Department of Developmental Biology, Washington University School of Medicine in St. Louis, St. Louis, MO, USA.
Center of Regenerative Medicine, Washington University School of Medicine in St. Louis, St. Louis, MO, USA.
Methods Mol Biol. 2024;2707:215-222. doi: 10.1007/978-1-0716-3401-1_14.
Due to their renowned regenerative capacity, adult zebrafish are a premier vertebrate model to interrogate mechanisms of innate spinal cord regeneration. Following complete transection to their spinal cord, zebrafish extend glial and axonal bridges across severed tissue, regenerate neurons proximal to the lesion, and regain swim capacity within 8 weeks of injury. Here, we describe methods to perform complete spinal cord transections and to assess functional and cellular recovery during regeneration. For spinal cord injury, a complete transection is performed 4 mm caudal to the brainstem. Swim endurance is quantified as a central readout of functional spinal cord repair. For swim endurance, zebrafish are subjected to a constantly increasing water current velocity until exhaustion, and time at exhaustion is reported. To assess cellular regeneration, histological examination is performed to analyze the extents of glial and axonal bridging across the lesion.
由于其出色的再生能力,成年斑马鱼成为研究先天脊髓再生机制的首选脊椎动物模型。在对其脊髓进行完全横断后,斑马鱼会在切断的组织中延伸胶质和轴突桥,在损伤后的 8 周内再生受损部位附近的神经元,并恢复游泳能力。在这里,我们将描述进行完全脊髓横断的方法,并评估再生过程中的功能和细胞恢复情况。对于脊髓损伤,在脑干后 4 毫米处进行完全横断。游泳耐力是功能脊髓修复的一个重要指标。对于游泳耐力,将斑马鱼置于不断增加的水流速度中,直到它们筋疲力尽,并报告它们在耗尽前的时间。为了评估细胞再生,进行组织学检查以分析损伤处胶质和轴突桥的延伸程度。