Development and validation of a fathead minnow liver cell line for environmental testing

林业、纸浆和造纸制造以及城市垃圾处理设施等第一产业雇用了大量加拿大人。它们的活动使用大量的水，当它们将水返回环境时，水中可能含有影响动物成功繁殖能力的化学物质。愚头鱼是用于预测水道中其他动物健康状况的实验中使用的模式物种。海洋行业和政府监管机构使用活的小鱼来筛选水中流出物的毒性。然而，这些测试既昂贵又繁琐。脊椎动物中参与环境污染物解毒的主要器官是肝脏，它对低水平的这些化合物非常敏感。已知毒素应答基因被激活或抑制，以响应毒物的攻击，但其中许多基因的特性尚不清楚。我们目前正在对小鱼的肝脏进行鉴定，使用一种名为DNA微阵列的技术来检查基因表达。为了简化和降低活体检测的成本，我们建议开发一种愚蠢的迷你肝细胞培养。这些细胞将在实验室中培养，很可能具有与鱼类肝脏细胞相同的毒物反应特征。然而，它们将更容易种植和用于常规毒理学测试。一些初步工作建议采用细胞系进行环境监测，但该项目将是第一个比较细胞和完整组织中基因表达的项目之一。这是采用这项技术之前的必要前提。我们的目标是验证这些细胞作为FISH检测的替代品的使用。这项测试的开发将降低加拿大工业环境监测的成本，从而有助于保护对加拿大经济至关重要的行业的就业。采用这种测试还将减少用于毒理学测试的实验室动物的数量，这是世界各地监管机构的共同目标。

Primary industries such as forestry, pulp and paper manufacturing and municipal waste treatment facilities employ significant numbers of Canadians. Their operations use large amounts of water and when they return it to the environment it may contain chemicals that affect the ability of animals to successfully reproduce. The fathead minnow is a model species used in experiments to predict the health of other animals in waterways.  Industry and government regulatory agencies use live minnows to screen aquatic effluents for toxicity.  However, these tests are expensive and cumbersome. The primary organ in vertebrates involved in detoxification of environmental contaminants is the liver, and it is very sensitive to low levels of thesecompounds. It is known that toxin-responsive genes are activated or repressed in response to challenge by toxicants, but the identity of many of these genes is unknown. We are currently performing this identification in minnow livers, using a technique to examine gene expression known as DNA microarrays. To simplify and lessen the cost of live testing, we propose to develop a fathead minnow liver cell culture. These cells will be grown in the laboratory and will likely possess the same toxicant-responsive characteristics as cells from livers in fish. However, they will be much easier to grow and use for routine toxicology testing. Some preliminary work has suggested the adoption of cell lines for environmental monitoring, but this project will be among the first to compare gene expression in cell versus intact tissue. This is a necessary prerequisite prior to adopting this technology. Our goal is to validate the use of these cells as a replacement for testing using fish. Thedevelopment of this test will lower the cost of environmental monitoring for Canadian industry, thus helping to safeguard jobs in sectors of critical economic importance to the Canadian economy. Adoption of this test will also reduce the number of lab animals used for toxicology testing, which is a common goal of regulatory agencies worldwide.