Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, 185 Meeting Street, Providence, RI, 02912, USA.
Center for Computation and Visualization, Brown University, Providence, RI, USA.
Sci Rep. 2023 Feb 23;13(1):3174. doi: 10.1038/s41598-023-30303-w.
Brain function studies greatly depend on quantification and analysis of behavior. While behavior can be imaged efficiently, the quantification of specific aspects of behavior is labor-intensive and may introduce individual biases. Recent advances in deep learning and artificial intelligence-based tools have made it possible to precisely track individual features of freely moving animals in diverse environments without any markers. In the current study, we developed Zebrafish Larvae Position Tracker (Z-LaP Tracker), a modification of the markerless position estimation software DeepLabCut, to quantify zebrafish larval behavior in a high-throughput 384-well setting. We utilized the high-contrast features of our model animal, zebrafish larvae, including the eyes and the yolk for our behavioral analysis. Using this experimental setup, we quantified relevant behaviors with similar accuracy to the analysis performed by humans. The changes in behavior were organized in behavioral profiles, which were examined by K-means and hierarchical cluster analysis. Calcineurin inhibitors exhibited a distinct behavioral profile characterized by increased activity, acoustic hyperexcitability, reduced visually guided behaviors, and reduced habituation to acoustic stimuli. The developed methodologies were used to identify 'CsA-type' drugs that might be promising candidates for the prevention and treatment of neurological disorders.
大脑功能研究在很大程度上依赖于行为的量化和分析。虽然行为可以被有效地成像，但行为的特定方面的量化是劳动密集型的，并且可能引入个体偏差。最近在深度学习和基于人工智能的工具方面的进展使得有可能在没有任何标记的情况下精确地跟踪不同环境中自由移动的动物的个体特征。在当前的研究中，我们开发了 Zebrafish Larvae Position Tracker (Z-LaP Tracker)，这是对无标记位置估计软件 DeepLabCut 的修改，用于在高通量 384 孔设置中量化斑马鱼幼虫的行为。我们利用了我们的模型动物，斑马鱼幼虫的高对比度特征，包括眼睛和卵黄，用于我们的行为分析。使用这种实验设置，我们以类似于人类分析的准确性量化了相关行为。行为变化被组织成行为特征，通过 K-means 和层次聚类分析进行检查。钙调神经磷酸酶抑制剂表现出一种独特的行为特征，表现为活动增加、听觉超兴奋性、视觉引导行为减少以及对听觉刺激的习惯化减少。开发的方法学用于鉴定“CsA 型”药物，这些药物可能是预防和治疗神经障碍的有希望的候选药物。
