Developing and Using Sheepshead Minnow Microarrays for Ecotoxicology

描述(申请人提供)：微阵列是测量环境中污染物影响的有效方法，而羊头小鱼(Cyprinodon Varegatus)是咸水环境测试中最常用的物种。在这个提案中，我们解释了我们将如何开发和验证羊头小鱼的大型微阵列(5,000多个基因)。我们将使用几种人为化合物(芘、铜、镉和双酚A)对羊头小鱼进行短期和长期暴露，然后使用微阵列来测量这些化合物的基因表达特征。这些数据以及各种生理终点将成为关系数据库的基础。我们将分析数据并破译各种曝光产生的模式，以确定每种化合物的唯一指纹。羊头小鱼中的微阵列将使EcoArray为对环境具有重要意义的水生物种提供的微阵列更加完善。环境污染物的测量和分析对环保局的超级基金监测活动非常重要，而羊头小鱼是一种重要的物种，经常用于沿海超级基金地点的监测。此外，在基因水平上对绵羊头鱼等前哨物种的生态毒理影响进行诊断，为未来将生态毒理学与人类健康联系在一起的能力提供了希望，这是国家毒物基因组学中心的目标。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在污染物中导致的基因表达变化，而且很明显，污染物具有独特的遗传特征。由于许多污染物通过受体介导和非受体介导的途径在基因水平上诱导或抑制基因表达，因此微阵列可以帮助阐明受影响的信号通路。一般来说，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影响的详细数据。此外，使用微阵列进行分析通常比目前的检测方法便宜得多。随着这笔赠款的成功完成，我们将把羊头迷你微阵列及其数据库整合到我们现有的产品线中，并将其出售给美国环保局、美国地质调查局、学术界的研究人员以及对化合物筛选和环境监测/补救感兴趣的行业公司。 该项目将产生一种完全开发的微阵列，用于在盐水中进行生态毒理学测试。这个微阵列，连同这个项目将开始的数据库，可以提供关于化学物质或环境的生物影响的详细的、基因水平的数据。在这样做的过程中，不仅可以评估水质，而且还可以评估化学品和环境对人类健康的影响。

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.