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Identification and Expression of Multiple CYP1-like and CYP3-like Genes in the Bivalve Mollusk Mytilus edulis

机译:鉴定和表达二级CYP1样MOLLUSK Mytilus Edulis中的多种CYP1样和CYP3样基因

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摘要

Various sequencing projects over the last several years have aided the discovery of previously uncharacterized invertebrate sequences, including new cytochrome P450 genes (CYPs). Here we present data on the identification and characterization of two CYP1-like and three CYP3-like genes from the bivalve mollusk Mytilus edulis, and assess their potential as biomarkers based on their responses to several known vertebrate aryl hydrocarbon receptor (AHR) agonists. Quantitative real-time PCR was used to measure CYP transcript levels in digestive gland, labial palps, adductor muscle, gill, foot, and different regions of the mantle. Levels of both CYP1-like genes were highest in digestive gland, whereas labial palps had the highest expression levels of the three CYP3-like genes followed by digestive gland and outer margin of the mantle. Mussels were exposed by injection to the AHR agonists, β-naphthoflavone (BNF; 25 μg.g−1), 3,3′,4,4′,5-polychlorinated biphenyl (PCB126; 2 μg.g−1), or 6-formylindolo[3,2-b]carbazole (FICZ; 0.1 μg.g−1), or to Aroclor 1254 (a mixture of PCBs; 50 μg.g−1) for 24 hours, followed by CYP expression analysis. There was no statistically significant change in expression of either of the CYP1-like genes after exposure to the various AHR agonists. The CYP3-like-1 gene was significantly up-regulated by BNF in gill tissues and the CYP3-like-2 gene was up-regulated in digestive gland by PCB126 and in gill tissue by BNF. These results suggest that distinct mechanisms of CYP gene activation could be present in M. edulis, although the importance of the CYP1-like and CYP3-like genes for xenobiotic and endogenous lipids biotransformation requires additional investigation.
机译:在过去的几年中,各种测序项目帮助发现了以前未表征的无脊椎动物序列,包括新的细胞色素P450基因(CYP)。在这里,我们介绍从双壳软体动物Mytilus edulis鉴定和表征两个CYP1样和三个CYP3样基因的数据,并根据它们对几种已知的脊椎动物芳烃受体(AHR)激动剂的反应,评估其作为生物标志物的潜力。实时荧光定量PCR用于测量消化腺,阴唇掌,内收肌,g,足和地幔不同区域的CYP转录水平。两种CYP1样基因的水平在消化腺中最高,而唇瓣在三个CYP3样基因中的表达水平最高,其次是消化腺和地幔的外缘。通过注射将贻贝暴露于AHR激动剂β-萘黄酮(BNF; 25μg.g -1 ),3,3',4,4',5-多氯联苯(PCB126; 2μg .g −1 )或6-甲酰基吲哚[3,2-b]咔唑(FICZ; 0.1μg.g -1 ),或Aroclor 1254(混合物多氯联苯; 50μg.g -1 )放置24小时,然后进行CYP表达分析。暴露于各种AHR激动剂后,任一CYP1样基因的表达均无统计学上的显着变化。 CYP3-like-1基因在g组织中被BNF显着上调,而CYP3-like-2基因在PCB126中的消化腺中和在BNF中在g组织中被上调。这些结果表明,CYP基因激活的独特机制可能存在于蓝藻中,尽管CYP1类和CYP3类基因对于异种和内源性脂质生物转化的重要性还需要进一步研究。

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