Differential Proteome Of Liver And Brain Tissues In Paralichthys Olivaceus Under The Stress Of Cadmium And Methyl Parathion

Methods of water pollution detection mainly contain physical and chemical monitoring and biological monitoring. The fore could be well competent for the instantaneous and local pollution, and the latter meets the shortfall and play an important role in the objectively detection the harmful degree of the pollutants to the organism. In the degree paper, the writer choosed Paralichthys olivaceus (P. olivaceus) as research animal and different-display proteomics as main separated and analytical technologies to identify the different display proteins in the liver proteomic under the cadmium stress and in the brain proteomic under the Methyl parathion stress, respectively. Using Introduced Inductively Coupled Plasma Mass Spectroscopy (ICP-MS) to estimate the concentration of the cadmium in the several tissues of the P. olivaceus which had been feeding in the cadmium polluted seawater for 96 h, the result showed that the cadmium was obviously enriched which was up to 6-fold in the liver. It was indicated that cadmium may induce the most different proteins in the liver, and the most obvious trend change, which was likely to be optimum tissue for the different-display proteomics analysis and for the seeking of proteins acting as the indicators of the pollution degree and harmfulness under cadmium stress.The technology of differential centrifugation for liver proteomic preseparation which efficiently optimized the extraction strategy of the low-abundance proteins, was meaningful for increasing the categories of indicators and the detect sensitivity. Spectrum of two-dimensional electrophoresis (2D-PAGE) showed up to 1,900 spots of total protein in liver preseparated by differential centrifugation. Compared analysis by the proteomic software, 51 differential proteins were determined, which were obtained for further peptide mass fingerprinting (PMF) analysis and database research. Proteins relative to cadmium toxicity were detected, such as heat shock proteins, catalase and so on. It is suggested that the different display proteins are meaningful as the protein indicators for the monitoring and estimation of cadmium pollution degree and harmfulness, and for the cadmium toxicology and passway research. Two of the main strategies for organic phosphorus pollution detecting are residue testing and enzymes activity assay in the organism. Zebrafish (classical biological model) was chosen as the research animal for the simulated experiment. Studies on the residues and the trend change by gas chromatography-mass Spectrometry (GC-MS), and the activity trend of SOD, CAT, AchE by activity assay in the different tissues under the Methyl parathion (MP) induced. The result showed that MP mostly enriched in the liver, and AchE activity was consistently suppressed, furthermore, the purpose of the simulated experiment was to set up a research model for seeking the proteins as the indicators of the pollution degree and harmfulness under MP stress in P. olivaceus.In the case of residues analysis in liver and AchE enzyme activity assay in brain of P. olivaceus, under MP stress, the results were identical to that in the simulated experiment. 17 different expression proteins in brain of P. olivaceus under the MP stress were obtained by the utilization of different-display proteomics, containing heat shock proteins, glutathione S-transferase and so on. These proteins were meaningful findings not only for the monitoring proteins as the indicators of the pollution degree and harmfulness, but also for the MP toxicological mechanism and passway research, with the significant scientific research value and potential application value.