Rapid Zebrafish Behavioral Profiling Assay Accelerates the Identification of Environmental Neurodevelopmental Toxicants
Rapid Zebrafish Behavioral Profiling Assay Accelerates the Identification of Environmental Neurodevelopmental Toxicants
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DOI：
10.1021/acs.est.0c06949
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发表时间：
2021-01-20
影响因子：
11.4
通讯作者：
Werdich, Andreas A.
中科院分区：
文献类型：
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作者：
Zhang, Kun;Liang, Jiahui;Werdich, Andreas A.
筛选潜在环境神经发育毒物的快速且具有成本效益的体内测定对于解决体外平台的局限性是必要的，例如无法完全重现整个生物体的发育和生理过程。本研究建立了一种快速的斑马鱼行为分析方法，通过定量评价斑马鱼胚胎的多种自发运动特征来表征环境物质对斑马鱼神经发育的影响。这种基于视频分析的检测方法自动分割每个胚胎，因此能够准确量化自发运动特征，包括频率，持续时间，强度，间隔和连续运动的数量。当用八种已知是神经发育毒物的环境物质（如毒死蜱和双酚A）进行测试时，该检测成功地捕获了在以前的研究中有充分记录的频率变化，同时还提供了额外的信息。使用优化的程序，我们进一步评估了132种潜在的神经毒素，这些神经毒素跨越了广泛的分子靶点，其中许多是以前在环境水体中检测到的。不同的行为条形码表明，自发运动受到不同的神经活性物质的影响，并且可以用开发的测定法有效地评估这些影响。基于网络的工具，名为EMAnalysis，进一步提供在http://www.envh.sjtu.edu.cn/zebrafish_contraction.jsp。因此，该测定提供了一个有效的平台，以加快神经毒性环境污染物的发现。
Rapid and cost-effective in vivo assays to screen potential environmental neurodevelopmental toxicants are necessary to address the limitations of in vitro platforms, such as the inability to fully recapitulate the developmental and physiological processes of whole organisms. In the present study, a rapid zebrafish behavioral profiling assay was developed to characterize the neurodevelopmental effects of environmental substances by quantitatively evaluating multiple spontaneous movement features of zebrafish embryos. This video analysis-based assay automatically segmented every embryo and thus was able to accurately quantify spontaneous movement features, including frequency, duration, intensity, interval, and the number of continuous movements. When tested with eight environmental substances known to be neurodevelopmental toxicants, such as chlorpyrifos and bisphenol A, the assay successfully captured frequency alterations that were well-documented in previous studies while also providing additional information. Using an optimized procedure, we further assessed 132 potential neurotoxins that spanned a wide range of molecular targets, many of which were previously detected in environmental waterbodies. The distinct altered behavioral barcodes indicated that the spontaneous movement was impacted by diverse neuroactive substances, and the effects could be effectively evaluated with the developed assay. The web-based tool, named EMAnalysis, is further provided at http://www.envh.sjtu.edu.cn/zebrafish_contraction.jsp. Thus, this assay provides an efficient platform to accelerate the pace of neurotoxic environmental contaminant discoveries.