Differential protein expression in the estuarine copepod Eurytemora affinis after diuron and alkylphenol exposures.

Proteomics was used in the calanoid copepod Eurytemora affinis for screening of protein expression modifications induced by organic contaminants. The copepods were exposed in a continuous flow-through system for 86 h to environmentally relevant concentrations of contaminants representative of the pollution in the Seine Estuary (Haute-Normandie, France; diuron, 500 ng L(-1) ; alkylphenol mixture, 1000 ng L(-1) ). Proteome analysis of whole-body copepod extracts by 2-dimensional gel electrophoresis revealed that the contaminants induced modifications in protein expression, with the highest quantitative variations occurring after diuron exposure. Specifically, 88 and 41 proteins were differentially expressed after diuron and alkylphenol treatments, respectively. After mass spectrometry analysis, 51 (diuron exposure) and 15 (alkylphenol exposure) proteins were identified. The identified proteins were potentially related to energy metabolism, cell growth, nervous signal conductivity, excitotoxicity, oxidative stress response, and antioxidant defense. The data suggest a massive general disturbance of physiological functions of E. affinis after diuron exposure, whereas alkylphenols induced an alteration of a few targeted physiological functions. The protein expression signatures identified after contaminant exposure deserve further investigation in terms of the development of novel potential biomarkers for water quality assessment. Environ Toxicol Chem 2016;35:1860-1871. © 2015 SETAC.

Molecular characterization and mRNA expression of grp78 and hsp90A in the estuarine copepod Eurytemora affinis.

The present study aimed to develop a method of quantification of heat shock protein transcript levels in the estuarine copepod Eurytemora affinis. For that, the full-length cDNA of the 78-kDa glucose-regulated protein (Ea-grp78) and the cytosolic 90-kDa heat shock protein (Ea-hsp90A) from this species have been cloned. These cDNA revealed, respectively, 2,370 and 2,299 bp with 1,971 and 2,124 bp open reading frames encoding 656 and 707 amino acids. Main features, sequence identities and phylogenetic analysis with other species were described. Then, the expression profiles were analysed using reverse transcription/real-time quantitative PCR method from copepods subjected to different thermic and osmotic stresses in laboratory, and from copepods directly sampled into the natural population of the Seine Estuary (France) along a salinity gradient. Thermic shock (7.5°C, 22.5°C and 30°C during 90 min) significantly induced increases of transcript quantities ranged between 1.7- and 19.7-fold the levels observed in control conditions (15°C). Hypo- and hyper-osmotic shocks (salinities of 1 and 30 during 90 min) caused a 2-fold induction of Ea-hsp90A transcript level in comparison to controls (salinity of 15) whereas no significant change was measured for Ea-grp78. On the other hand, similar expression profiles were observed for the two transcripts after 72 h of exposition to salinities of 1 and 25 with a significant 2-fold induction observed for the lower salinity. To finish, strong expression inductions of both Ea-grp78 and Ea-hsp90A genes were observed in field copepods sampled at low salinity during the campaigns of June 2009 and May 2010. These results tend to show that the low salinity and the increase of temperature seem to have a synergic effect on stress condition of copepods.