Name: Analysis of Recorded Videos for Behavioral Testing of Adult Zebrafish
Uploaded: 2023-11-03T12:40:00
Description: Watch this Scientific Journal Video about Analysis of Recorded Videos for Behavioral Testing of Adult Zebrafish at JoVE.com

analysis of recorded videos for behavioral testing of adult zebrafish

行为的：评估成年斑马鱼神经毒性作用的测试电池

The zebrafish (Danio rerio) is a popular model vertebrate species for ecotoxicology, drug discovery, and safety pharmacology studies. Its low cost, well-established molecular genetic tools, and conservation of key physiological processes involved in the morphogenesis and maintenance of the nervous system make zebrafish an ideal animal model for neuroscience research, including neurobehavioral toxicology1,2. The main endpoint for evaluating the neurotoxicity of a chemical was, until recently, the presence of neuropathological effects. Lately, however, behavioral endpoints have been incorporated into neurotoxicological screening protocols, and these functional outcomes are now commonly used to identify and determine the potential neurotoxicity of chemicals3,4. Moreover, behavioral endpoints are highly relevant from an ecological point of view, as even a very mild behavioral change in fish could endanger the survival of the animal in natural conditions5.
One of the most used behavioral assays in adult zebrafish research is the novel tank test (NTT), which measures anxiety-like behavior6,7. In this assay, fish are exposed to novelty (fish are placed in an unfamiliar tank), a mild aversive stimulus and their behavioral responses are observed. NTT is used to assess basal locomotor activity, geotaxis, freezing, and erratic movements of fish, principally. Erratic8 is characterized by abrupt changes of direction (zigzagging) and repeated episodes of accelerations (darting). It is an alarm reaction and is usually observed before or after freezing episodes. Freezing behavior corresponds to a complete cessation of the fish&#39;s movements (except for opercular and ocular movements) while on the bottom of the tank, as distinguished from immobility caused by sedation, which causes hypolocomotion, akinesia, and sinking8. Freezing is usually related to a high state of stress and anxiety and is also part of submissive behavior. Complex behaviors are excellent indicators of the state of anxiety of animals. NTT has been shown to be sensitive to pharmacological and genetic manipulation9, making it a valuable tool for studying the neural basis of anxiety and related disorders.
Zebrafish are a highly social species, so we can measure a wide range of social behaviors. The shoaling test (ST) and the social preference test (SPT) are the most used assays to assess social behavior10. The ST measures the tendency of fish to group together11 by quantifying their spatial behavior and movement patterns. ST is useful for studying group dynamics, leadership, social learning, and understanding the social behavior of many fish species12. The SPT in adult zebrafish was adapted from Crawley&#39;s preference for social novelty test for mice13&#160;and quickly became a popular behavioral assay for the study of social interaction in this model species14. These two tests have also been adapted for use in drug screening assays and have shown promise for identifying novel compounds that modulate social behavior15,16.
In general, behavioral assays in adult zebrafish are powerful tools that can provide valuable information on the behavior mechanisms or the neurophenotypes of active compounds and abused drugs17. This protocol details how to implement these behavioral tools7 with basic material resources and how to apply them in toxicity assays to characterize the effects of a wide range of neuroactive compounds. In addition, we will see that the same tests can be applied to assess the neurobehavioral effects of acute exposure to a neuroactive compound (methamphetamine) but also to characterize these effects after chronic exposure to environmental concentrations of a pesticide (glyphosate).

作者聚焦：成年斑马鱼环境和药物诱导的行为变化评估 

在单个水箱中使用一系列行为测试对成年 Danio rerio 进行神经毒性评估

Our main research aims to identify the behavioral effects induced by environmental pollutants and drugs in fish. Also trying to understand the mechanisms leading to these effects. A social and anxiety-like behavior cannot be conveniently addressed using embryos.We conduct studies targeting these behaviors in the adult Zebrafish model. This protocol details how to implement behavioral tools with basic material resources to facilitate the reproducibility of tests and harmonize the results obtained within the scientific community. The assays presented in this protocol have proven to be highly reproducible and sensitive to genetic and pharmacological manipulations, making them variable and noninvasive tools fostering the neural and molecular mechanisms underlying behavior.Once a behavioral effort has been identified, the most challenging part is to determine the specific modes of action leading to the observative efforts. Starting by the molecular initiating event where predictions can provide some clues on the molecular initiating event, we are still far from obtaining accurate predictions. We are currently interested in the determining the phase validity of the chemical models of Parkinson's disease developed in embryonic and adult Zebrafish by using MPTP and rotenone with the same, the kinematic of the gait-like movement or during an explosive movement like the acoustic evoked escape response, will be evaluated.

Watch this Scientific Journal Video about Analysis of Recorded Videos for Behavioral Testing of Adult Zebrafish at JoVE.com

Analysis of Recorded Videos for Behavioral Testing of Adult Zebrafish  

分析用于成年斑马鱼行为测试的录制视频

打开分析软件以检查鱼类行为测试数据。要详细说明新模板，请单击“从模板新建”，然后单击“应用预定义的模板”，然后单击“从视频文件”。然后从模板设置中选择一个视频。在参数窗口中，选择模型鱼到成年斑马鱼，将竞技场选择为开放场地，选择正方形到一个竞技场以及每个竞技场的受试者数量。然后按中心点选择检测类型，并将帧速率调整为 30 FPS。将实验命名为模板，并将其存储在与其余视频相同的文件夹中。在“实验设置”下，查看已配置的设置。在竞技场设置中，右键单击屏幕中央，然后选择抓取。从显示屏中的文件中，选择一个高质量的视频图像，然后单击“接受”以捕获此图像以进行背景设置。使用水箱的 19 厘米宽度作为刻度，校准捕获的图像以创建校准规则。然后绘制竞技场，并使用框架功能描绘形状区域。对于新型水箱试验和浅滩试验，在水箱上画两个相等的水平箱。为社会偏好测试绘制三个相等的垂直框。在“试验控制设置”下，放置无限时间条件并删除其他组件。接下来，在“检测设置”下，从视频文件中选择要分析的视频并检查检测质量。单击“自动检测”以调整检测并重新聚焦动物。在“试用设置”下，输入一个试用。然后在“数据配置文件”下，创建结果对话框窗口。在“分析配置文件”下，选择要分析的参数。要复制和使用模板，请使用行为分析软件打开模板文件。然后转到“文件”并单击“另存为”以创建一个新的相同文件。或者，选择“从模板新建”，然后应用于自定义模板。接下来，转到竞技场设置以重新调整模板。在“采集”下，选择“DDS”，然后单击“就绪”开始处理视频。采集完成后，转到跟踪编辑器以检查处理后的视频。最后，在“统计”中，单击“计算”，然后单击“导出数据”。在暴露于甲基苯丙胺的斑马鱼的新型水箱测试中评估的焦虑行为表现出行进距离、探索距离和时间以及访问水箱上部区域的次数减少。首次访问上层区域之前的潜伏时间显着增加。在接触甲基苯丙胺三小时后，冷冻行为也显着增加。在甲基苯丙胺处理鱼的浅滩测试中评估的社会行为显示出社会孤立的行为表型，个体之间的平均距离和最远距离增加。在甲基苯丙胺处理鱼的社会偏好测试中评估的社会行为显示，在同种区域花费的时间和行进距离显着减少。