in oyster C. gigas exposed to sewage and give initial information on PES of C. gigas exposed to sanitary sewage, which can subsequently be useful in the development of more sensitive tools for biomonitoring coastal areas, particularly those devoted mainly to oyster farming activities.

MATERIALS
Product Number
Brand
Product Description

SAFC
HEPES
Sigma-Aldrich
Phenylmethanesulfonyl fluoride, ≥99.0% (T)
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Chlorotrimethylsilane, produced by Wacker Chemie AG, Burghausen, Germany, ≥99.0% (GC)
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HEPES, BioUltra, for molecular biology, ≥99.5% (T)
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Chlorotrimethylsilane, purified by redistillation, ≥99%
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Phenylmethanesulfonyl fluoride, ≥98.5% (GC)
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HEPES, BioXtra, pH 5.0-6.5 (1 M in H2O), ≥99.5% (titration)
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HEPES, BioXtra, suitable for mouse embryo cell culture, ≥99.5% (titration)
Supelco
N,O-Bis(trimethylsilyl)trifluoroacetamide with trimethylchlorosilane, with 1% trimethylchlorosilane, for GC derivatization, LiChropur
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HEPES, BioPerformance Certified, ≥99.5% (titration), suitable for cell culture
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HEPES, ≥99.5% (titration)
SAFC
HEPES
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Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
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HEPES, anhydrous, free-flowing, Redi-Dri, ≥99.5%
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Chlorotrimethylsilane, ≥98.0% (GC)
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Sucrose, SAJ first grade
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Sucrose, JIS special grade
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HEPES buffer solution, 1 M in H2O
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N,O-Bis(trimethylsilyl)trifluoroacetamide, ≥99%
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Chlorotrimethylsilane solution, 1.0 M in THF
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Sucrose, BioUltra, for molecular biology, ≥99.5% (HPLC)
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Sucrose, ≥99.5% (GC), BioReagent, suitable for cell culture, suitable for insect cell culture
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Sucrose, for molecular biology, ≥99.5% (GC)
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Sucrose, ≥99.5% (GC)
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Sucrose, ACS reagent
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Sucrose, Grade I, ≥99% (GC), suitable for plant cell culture
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Sucrose, ≥99.5% (GC), BioXtra
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Sucrose, puriss., meets analytical specification of Ph. Eur., BP, NF