Finding effective analgesics in zebrafish by analysing effects on the nervous system (CNS)

斑马鱼在生物医学研究中正变得越来越受欢迎(根据内政部的数据，2011年有超过400.000只斑马鱼被使用)，因为它相对较小，而且仍然与其他脊椎动物(脊椎动物)有许多相似之处。这使得斑马鱼的科学发现可以翻译成医学研究。成鱼和幼鱼被用于不同的领域，包括癌症和神经退行性疾病的研究，并经历了潜在的痛苦程序，但大多数程序都没有适当的止痛(止痛)。这主要是因为人们对鱼类如何处理潜在的有害物质知之甚少。例如，正在努力研究潜在痛苦经历后行为的变化以及特定药物(止痛剂)如何缓解这些变化，以便制定对鱼类有效的止痛(止痛)方案。然而，接受这些刺激、处理它们并指导行为变化的中枢神经系统并没有得到分析。我们认为，分析中枢神经系统的变化对于直接证明有害刺激影响大脑是必不可少的，而且止痛药在大脑水平上是有效的。在这个试点项目中，我们的目标是通过发现潜在痛苦经历后不同基因在脊髓和大脑区域的活性变化来支持分析行为和其他参数的努力。这将提供有关处理此类输入的直接信息。此外，通过尝试一些潜在的止痛药，我们将通过直接测量大脑中的基因活动来确定其中哪种最有效。我们的初步分析表明，将芥子油应用于肌肉组织会增加脊髓和脑干中基因的活性，这些基因通常在有害刺激的背景下被激活。理想情况下，我们会发现止痛剂已经在脊髓水平上阻断了这种反应，刺激首先在脊髓水平上进行处理。在随后的研究中，我们将把来自大脑的发现与行为和其他非侵入性措施的数据进行比较，以确定鱼类止痛的最佳方案，并在未来的斑马鱼研究中得出成功止痛的非侵入性指标。因此，我们的数据将有助于在3R中定义的精细化意义上改进鱼类实验，通过设计有神经生物学观察支持的止痛方案，从而向科学家和政策制定者通报鱼类实验的最佳实践。

The zebrafish is becoming increasingly popular in biomedical research (more than 400.000 individuals were used in 2011 according to the Home Office), as it is relatively small and still shares many features with other back-boned animals (vertebrates). This makes scientific findings from the zebrafish translatable for medical research. Adult and larval fish are being used in diverse fields, including research in cancer and neurodegenerative diseases and undergo potentially painful procedures, yet most procedures are done without proper analgesia (pain relief). This is mainly because not much is known about how fish process potentially harmful input. Efforts are underway to look, for example, at changes in behaviour after potentially painful experiences and how specific drugs (analgesics) mitigate these, in order to develop protocols for effective analgesia (pain relief) in fish. However, the central nervous system, which receives these stimuli, processes them and directs the behavioural changes, is not being analysed. We believe that analysing changes in the central nervous system is indispensable to directly demonstrate that harmful stimuli affect the brain, and that analgesic drugs are effective at the level of brain. In this pilot project, we aim to support efforts analysing behaviour and other parameters by finding changes in the activity of different genes in areas of the spinal cord and brain after potentially painful experiences. This will give direct information on the processing of such input. Moreover, by trying a number of potential analgesics we will determine which of these work most effectively by directly measuring gene activity in the brain. Our preliminary analyses show that application of mustard oil to muscle tissue leads to increased activity of genes in the spinal cord and brainstem that are often activated in the context of harmful stimuli. Ideally, we will find analgesics that block this reaction already at the level of the spinal cord, where stimuli are first processed. In subsequent research, our findings from the brain will be compared to data from behaviour and other non-invasive measures to determine optimal protocols for analgesia in fishes and derive non-invasive indicators of successful analgesia in future zebrafish research. Our data will thus help to refine experiments in fishes in the sense of Refinement as defined in the 3Rs, by devising protocols for analgesia that are supported by neurobiological observations and thus inform scientists and policy makers on best practice in fish experiments.