Marine Biotech. Fellowship: Molecular Characterization of Cytochrome P450(s) in a Marine Fish: Applications to Environmental and Ecological Questions

海洋生物技术。

基本信息

项目摘要

The objectives of this project are to clone and characterize induced cytochrome P450 cDNA(s) from hepatic tissue of the fish, Chaetodon capistratus, and develop quantitative assays for cytochrome P450 mRNA and enzyme levels. Induction of cytochrome P450 gene expression will be characterized in C. capistratus using two model substrates: 1) beta-naphthoflavone (beta-NF), a 3- methylcholanthrene type xenobiotic and 2) curcuhydroquinone, and ichthyodeterrent allelochemical from the gorgonian coral, Pseudopterogorgia rigida. Characterization utilizing the first substrate will address the potential use of C. capistratus as an environmental biomonitor in coral reef environments. Characterization utilizing the second substrate will address the effect of an ichthyodeterrent allelochemical on P450 induction. Taken together, the evolutionary relationship between detoxification of these two classes of foreign compounds can be addressed. Furthermore, the potentially confounding effect of dietary allelochemicals on xenobiotically induced P450 will be investigated. Field sampling sites with and without recent polycyclic aromatic hydrocarbon (PAH) exposure will then test the feasibility of using C. capistratus cytochrome induction as a biomonitor of environmental contamination. Coral reefs are ecologically and economically important environments which are increasingly being subjected to detrimental anthropogenic influences, but effective monitoring mechanisms are lacking. Chronic, sublethal effects of pollution due to contamination by PAH and synthetic organic chemicals, though not readily apparent, can become lethal. The ability to detect low levels of contaminants may be crucial for preventing long-term damage. There are presently no molecular studies which have characterized a PAH-induced P450 in a tropical marine organism. Characterization of C. capistratus P450 will not only add to our knowledge of the diversity of this biologically important class of enzymes, but more importantly will provide a molecular probe with which to assess questions of ecological as well as environmental significance with great sensitivity. In particular, is cytochrome P450 responsible for the ability of C. capistratus to feed on gorgonian corals which are otherwise defended by ichthyodeterrent allelochemicals. This finding has potentially broad significance for understanding patterns of predation and herbivory in the marine environment. Molecular characterization of C. capistratus P450 and the development of molecular probes to test the effects of chemicals at sublethal levels thus has applicability to important environmental and ecological questions.
该项目的目标是克隆和表征 从鱼的肝组织中诱导细胞色素P450 cDNA, 头状蝴蝶鱼,并开发定量分析, 细胞色素P450 mRNA和酶水平。 诱导细胞色素 P450基因表达将在C. capistratus使用 两种模型底物:1)β-萘甲酮(β-NF),3- 甲基胆蒽型异生物质和2)姜黄氢醌,和 柳珊瑚中的鱼类威慑化感物质, 硬拟柳珊瑚 利用第一个特征 衬底将解决C的潜在用途。capistratus作为一个 珊瑚礁环境生物监测。 利用第二衬底的表征将解决 一种鱼类忌避化感物质对P450诱导作用 综合来看, 这两类外来化合物的解毒可以 处理。 此外,潜在的混杂效应 膳食化感物质对外源性诱导的P450的影响, 研究了 有和没有最近 多环芳烃(PAH)暴露将测试 利用C. capistratus细胞色素诱导作为一种 环境污染的生物监测。 珊瑚礁在生态和经济上都很重要 越来越多的环境受到有害的影响, 人为影响,但有效的监测机制, 缺乏 污染造成的慢性、亚致死影响, 多环芳烃和合成有机化学品的污染,虽然不是 很明显,会变得致命 低检测能力 污染物的水平可能是至关重要的, 损害 目前还没有分子研究, 表征了热带海洋生物中PAH诱导的P450。 对C. Capistratus P450不仅将增加我们的 了解这一生物学上重要的物种的多样性, 酶,但更重要的是将提供一种分子探针, 评估生态和环境问题, 具有高度敏感性的意义。 特别是细胞色素 P450对C. capistratus以其为食 柳珊瑚,否则由鱼类威慑保护 化感物质 这一发现具有潜在的广泛意义 以了解海洋中的捕食和食草动物的模式 环境 C. capistratus P450和 开发分子探针来测试 因此,亚致死水平的化学品适用于重要的 环境和生态问题。

项目成果

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