Department of Biological Science, College of Science, Sungkyunkwan University, Suwon 16419, South Korea.
Graduate School of Fisheries and Environmental Sciences, Nagasaki University, Nagasaki 852-8521, Japan; Organization for Marine Science and Technology, Nagasaki University, Nagasaki 852-8521, Japan.
Comp Biochem Physiol Part D Genomics Proteomics. 2019 Mar;29:185-192. doi: 10.1016/j.cbd.2018.12.001. Epub 2018 Dec 4.
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.
细胞色素 P450 酶（CYPs）是一个广泛存在于所有生物中的大型基因超家族，在解毒外来化合物和内源性化学物质方面起着关键作用。虽然水生无脊椎动物的 CYP 及其解毒机制已经有报道，但关于轮虫 Brachionus spp. 中 CYP 的种间比较及其解毒机制知之甚少。本研究的目的是鉴定轮虫 Brachionus rotundiformis（B. rotundiformis）中的全部 CYP，并将 B. rotundiformis-CYP 与先前报道的其他模式轮虫（B. koreanus、B. plicatilis 和 B. calyciflorus）中的 CYP 进行比较。为了验证该模型，轮虫，特别是 Brachionus rotundiformis 被暴露于各种浓度的 B[α]P 中，B[α]P 是一种广泛使用的多环芳烃外来化合物，并分析了对 B[α]P 的基因表达。在这里，通过计算机分析结果显示，轮虫 B. rotundiformis 中共有 26 种 CYP。基于系统发生分析，与三种轮虫物种 B. koreanus、B. plicatilis 和 B. calyciflorus 相比，B. rotundiformis 的 26 种 CYP 被分为五个不同的家族：2、3、4、线粒体和 46 家族。为了了解 26 种 B. rotundiformis-CYP 的解毒机制，我们研究了 26 种 CYP 的转录表达，发现 5 种 CYP（CYP3045A2、CYP3045B4、CYP3045C10、CYP3049A5 和 CYP3049E8）在 10 和 100μg B[α]P 作用下显著增加（P < 0.05）。此外，我们鉴定了芳香烃受体（AhR）和芳香烃受体核转位蛋白（ARNT），并观察到 B. rotundiformis-AhR 和 -ARNT 略有上调，表明这些 CYP 可能与解毒机制有关，并可作为 B. rotundiformis 中 B[α]P 的潜在分子生物标志物。总的来说，这项研究将有助于我们扩大对轮虫中与 AhR 信号通路相关的解毒机制的无脊椎动物 CYP 的认识。
