Department of Developmental Biology, Washington University School of Medicine.
Department of Developmental Biology, Washington University School of Medicine; Center of Regenerative Medicine, Washington University School of Medicine;
J Vis Exp. 2021 Nov 12(177). doi: 10.3791/63240.
Due to their renowned regenerative capacity, adult zebrafish are a premier vertebrate model to interrogate mechanisms of innate spinal cord regeneration. Following complete transection of their spinal cord, zebrafish extend glial and axonal bridges across severed tissue, regenerate neurons proximal to the lesion, and regain their swim capacities within 8 weeks of injury. Recovery of swim function is thus a central readout for functional spinal cord repair. Here, we describe a set of behavioral assays to quantify zebrafish motor capacity inside an enclosed swim tunnel. The goal of these methods is to provide quantifiable measurements of swim endurance and swim behavior in adult zebrafish. For swim endurance, zebrafish are subjected to a constantly increasing water current velocity until exhaustion, and time at exhaustion is reported. For swim behavior assessment, zebrafish are subjected to low current velocities and swim videos are captured with a dorsal view of the fish. Percent activity, burst frequency, and time spent against the water current provide quantifiable readouts of swim behavior. We quantified swim endurance and swim behavior in wild-type zebrafish before injury and after spinal cord transection. We found that zebrafish lose swim function after spinal cord transection and gradually regain that capacity between 2 and 6 weeks post-injury. The methods described in this study could be applied to neurobehavioral, musculoskeletal, skeletal muscle regeneration, and neural regeneration studies in adult zebrafish.
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。在其脊髓完全横断后,斑马鱼会在切断的组织上延伸胶质和轴突桥,在损伤部位近端再生神经元,并在损伤后 8 周内恢复游泳能力。因此,游泳功能的恢复是衡量脊髓功能修复的一个重要指标。在这里,我们描述了一套在封闭的游泳隧道中量化斑马鱼运动能力的行为检测方法。这些方法的目的是为成年斑马鱼的游泳耐力和游泳行为提供可量化的测量。对于游泳耐力,斑马鱼会在不断增加的水流速度下直到力竭,记录力竭时间。对于游泳行为评估,斑马鱼会在低水流速度下进行,并用鱼的背视图拍摄游泳视频。活动百分比、爆发频率和对抗水流的时间提供了游泳行为的可量化指标。我们在损伤前和脊髓横断后对野生型斑马鱼进行了游泳耐力和游泳行为的量化。我们发现,斑马鱼在脊髓横断后失去了游泳功能,并在损伤后 2 至 6 周逐渐恢复该功能。本研究中描述的方法可应用于成年斑马鱼的神经行为、肌肉骨骼、骨骼肌再生和神经再生研究。