暴露于苯并(a)芘的塘螺肝胰腺cDNA指纹图谱

M. Woźny, M. Kowal, S. Ciesielski, M. Florczyk, R. Wiśniewski, E. Malicka, P. Brzuzan
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摘要

鉴定可能作为普通无脊椎动物(如淡水蜗牛)多环芳烃暴露的生物标志物的差异表达基因,将是对水生环境毒理学和生物监测感兴趣的研究人员的宝贵资源。因此,本研究的目的是研究水生苯并[a]芘(B[a]P)暴露对塘螺肝胰脏mRNA表达的影响。为此,采用50µM B[a]P溶液对池塘蜗牛(L. stagnation)成熟个体进行了短时间36h的静态暴露试验。采用差分显示PCR (DD-PCR)技术对暴露和未暴露的钉螺组织中差异表达的基因进行了独特的cDNA指纹图谱分析。为了评估分离的cDNA扩增子(ESTs)的假定身份,进行BLAST查询以发现其核苷酸序列的相似性。采用Real-Time qPCR分析验证DDPCR表达谱。最后,进行了另一项独立暴露研究,包括更高剂量的B[a]P(100µM),以验证所选ESTs的表达。爆炸
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cDNA fingerprint from the hepatopancreatic glands of pond snails (Lymnaea stagnalis ) exposed to benzo( a)pyrene
Identification of differentially expressed genes that could be potentially used as biomarkers of PAH exposure of common invertebrate animal (like freshwater snail) would be a valuable resource for investigators interested in toxicology and biomonitoring of aquatic environments. Therefore, the aim of this research was to investigate effects of waterborne benzo[ a]pyrene (B[ a]P) exposure on mRNA expression in the pond snail’s (Lymnaea stagnalis ) hepatopancreatic gland. Toward this end, mature individuals of pond snail ( L. stagnalis ) were treated with 50µM B[ a]P solution in a short 36h static exposure test. Differential Display PCR (DD-PCR) was used to generate a unique cDNA fingerprint of genes that were differentially expressed in the tissues of exposed and unexposed snails. To assess the putative identity of the isolated cDNA amplicons (ESTs), BLAST queries were performed to find similarities in their nucleotide sequence. Real-Time qPCR analysis was used to verify the DDPCR expression profile. Finally, an additional independent exposure study, including higher dose of B[ a]P (100µM), was conducted to validate the expression of selected ESTs. BLAST
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