Respiration disruption and detoxification at the protein expression levels in the Pacific oyster (Crassostrea gigas) under zinc exposure

The Pacific oyster (Crassostrea gigas) can accumulate high levels of zinc (Zn) without obvious toxicity, but the related molecular mechanisms are largely unknown. In the present study, C. gigas were exposed to excess Zn for 9 days and the differentially expressed proteins (DEPs) were examined using the isobaric tags for relative and absolute quantitation (iTRAQ) method. In total, 2667 proteins containing at least two peptides and detected in both replicates were used for proteomic analysis. Among these DEPs, 332 were up-regulated and 282 were down-regulated. KEGG enrichment analysis of DEPs revealed that Zn exposure mainly distrubed 'tricarboxylic acid (TCA) cycle', 'electron transport chain (ETC)' and 'glutathione (GSH) metabolism' processes in oysters. Further key protein expressions enriched in these metabolism pathways were analyzed. In TCA cycle, Zn inhibited the Fe-containing protein expressions, which may lead to the accumulation of succinate and induce anaerobiosis. In ETC metabolism process, Zn inhibited ETC complex protein expressions, including complex I-IV, which may affect the electron transport process. Furthermore, Zn induced phytochelatin (PC) and glutathione peroxidase (GPX) expression in GSH catabolism. The proteins play important roles in Zn detoxification and ROS elimination process. The transcriptional expressions of genes encoding these proteins were observed using real-time PCR analysis, and there was good consistency between these two datasets. Overall, we provide direct evidence for Zn toxicity and detoxification mechanisms at protein level.