Analysis of Cyp2b1 gene expression in the rat liver and brain by multiplex PCR

Abstract

The CYP2B6 enzyme is involved in the metabolism of environmental toxins and endogenous compounds. The expression and activity of the cyp2b6 gene is important for the brain’s ability to detoxify harmful compounds. In rats, the Cyp2b1 gene is homologous to the human Cyp2b6 gene and has an isoform, Cyp2b2, with nearly the same sequence. Although the distinction between them is fundamental, most studies report global changes in CYP2B enzymes due to the complexity of the techniques to detect both genes separately. We instrumented a semi-quantitative technique combining multiplex end point RT-PCR with restriction enzyme assays, allowing us to establish a relatively simple, reproducible and affordable method to distinguish Cyp2b1 gene expression from Cyp2b2. Using this technique, we showed differential Cyp2b1 expression after phenobarbital and ethanol induction in the brain and liver and detected increased Cyp2b1 expression after ethanol treatment in the brain striatum that had not been detected before.