The current study developed and evaluated a conditioning protocol for zebrafish which can be used in groups of fish instead of isolated individuals. The protocol consisted of the application of a neutral conditioned stimulus (CS), i.e. a novel object in the tank, followed by an aversive, stressful unconditioned stimulus (US), i.e. chasing, in groups of fish in experimental tanks. This training protocol was applied to fish once per day for one- or three- consecutive days. The following day fish were tested for the display of behavioural, i.e. bottom dwelling, and neuroendocrine, i.e. cortisol and stress-related gene expression, conditioned responses. Results showed that one-day of conditioning was not adequate to produce conditioned responses. Repetition of training sessions for three days, however, led to the display of conditioned responses upon exposure to the conditioning stimulus alone. Specifically, when exposed to the CS, paired fish showed increased bottom dwelling, as well as elevated trunk cortisol concentration and c-fos gene expression in the brain. Data suggest that the applied CS can be perceived by fish as a signal to prepare anticipatory behavioural and neuroendocrine responses to cope with the expected aversive event. Finally, this study is one of the few to test associative learning in groups of zebrafish, thus avoiding training and testing fish individually which can be stressful and impede normal behaviour.

中文翻译：

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成群结队的成年斑马鱼 Danio rerio 对调节方案的行为和生理反应

目前的研究开发并评估了斑马鱼的调理方案，该方案可用于鱼群而不是孤立的个体。该协议包括应用中性条件刺激 (CS)，即坦克中的新物体，然后是厌恶的、有压力的无条件刺激 (US)，即在实验坦克中追逐鱼群。该训练方案每天一次应用于鱼，连续一天或三天。第二天对鱼的行为表现（即底部栖息）和神经内分泌（即皮质醇和压力相关基因表达）条件反应的表现进行测试。结果表明，一天的条件反射不足以产生条件反应。然而，重复三天的培训课程，导致在单独暴露于条件刺激时显示条件反应。具体来说，当暴露于 CS 时，成对的鱼表现出增加的底部栖息地，以及大脑中躯干皮质醇浓度和 c-fos 基因表达的升高。数据表明，应用的 CS 可以被鱼感知为准备预期行为和神经内分泌反应的信号，以应对预期的厌恶事件。最后，这项研究是少数在斑马鱼群体中测试联想学习的研究之一，从而避免单独训练和测试鱼，这可能会带来压力并阻碍正常行为。数据表明，应用的 CS 可以被鱼感知为准备预期行为和神经内分泌反应的信号，以应对预期的厌恶事件。最后，这项研究是少数在斑马鱼群体中测试联想学习的研究之一，从而避免单独训练和测试鱼，这可能会带来压力并阻碍正常行为。数据表明，应用的 CS 可以被鱼感知为准备预期行为和神经内分泌反应的信号，以应对预期的厌恶事件。最后，这项研究是少数在斑马鱼群体中测试联想学习的研究之一，从而避免单独训练和测试鱼，这可能会带来压力并阻碍正常行为。