Research Note: Quantification of increase in hepatic CYP1A1 mRNA by real-time RT-PCR after exposure of the European eel (Anguilla anguilla) to a diesel oil water soluble fraction
Résumé
Editors' note: Research Notes is a new section now being added to the EBI format. Its purpose is to provide a forum for the publication of short research communications that provide important information but, due to their restricted focus, are not formatted in the manner of full research papers. All papers published under this heading will be peer reviewed. In order to follow the effects of pollutants such as polycyclic aromatic hydrocarbons (PAHs), polychlorobenzenes (PCBs), and dioxins upon the European eel, we have developed an assay that quantifies hepatic cytochrome P4501A1 (CYP1A1) mRNA levels using real-time reverse transcription polymerase chain reaction (real-time RT-PCR). In this study, European eels were exposed to a diesel water soluble fraction (DWSF) in order to mimic environmental exposure to a mixture of organic pollutants. CYP1A1 mRNA levels were measured after 3, 6, 12, 24, 48, 72 and 156 h of exposure. The 7-ethoxyresorufin O-deethylase (EROD) activity was measured in parallel to confirm the impact of the DWSF exposure at the protein activity level. DWSF exposure led to an increase in both CYP1A1 mRNA level and EROD activity. The increase in CYP1A1 mRNA level occurred very rapidly and was significant throughout the experiment, whereas EROD activity was increased only after 48 h of exposure. Quantification of CYP1A1 mRNA levels by real-time RT-PCR can therefore be used as a reliable biomarker of exposure of the European eel to CYP1A1 inducers in natural environments. This modern analytical method has the advantage of being appropriate for small quantities of tissue and is easy to perform at the same time as other biomarkers. Copyright © Taylor & Francis Group, LLC.