| As a typical organophosphorus pollutant,tris(1,3-dichloroisopropyl)phosphate(TDCIPP)has been widely detected in aquatic environment.Previous studies showed that protein phosphorylation might be a vital way of TDCIPP to exert multiple toxic effects.However,there is a lack of investigations on how TDCIPP affected protein phosphorylation.In order to explore the toxicological effects and mechanisms of TDCIPP,proteomic and phosphoproteomic analyses together with evaluation of histopathological changes and enzymatic activities were conducted in marine oysters Crassostrea gigas treated with 0.5,5 and 50 μg/L TDCIPP for 28 days.Results showed that(1)TDCIPP was significantly enriched in 5 and 50 μg/L TDCIPP-treated oysters.Cytochromes P450 might be involved in the phase I metabolism of TDCIPP in 50 μg/L TDCIPP-treated group.(2)TDCIPP induced neurotoxicity in oysters.ACh E activities were inhibited by low and moderate doses(0.5 and 5 μg/L).Higher ACh E activity caused by high dose(50 μg/L)of TDCIPP could be explained by the overcompensation.(3)The atrophic digestive cells and thinner epithelium were observed after TDCIPP treatment.Even shedding of partial digestive cells occurred was observed.(4)Proteomic and phosphoproteomic analysis showed that a total of 551 differentially expressed proteins and 152 differentially phosphorylated proteins were identified in 50μg/L TDCIPP-treated group.Bioinformatics analyses showed that TDCIPP dysregulated transcription,energy metabolism,and apoptosis and cell proliferation by phosphorylating pivotal proteins and enzymes or signaling pathways.TDCIPP could promote apoptosis by increasing caspase-9 activity,and promoted proliferation of digestive cells.TDCIPP could accelerate transcription initiation and pre-m RNA processing for regulation of life activities.TDCIPP impeded aerobic respiration and disrupted energy metabolism by inhibition of MDH and PDH via phosphorylation.This study predicted the association between toxicological effects caused by TDCIPP treatments and protein phosphorylation or dephosphorylation by analyzing proteome and phosphoproteome of oysters,and revealed the toxicological effects and mechanisms of TDCIPP in marine bivalve C.gigas,which provides the theoretical support for risk assessment of organophosphorus pollution. |
