Expression and inducibility of aryl hydrocarbon receptor pathway genes in wild-caught killifish (Fundulus heteroclitus) with different contaminant-exposure histories
Expression and inducibility of aryl hydrocarbon receptor pathway genes in wild-caught killifish (Fundulus heteroclitus) with different contaminant-exposure histories
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DOI：
10.1897/02-495
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发表时间：
2003-10-01
影响因子：
4.1
通讯作者：
Di Giulio, RT
中科院分区：
文献类型：
--
作者：
Meyer, JN;Wassenberg, DM;Di Giulio, RT
从伊丽莎白河（弗吉尼亚州，美国）的污染场地野生捕捞的鳉鱼是难诱导细胞色素P4501 A（CYP 1A，作为催化活性和免疫检测CYP 1A蛋白）暴露后，典型的芳烃受体（AHR）激动剂，已报告的鱼类从其他网站高度污染这些化合物。在试图了解缺乏诱导的CYP 1A蛋白的表达和活性在伊丽莎白河鳉的分子基础上，我们分析了CYP 1A和其他四个成员的AHR信号转导通路：AHR 1，AHR 2，AHR阻遏物（AHRR），和AHR核转位（ARNT）的表达。通过循环优化逆转录酶-聚合酶链反应（RT-PCR）分析从伊丽莎白河和King's Creek（VA，USA）（参考地点）的鳉鱼肝脏中提取的信使核糖核酸（mRNA），在注射β-萘酚酮（BNF，一种AHR激动剂）或玉米油（载体对照）后36 h，测定基因表达。肝脏CYP 1A mRNA在King's Creek Killifish中是可诱导的。然而，在伊丽莎白河鳉鱼中，没有观察到CYP 1A mRNA的诱导，证实并扩展了先前的结果，即在该种群中没有CYP 1A蛋白或催化活性的诱导。同样，AHRR和AHR 2 mRNA水平诱导BNF在国王溪，但不是伊丽莎白河？在AHR 1或ARNT mRNA的表达中未观察到群体或治疗相关差异。结果表明，在伊丽莎白河？AHRR，AHR2）。然而，AHR 1，AHR 2和AHRR的表达在媒介物处理的鱼没有伊丽莎白河和国王溪？
Wildcaught killifish from a contaminated site on the Elizabeth River (VA, USA) are refractory to induction of cytochrome P4501A (CYP1A, measured as catalytic activity and immunodetectable CYP1A protein) after exposure to typical aryl hydrocarbon receptor (AHR) agonists, as has been reported for fish from other sites highly contaminated with these compounds. In an attempt to understand the molecular basis for the lack of inducibility of CYP1A protein expression and activity in Elizabeth River killifish, we analyzed the expression of CYP1A and four other members of the AHR signal transduction pathway: AHR1, AHR2, AHR repressor (AHRR), and AHR nuclear translocator (ARNT). Gene expression was measured by cycle-optimized reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of messenger ribonucleic acid (mRNA) extracted from livers of killifish from the Elizabeth River and King's Creek (VA, USA) (reference site), 36 h after injection with beta-naphthoflavone (BNF, an AHR agonist) or corn oil (carrier control). Hepatic CYP1A mRNA was inducible in King's Creek killifish. However, in Elizabeth River killifish, no induction of CYP1A mRNA was observed, confirming and extending previous results showing no induction of CYP1A protein or catalytic activity in this population. Similarly, AHRR and AHR2 mRNA levels were induced by BNF in King's Creek but not Elizabeth River killifish. No population or treatment-related differences were observed in expression of AHR1 or ARNT mRNAs. The results reveal in Elizabeth River killifish a consistent lack of inducibility of genes that are normally inducible by AHR agonists (CYP1A. AHRR, AHR2). However, the expression of AHR1, AHR2, and AHRR in vehicle-treated fish did not differ between Elizabeth River and King's Creek killifish, suggesting that altered constitutive expression of AHRs or AHRR does not underlie the refractory CYP1A phenotype in Elizabeth River killifish.