Center for Neuroscience, Shantou University Medical College, Shantou, People's Republic of China.
PLoS One. 2012;7(12):e52376. doi: 10.1371/journal.pone.0052376. Epub 2012 Dec 21.
The cell neural adhesion molecule contactin-2 plays a key role in axon extension and guidance, fasciculation, and myelination during development. We thus asked, whether contactin-2 is also important in nervous system regeneration after trauma. In this study, we used an adult zebrafish spinal cord transection model to test the functions of contactin-2 in spinal cord regeneration. The expression patterns of contactin-2 at different time points after spinal cord injury were studied at the mRNA level by qPCR and in situ hybridization, and contactin-2 protein levels and immunohistological localization were detected by Western blot and immunofluorescence analyses, respectively. Contactin-2 mRNA and protein levels were increased along the central canal at 6 days and 11 days after spinal cord injury, suggesting a requirement for contactin-2 in spinal cord regeneration. Co-localization of contactin-2 and islet-1 (a motoneuron marker) was observed in spinal cords before and after injury. To further explore the functions of contactin-2 in regeneration, an anti-sense morpholino was used to knock down the expression of contactin-2 protein by application at the time of injury. Motion analysis showed that inhibition of contactin-2 retarded the recovery of swimming functions when compared to standard control morpholino. Anterograde and retrograde tracing at 6 weeks after injury showed that knock down of contactin-2 inhibited axonal regrowth from NMLF neurons beyond lesion site. The combined observations indicate that contactin-2 contributes to locomotor recovery and successful regrowth of axons after spinal cord injury in adult zebrafish.
细胞黏附分子接触蛋白-2 在轴突延伸和导向、纤维束形成以及发育过程中的髓鞘形成中发挥着关键作用。因此,我们提出这样一个问题:接触蛋白-2 在创伤后神经系统再生中是否也很重要。在本研究中,我们使用成年斑马鱼脊髓横断模型来测试接触蛋白-2 在脊髓再生中的功能。通过 qPCR 和原位杂交技术在 mRNA 水平研究了脊髓损伤后不同时间点接触蛋白-2 的表达模式,通过 Western blot 和免疫荧光分析分别检测了接触蛋白-2 蛋白水平和免疫组织化学定位。在脊髓损伤后 6 天和 11 天,接触蛋白-2 mRNA 和蛋白水平沿中央管增加,表明接触蛋白-2 在脊髓再生中是必需的。在损伤前后的脊髓中观察到接触蛋白-2 和胰岛 1(运动神经元标志物)的共定位。为了进一步探索接触蛋白-2 在再生中的功能,在损伤时应用反义 morpholino 敲低接触蛋白-2 蛋白的表达。运动分析表明,与标准对照 morpholino 相比,抑制接触蛋白-2 会延迟游泳功能的恢复。损伤后 6 周的顺行和逆行示踪显示,接触蛋白-2 的敲低抑制了 NMLF 神经元内从损伤部位以外的轴突再生。综合观察表明,接触蛋白-2 有助于成年斑马鱼脊髓损伤后的运动功能恢复和轴突的成功再生。