de Carvalho Ana Cláudia Costa, Morato Sílvio, Gouveia Amauri, Walsh-Monteiro André
Laboratory of Neurochemistry and Behaviour, Tucuruí Campus, Federal Institute of Pará, Tucuruí, Brazil.
Laboratory of Neuroscience and Behaviour, Center for Theory and Behaviour Research, Federal University of Pará, Belém, Brazil.
Zebrafish. 2025 Jun;22(3):86-98. doi: 10.1089/zeb.2025.0002. Epub 2025 May 15.
Over the last few decades, zebrafish have proven to be a valuable tool for biomedical research, with advantages mainly in the use of embryos and adults. The larval stage, on the contrary, is somewhat less used, generally due to a lack of protocols compared with the other stages. In this study, we propose a protocol to study anxiety-like behavior in larvae using the plus maze with ramp (PMR). In the PMR, anxious behavior is measured by the time spent on the flat arms relative to the ramped arms of the apparatus. In the first phase of the study, animals at 5, 14, and 21 days postfertilization (dpf) were exposed to the PMR at different water column heights and session times. It was observed that animals at 14 and 21 dpf were more sensitive to the PMR and exhibited obvious anxiety-like behavior compared with larvae at 5 dpf. In the second phase of the study, to assess the robustness of the type-anxious response exhibited by the larvae, animals at 5, 10, and 14 dpf treated with anxiolytic drugs were exposed to PMR. Alcohol exposure showed a bimodal effect for animals at 10 and 14 dpf, with an anxiolytic effect at intermediate doses, with more exploration of the apparatus and time in the ramp arms. Higher doses caused a reduction in locomotor activity characteristic of drunkenness. Exposure to clonazepam produced anxiolytic effects and a reduction in locomotor activity at the highest dose in 10 and 14 dpf animals. On the contrary, 5 dpf animals showed unexpected effects, which we believe to be related to their stage of development. The results indicate that the PMR is an effective tool for assessing anxiety-like behavior in zebrafish larvae and is sensitive to anxiolytic drugs. Furthermore, anxiety-like behavior appears to vary during the early stages of larval development.
在过去几十年中，斑马鱼已被证明是生物医学研究的一种有价值的工具，其优势主要体现在胚胎和成年斑马鱼的使用上。相反，幼体阶段的使用相对较少，这通常是由于与其他阶段相比缺乏相关实验方案。在本研究中，我们提出了一种使用带斜坡的十字迷宫（PMR）来研究幼体焦虑样行为的实验方案。在PMR中，焦虑行为通过动物在装置的平臂上相对于斜坡臂上所花费的时间来衡量。在研究的第一阶段，将受精后5天、14天和21天（dpf）的动物置于不同水柱高度和实验时间的PMR中。观察到14 dpf和21 dpf的动物对PMR更敏感，与5 dpf的幼体相比表现出明显的焦虑样行为。在研究的第二阶段，为了评估幼体表现出的焦虑样反应的稳健性，将用抗焦虑药物处理的5 dpf、10 dpf和14 dpf的动物置于PMR中。酒精暴露对10 dpf和14 dpf的动物呈现出双峰效应，中等剂量时有抗焦虑作用，表现为对装置的探索增加以及在斜坡臂上的时间增加。高剂量则导致醉酒特征的运动活动减少。给予氯硝西泮对10 dpf和14 dpf的动物在最高剂量时产生抗焦虑作用并降低运动活动。相反，5 dpf的动物表现出意想不到的效果，我们认为这与它们的发育阶段有关。结果表明，PMR是评估斑马鱼幼体焦虑样行为的有效工具，并且对抗焦虑药物敏感。此外，焦虑样行为在幼体发育的早期阶段似乎有所不同。
