Developing and Using Sheepshead Minnow Microarrays for Ecotoxicology

描述（由申请人提供）：微阵列是测量环境中污染物影响的有力方法，羊头鲦鱼（Cyprinodon variegatus）是盐水环境测试中最常用的物种。在本提案中，我们解释了如何在羊头鲦鱼中开发和验证大型微阵列（5000 +基因）。我们将使用几种人为化合物（芘、铜、镉和双酚a）对羊头鲦鱼进行短期和长期暴露，然后使用微阵列测量这些化合物的基因表达特征。这些数据，连同各种生理端点，将成为关系数据库的基础。我们将分析数据并破译各种暴露产生的模式，以识别每种化合物的独特指纹。微阵列在羊头鲦鱼中的应用将使EcoArray公司为环境重要的水生物种提供的微阵列产品更加完善。环境污染物的测量和分析对美国环保署的超级基金监测活动非常重要，而羊头鲦鱼是沿海超级基金监测中经常使用的重要物种。此外，在羊头鲦鱼等前哨物种的基因水平上诊断生态毒理学效应，为未来将生态毒理学与人类健康联系起来提供了希望，这是国家毒理学基因组学中心的一个目标。我们的研究小组和其他人的实验表明，微阵列可以用来检测暴露于污染物引起的基因表达变化，很明显，污染物具有独特的遗传特征。由于许多污染物在基因水平上通过受体介导和非受体介导的途径诱导或抑制基因表达，微阵列可以帮助阐明受影响的信号通路。总的来说，微阵列提供了一种直接、有效的方法来提供有关环境对动物的生物效应的详细数据。此外，使用微阵列的分析通常比目前的测试方法便宜得多。随着这笔拨款的成功完成，我们将把羊头鲦鱼微阵列及其数据库纳入我们现有的产品线，并将其出售给EPA， USGS，学术界的研究人员以及对化合物筛选和环境监测/修复感兴趣的工业企业。

DESCRIPTION (provided by applicant): Microarrays are a powerful way to measure the impact of contaminants in the environment, and sheepshead minnows (Cyprinodon variegatus) are the most commonly used species in salt water environmental testing. In this proposal, we explain how we will develop and validate a large microarray (5,000+ genes) in sheepshead minnows. We will conduct a number of short and long term exposures on sheepshead minnows using several anthropogenic compounds (pyrene, copper, cadmium, and bisphenol A), then use the microarrays to measure the gene expression signatures for these compounds. This data, along with a variety of physiological endpoints, will be the basis for a relational database. We will analyze the data and decipher the patterns resulting from the various exposures to identify the unique fingerprints for each compound. Microarrays in sheepshead minnows will round out EcoArray's offering of microarrays for environmentally significant aquatic species. Measurement and analysis of environmental contaminants is very important to the EPA in its Superfund monitoring activities, and the sheepshead minnow is an important species that is routinely used for the monitoring of coastal superfund sites. In addition, diagnosis of ecotoxological effects at the gene level in sentinel species like sheepshead minnow offer the promise of future ability to tie ecotoxicology to human health, a goal of the National Center for Toxicogenomics. Experiments by our research group and others have shown that microarrays can be used to detect changes in gene expression caused by exposure to contaminants, and it is clear that contaminants have unique genetic signatures. Because many contaminants act at the gene level to induce or repress gene expression through both receptor-mediated and non-receptor mediated pathways, microarrays can help to elucidate signaling pathways that are affected. In general, microarrays offer a direct, effective way to provide detailed data about the biological effects of the environment on animals. In addition, analysis using microarrays is generally considerably less expensive than current testing methods. With the successful completion of this grant, we will incorporate the sheepshead minnow microarray and its database into our existing product line and sell it to the EPA, USGS, researchers in academia, as well as industrial concerns interested in compound screening and environmental monitoring/remediation. This project will result in a fully developed microarray for ecotoxicology testing in salt water. This microarray, together with the database this project will begin, can provide detailed, gene-level data on the biological impact of a chemical or an environment. In so doing, it can lead to assessment not only of water quality, but also of the implications of chemicals and environments for human health.