cDNA fingerprint from the hepatopancreatic glands of pond snails (Lymnaea stagnalis ) exposed to benzo( a)pyrene

Abstract

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