Identification of the full 26 cytochrome P450 (CYP) genes and analysis of their expression in response to benzo[α]pyrene in the marine rotifer Brachionus rotundiformis

Abstract

Cytochrome P450s (CYPs) are a large gene superfamily that are found in all living organisms. CYPs have a key role in detoxification of xenobiotics and endogenous chemicals. Although aquatic invertebrate CYPs and their detoxification mechanisms have been reported, little is known about interspecific comparison of CYPs and their detoxification mechanism in the rotifer Brachionus spp. The aim of this study was to identify the entire CYPs in the rotifer Brachionus rotundiformis (B. rotundiformis) and compare B. rotundiformis-CYPs to the previously reported CYPs in other model Brachionus spp. (B. koreanus, B. plicatilis, and B. calyciflorus). To validate the model, the rotifer, specifically Brachionus rotundiformis was exposed to various concentrations of B[α]P, which is widely used PAH xenobiotic, and analyzed gene expression in response to B[α]P. Here, in silico analysis results showed the total of 26 CYPs from the rotifer B. rotundiformis. Based on the phylogenetic analysis, the 26 B. rotundiformis-CYPs were separated into five different clans: 2, 3, 4, mitochondrial, and 46 clans in comparison to three rotifers species, B. koreanus, B. plicatilis, and B. calyciflorus. To understand the detoxification mechanisms of 26 B. rotundiformis-CYPs, we investigated transcriptional expression of 26 CYPs and found that five CYPs (CYP3045A2, CYP3045B4, CYP3045C10, CYP3049A5, and CYP3049E8) were significantly increased (P < 0.05) in response to 10 and 100 μg B[α]P. In addition, we identified the aryl hydrocarbon receptor (AhR) and aryl hydrocarbon receptor nuclear translocator (ARNT) and observed slight up-regulation of B. rotundiformis-AhR and -ARNT, indicating that these CYPs are likely associated with detoxification mechanism and could be used as potential molecular biomarkers of B[α]P in B. rotundiformis. Overall, this study will be helpful for expanding our knowledge of invertebrate CYPs on detoxification mechanisms associated with AhR signaling pathway in rotifers.