Abstract：
There is global demand for novel ecotoxicity testing tools that are based on alternative to animal models, have high throughput potential, and may be applicable to a wide diversity of taxa. Here we scaled up a microplate-based cell-free neurochemical testing platform to screen 800 putative endocrine disrupting chemicals from the U.S. Environmental Protection Agency\'s ToxCast e1k library against the glutamate (NMDA), muscarinic acetylcholine (mACh), and dopamine (D2) receptors. Each assay was tested in cellular membranes isolated from brain tissues from a representative bird (zebra finch = Taeniopygia castanotis), mammal (mink = Neogale vison), and fish (rainbow trout = Oncorhynchus mykiss). The primary objective of this short communication was to make the results database accessible, while also summarising key attributes of assay performance and presenting some initial observations. In total, 7200 species-chemical-assay combinations were tested, of which 453 combinations were classified as a hit (radioligand binding changed by at least 3 standard deviations). There were some differences across species, and most hits were found for the D2 and NMDA receptors. The most active chemical was C.I. Solvent Yellow 14 followed by Diphenhydramine hydrochloride, Gentian Violet, SR271425, and Zamifenacin. Nine chemicals were tested across multiple plates with a mean relative standard deviation of the specific radioligand binding data being 24.6%. The results demonstrate that cell-free assays may serve as screening tools for large chemical libraries especially for ecological species not easily studied using traditional methods.
摘要：
全球需要基于替代动物模型的新型生态毒性测试工具，具有高吞吐量潜力，并且可能适用于广泛的分类单元。在这里，我们扩大了基于微孔板的无细胞神经化学测试平台，从美国环境保护署的ToxCaste1k文库中筛选了800种推定的内分泌干扰化学物质，以对抗谷氨酸（NMDA）。毒蕈碱乙酰胆碱（mACh），和多巴胺（D2）受体。在从代表性鸟类（斑马雀=Taeniopygiacastanotis）的脑组织中分离的细胞膜中测试了每种测定，哺乳动物（水貂=Neogalevison），和鱼（虹鳟鱼=Oncorhynchusmykiss）。这种简短交流的主要目的是使结果数据库易于访问，同时还总结了测定性能的关键属性并提出了一些初步观察结果。总的来说,测试了7200个物种-化学分析组合，其中453个组合被分类为命中（放射性配体结合变化至少3个标准偏差）。不同物种之间存在一些差异，发现了D2和NMDA受体的大多数命中。最活跃的化学物质是C.I.溶剂黄14，其次是盐酸苯海拉明，龙胆紫罗兰,SR271425和扎米那新。在多个平板上测试9种化学品,特异性放射性配体结合数据的平均相对标准偏差为24.6%。结果表明，无细胞测定可以作为大型化学文库的筛选工具，尤其是对于使用传统方法不易研究的生态物种。
