Menzel R, Bogaert T, Achazi R
Department of Biology, Institute of Ecotoxicology & Biochemistry, Free University Berlin, Ehrenbergstrasse 26-28, 14195 Berlin, Germany.
Arch Biochem Biophys. 2001 Nov 15;395(2):158-68. doi: 10.1006/abbi.2001.2568.
The soil nematode Caenorhabditis elegans is one of the simplest animals having the status of a laboratory model. Its genome contains 80 cytochrome P450 genes (CYP). In order to study CYP gene expression in C. elegans mixed stages and synchronized hermaphrodites were exposed to 18 known xenobiotic cytochrome P450 inducers. Messenger RNA expression was detected by DNA arrays and semiquantitative RT-PCR. Using subfamily-specific primers, a pooled set of exon-rich CYP fragments could be amplified. In this way it was possible to systematically check the influence of different inducers on CYP expression at the same time. The well-known CYP1A inducers beta-naphthoflavone, PCB52, and lansoprazol were the most active and in particular they strongly induced almost all CYP35 isoforms. A few number of further CYP forms were found to be inducible by other xenobiotics like phenobarbital, atrazine, and clofibrate. In addition, a transgenic C. elegans line expressing GFP under control of the CYP35A2 promoter showed a strong induction of the fusion by beta-naphthoflavone in the intestine.
土壤线虫秀丽隐杆线虫是具有实验室模型地位的最简单动物之一。其基因组包含80个细胞色素P450基因(CYP)。为了研究秀丽隐杆线虫混合发育阶段的CYP基因表达情况,将同步化的雌雄同体线虫暴露于18种已知的外源性细胞色素P450诱导剂中。通过DNA芯片和半定量RT-PCR检测信使RNA表达。使用亚家族特异性引物,可以扩增出一组富集外显子的CYP片段。通过这种方式,可以同时系统地检测不同诱导剂对CYP表达的影响。著名的CYP1A诱导剂β-萘黄酮、多氯联苯52和兰索拉唑活性最强,尤其是它们强烈诱导了几乎所有的CYP35亚型。还发现一些其他的CYP形式可被苯巴比妥、阿特拉津和氯贝丁酯等其他外源性物质诱导。此外,在CYP35A2启动子控制下表达绿色荧光蛋白(GFP)的转基因秀丽隐杆线虫品系在肠道中显示出β-萘黄酮对融合蛋白的强烈诱导作用。