NTP MediumThroughput C. elegans Screening Facility

目前，有超过80，000种化学品在使用，每年大约有2000种新化学品投入使用。 监管机构已经认识到需要替代毒理学方法和模型，以减少当前毒性测试方案的时间和费用。 在国家毒理学计划（NTP）的合作下，我们的小组正在开发C。秀丽隐杆线虫作为体内毒理学测试的替代生物。短的生命周期，容易和廉价的维护和培养，以及详细的生物学知识，允许开发快速，低成本的毒性测试，很容易借给自己的机制研究毒物的行动。由于应激反应和其他相关途径在进化上的保守性，C. elegans将适用于理解包括人类在内的高等生物的类似过程。 该小组是国家毒理学计划的一部分，并在新的生物分子筛选分支。 筛选设施参与了C。线虫作为毒理学测试的替代生物。 由于信号转导和应激反应途径在进化上的保守性，很可能C。elegans将适用于理解高等生物中的类似过程。 该小组的主要活动有两个：研究和开发C。elegans MTS和HTS技术以及参与NTP、EPA和NCGC之间的谅解备忘录，指定为Tox 21（Science 15 February 2008 319：906-907 DOI：10.1126/science.1154619）。 1. C.线虫中等通量筛选试验--已经开发了用于监测生长、大小、繁殖、进食和运动的方案，并已用于测试100种毒物。 在监测方案的制定中，专门为C。elegans数据。 我们已提交了一份关于摄食试验的手稿，并正在完成其他研究，这些研究将纳入目前正在编写的关于生殖和生长试验的手稿中。 在与Chris Portier（LMT），Grace Kissling（EDBP），Marjolein Smith（ASI）的合作中，我们开发了一个描述C。线虫的生长和毒物对生长参数的影响。 我们目前正在产生一个收集的绿色荧光蛋白为基础的，应激反应转基因C。优雅线条 这些C线。线虫将用于鉴定毒物调节已知影响生物体发育的信号传导和应激反应途径的活性的能力（例如，MAPK、p53、酪氨酸磷酸酶）。 2. 国家毒理学计划HTS教师-我参加了NTP/EPA/NCGC谅解备忘录。 我们已经开始开发使用C的协议。elegans作为HTS测试生物。这包括开发新的取样格式、统计工具和质量保证/质量控制协议。我们目前正在测试NTP 1408和ToxCast 320农药活性物质化学套件。

Currently, there are over 80,000 chemicals in use and approximately 2000 new chemicals are introduced into use every year. Regulatory agencies have recognized the need for alternative toxicological methods and models to decrease the time and expense of current toxicity testing protocols. In association with the National Toxicology Program (NTP), our group is developing C. elegans as an alternative organism for in vivo toxicological testing. Short life cycles, easy and inexpensive maintenance and culturing, and detailed biological knowledge has allowed for the development of rapid, low-cost toxicity tests that readily lend themselves to mechanistic studies of toxicant actions. Because of the evolutionarily conserved nature of the stress-response and other relevant pathways, it is likely that responses elicited in C. elegans will be applicable to understanding similar processes in higher organisms, including humans. This group is part of the National Toxicology Program and within the new Biomolecular Screening Branch. The screening facility is involved in the development of C. elegans as an alternative organism for toxicological testing. Because of the evolutionarily conserved nature of signal transduction and stress-response pathways, it is likely that responses elicited in C. elegans will be applicable to understanding similar processes in higher organisms. There are two major activities of this group: research and development of C. elegans MTS and HTS technologies and participation in the MOU among the NTP, EPA, and NCGC, designated Tox21 (Science 15 February 2008 319: 906-907 DOI: 10.1126/science.1154619). 1. Development of C. elegans medium-throughput screening assays - Protocols for the monitoring of growth, size, reproduction, feeding, and movement have been developed and have been used to test 100 toxicants. Included in the creation of monitoring protocols, statistical analysis routines have been developed specifically for the C. elegans data. We have submitted one manuscript on the feeding assay and are completing additional studies that will be included in manuscripts currently being prepared on the reproduction and growth assays. In collaboration with Chris Portier (LMT), Grace Kissling (EDBP), Marjolein Smith (ASI) we have developed a mathematical model that described C. elegans growth and the effects of toxicants on growth parameters. We are currently generating a collection of green fluorescent protein-based, stress-responsive transgenic C. elegans lines. These lines of C. elegans will be used to identify the ability of toxicants to modulate the activities of signaling and stress-responsive pathways know the affect organism development (e.g., MAPK, p53, tyrosine phosphatase). 2. National Toxicology Program HTS faculty - I am participating in the NTP/EPA/NCGC MOU. We have begun developing protocols to use C. elegans as an HTS test organism. This includes the development of new sampling formats, statistical tools and QA/QC protocols. We are currently testing the NTP 1408 and the ToxCast 320 pesticide actives chemical sets.