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Author:
Date:
2017-04-05
[Abstract] The objective of this protocol is to provide a detailed description for the construction and use of a behavioral apparatus, the zBox, for high-throughput behavioral measurements in larval zebrafish (Danio rerio). The zBox is used to measure behavior in multiple individuals simultaneously. Individual fish are housed in wells of multi-well plates and receive acoustic/vibration stimuli with simultaneous recording of behavior. Automated analysis of behavioral movies is performed with MATLAB scripts. This protocol was adapted from two of our previously published papers (Levitz et al., 2013; Pantoja et al., 2016). The zBox provides an easy to setup flexible platform for behavioral experiments in zebrafish larvae.
[摘要] 该协议的目的是提供一个行为装置的构造和使用的详细描述，zBox，用于幼虫斑马鱼（Danio rerio）中的高通量行为测量。 zBox用于同时测量多个人的行为。 个体鱼被放置在多孔板的孔中并且接收声/振动刺激同时记录行为。 使用MATLAB脚本对行为电影进行自动分析。 这个协议是从我们之前发表的两篇文章（Levitz et al。，2013; Pantoja et al。，2016）改编而成。 zBox为斑马鱼幼虫的行为实验提供了一个易于设置的灵活平台。
【背景】人口中的个体之间的行为差异是普遍存在的，并且可能在适应进化过程中不同选择压力时发挥作用。 然而，当群体的行为的定量被表示为手段和相关的分散体时，往往忽略个体之间行为的变化。 在这个协议中，我们描述了一种在斑马鱼（Danio rerio）幼虫的群体级别的声学惊吓行为习惯中表征个性的方法。 我们的方法和设置可以轻松适应其他斑马鱼幼虫行为的个性研究。
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Author:
Date:
2016-10-05
[Abstract] The production of free radicals is the result of normal cellular metabolism. Free radicals are involved in innumerable different cellular and biological functions such as signaling, proliferation, cell death, aging, inflammation, etc. Under physiological conditions, the levels of reactive oxygen species (ROS) are strictly regulated by the cells. However, during stressful conditions such as oxidative stress, ROS levels increase causing damages to different molecules like DNA, lipids and proteins. Increased levels of ROS have been associated with a growing list of different diseases. In this protocol, we used MitoSOX and CellROX Green oxidative stress probes to label the intracellular ROS and detect the fluorescence using cell sorting and confocal analyses.
[摘要] 自由基的产生是正常细胞代谢的结果。 自由基涉及无数不同的细胞和生物学功能，例如信号传导，增殖，细胞死亡，衰老，炎症等。 在生理条件下，活性氧（ROS）的水平被细胞严格调节。 然而，在应激条件例如氧化应激期间，ROS水平增加导致对不同分子如DNA，脂质和蛋白质的损伤。 增加的ROS水平与不断增加的不同疾病列表相关联。 在这个协议中，我们使用MitoSOX和CellROX绿色氧化应激探针标记细胞内ROS和使用细胞分选和共聚焦分析检测荧光。
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