Effects Of Exposure To Nonylphenol On Expression Of Heat Shock Protein Of Zebrafish

Heat shock proteins(HSPs)is a highly conserved evolutionary and encoded by heat shock genes of stress proteins in all living organisms which is induced by harmful factors in the living environment such as temperature mutation,pathogen infection,physical and chemical factors.Many heat shock proteins have molecular chaperone activity.They played a very important role in protecting the normal organism cell,fighting cell decline,repairing and folding damaged proteins and disease warning and so on.HSP70 gene family and HSP90 gene family are the most important two families,which are early warning indicators for ecological health situation of the water environment.Due to the aquatic animal zebrafish has the advantage of the strong reproductive ability,mass culture easily and high sensitivity,therefore this research issue uses them as the experiment materials to use the technology of real-time fluorescence quantitative PCR to detect expression of the three heat shock genes(HSP70,HSP90 a,HSP90b)in the tissue of liver,brain and muscle by way of the direct exposure and food chain indirectly exposure.The main conclusions are as follows:(1)Experimental analysis results showed that NP exposure directly induced HSP70 gene,HSP90 a gene and HSP90 b gene specific expression in zebrafish,the expression discipline of three genes in three tissue(liver,brain and muscle)were consistent,within a certain time range,have shown to increase to a maximum value and then decreased,with the continued extension of the processing time,the recovered gradually;When the expression of HSP70 gene in the liver,brain and muscle expression peaked at 8d,when the expression of HSP90 a gene in liver and brain peaked at 8d,it did at 24 d in muscle,and when the expression of HSP90 b gene in liver and brain peaked at 8d,it did at 16 d in muscle.(2)The expression discipline by way of the food chain indirectly expose experiment treated with NP were consistent with the relative expression of the heat shock proteins gene by way of directly expose zebrafish experiment treated with NP were consistent with the condition of NP direct exposure to zebrafish.The relative expression was higher than the relative expression of the heat shock proteins gene by way of directly expose zebrafish experiment treated with NP,So we inferred that this phenomenon may be related to different content of NP accumulation in zebrafish,which is consistent to the result that the low concentration NP induced heat shock proteins expression was higher than the heat shock proteins expression induced by the high concentration of NP.(3)With ?-actin as the reference gene to establish the detection method of the real-time fluorescence quantitative PCR technology for HSP70 mRNA,HSP90 a mRNA and HSP90 b mRNA.The analysis result showed that the three genes were sensitive to nonylphenol,especially HSP70 mRNA was most sensitive in three genes,it can be used as an early warning biomarker for pollutant.