Three-Dimensional Neurophenotyping of Adult Zebrafish Behavior

The use of adult zebrafish (Danio rerio) in neurobehavioral research is rapidly expanding. The present large-scale study applied the newest video-tracking and data-mining technologies to further examine zebrafish anxiety-like phenotypes. Here, we generated temporal and spatial three-dimensional (3D) reconstructions of zebrafish locomotion, globally assessed behavioral profiles evoked by several anxiogenic and anxiolytic manipulations, mapped individual endpoints to 3D reconstructions, and performed cluster analysis to reconfirm behavioral correlates of high- and low-anxiety states. The application of 3D swim path reconstructions consolidates behavioral data (while increasing data density) and provides a novel way to examine and represent zebrafish behavior. It also enables rapid optimization of video tracking settings to improve quantification of automated parameters, and suggests that spatiotemporal organization of zebrafish swimming activity can be affected by various experimental manipulations in a manner predicted by their anxiolytic or anxiogenic nature. Our approach markedly enhances the power of zebrafish behavioral analyses, providing innovative framework for high-throughput 3D phenotyping of adult zebrafish behavior.成年斑马鱼（Danio rerio）在神经行为学研究中的应用正快速拓展。本项大规模研究采用最新的视频追踪与数据挖掘技术，进一步探究斑马鱼的类焦虑表型。本研究生成了斑马鱼运动的时空三维（3D）重建结果，全局评估了多种致焦虑与抗焦虑干预手段所诱导的行为谱，将单个行为终点映射至三维重建结果中，并通过聚类分析再次验证了高、低焦虑状态对应的行为关联特征。三维游动路径重建技术的应用不仅提升了数据密度，同时整合了行为学数据，为斑马鱼行为的观测与表征提供了全新途径；该技术还可快速优化视频追踪参数设置，以提升自动化量化指标的准确性，同时表明斑马鱼游泳活动的时空组织模式可被各类实验干预手段调控，且其调控规律与干预手段的抗焦虑或致焦虑特性相符。本研究方法显著增强了斑马鱼行为学分析的效能，为成年斑马鱼行为的高通量三维表型分析提供了创新性研究框架。