Pipalia Tapan G, Sultan Sami H A, Koth Jana, Knight Robert D, Hughes Simon M
Randall Centre for Cell & Molecular Biophysics, King's College London, London, UK.
Centre for Craniofacial and Regenerative Biology, King's College London, London, UK.
Methods Mol Biol. 2023;2640:227-248. doi: 10.1007/978-1-0716-3036-5_17.
Muscle regeneration models have revealed mechanisms of inflammation, wound clearance, and stem cell-directed repair of damage, thereby informing therapy. Whereas studies of muscle repair are most advanced in rodents, the zebrafish is emerging as an additional model organism with genetic and optical advantages. Various muscle wounding protocols (both chemical and physical) have been published. Here we describe simple, cheap, precise, adaptable, and effective wounding protocols and analysis methods for two stages of a larval zebrafish skeletal muscle regeneration model. We show examples of how muscle damage, ingression of muscle stem cells, immune cells, and regeneration of fibers can be monitored over an extended timecourse in individual larvae. Such analyses have the potential to greatly enhance understanding, by reducing the need to average regeneration responses across individuals subjected to an unavoidably variable wound stimulus.
肌肉再生模型揭示了炎症、伤口清除以及干细胞定向修复损伤的机制，从而为治疗提供了依据。虽然肌肉修复的研究在啮齿动物中最为先进，但斑马鱼正成为一种具有遗传和光学优势的额外模式生物。已经发表了各种肌肉损伤方案（化学和物理的）。在这里，我们描述了用于幼虫斑马鱼骨骼肌再生模型两个阶段的简单、廉价、精确、可适应且有效的损伤方案和分析方法。我们展示了在个体幼虫的较长时间过程中如何监测肌肉损伤、肌肉干细胞的侵入、免疫细胞以及纤维再生的示例。通过减少对受到不可避免的可变伤口刺激的个体的再生反应进行平均的需求，此类分析有可能极大地增进我们的理解。
