Dept. of Environmental Toxicology, Evolutionary Biology Centre, Uppsala University, Norbyvägen 18A, 75236 Uppsala, Sweden.
Toxicol Appl Pharmacol. 2011 Jan 15;250(2):170-83. doi: 10.1016/j.taap.2010.10.010. Epub 2010 Oct 18.
The Xenopus tropicalis genome shows a single gene in each of the four cytochrome P450 1 (CYP1) subfamilies that occur in vertebrates, designated as CYP1A, CYP1B1, CYP1C1, and CYP1D1. We cloned the cDNAs of these genes and examined their expression in untreated tadpoles and in tadpoles exposed to waterborne aryl hydrocarbon receptor agonists, 3,3',4,4',5-pentachlorobiphenyl (PCB126), β-naphthoflavone (βNF), or indigo. We also examined the effects of PCB126 on expression of genes involved in stress response, cell proliferation, thyroid homeostasis, and prostaglandin synthesis. PCB126 induced CYP1A, CYP1B1, and CYP1C1 but had little effect on CYP1D1 (77-, 1.7-, 4.6- and 1.4-fold induction versus the control, respectively). βNF induced CYP1A and CYP1C1 (26- and 2.5-fold), while, under conditions used, indigo tended to induce only CYP1A (1.9-fold). The extent of CYP1 induction by PCB126 and βNF was positively correlated to the number of putative dioxin response elements 0-20 kb upstream of the start codons. No morphological effect was observed in tadpoles exposed to 1 nM-10 μM PCB126 at two days post-fertilization (dpf) and screened 20 days later. However, in 14-dpf tadpoles a slight up-regulation of the genes for PCNA, transthyretin, HSC70, Cu-Zn SOD, and Cox-2 was observed two days after exposure to 1 μM PCB126. This study of the full suite of CYP1 genes in an amphibian species reveals gene- and AHR agonist-specific differences in response, as well as a much lower sensitivity to CYP1 induction and short-term toxicity by PCB126 compared with in fish larvae. The single genes in each CYP1 subfamily may make X. tropicalis a useful model for mechanistic studies of CYP1 functions.
爪蟾基因组在脊椎动物发生的四个细胞色素 P450 1(CYP1)亚家族中的每一个都显示出一个基因,分别命名为 CYP1A、CYP1B1、CYP1C1 和 CYP1D1。我们克隆了这些基因的 cDNA,并研究了它们在未处理的蝌蚪和暴露于水传播的芳烃受体激动剂 3,3',4,4',5-五氯联苯(PCB126)、β-萘黄酮(βNF)或靛蓝中的表达。我们还研究了 PCB126 对参与应激反应、细胞增殖、甲状腺稳态和前列腺素合成的基因表达的影响。PCB126 诱导 CYP1A、CYP1B1 和 CYP1C1,但对 CYP1D1 几乎没有影响(分别为对照的 77、1.7、4.6 和 1.4 倍诱导)。βNF 诱导 CYP1A 和 CYP1C1(26 和 2.5 倍),而在使用的条件下,靛蓝似乎只诱导 CYP1A(1.9 倍)。PCB126 和 βNF 诱导 CYP1 的程度与起始密码子上游 0-20kb 处的推定二恶英反应元件的数量呈正相关。在受精后两天(dpf)暴露于 1 nM-10 μM PCB126 并在 20 天后筛选的蝌蚪中未观察到形态学效应。然而,在 14-dpf 蝌蚪中,暴露于 1 μM PCB126 两天后,PCNA、转甲状腺素蛋白、HSC70、Cu-Zn SOD 和 Cox-2 的基因表达略有上调。这项对两栖动物物种中全套 CYP1 基因的研究揭示了基因和 AHR 激动剂特异性的反应差异,以及与鱼类幼虫相比,PCB126 对 CYP1 诱导和短期毒性的敏感性要低得多。每个 CYP1 亚家族中的单个基因可能使爪蟾成为 CYP1 功能的机制研究的有用模型。