Neurotrauma is among the main causes of human disability and mortality. Nerve injury impairs not only neurons but also causes death of satellite glial cells remote from the injury site. We studied the dynamics of expression of different proapoptotic proteins (E2F1, p53, caspase 3) in the dorsal root ganglia (DRG) of a rat after sciatic nerve transection. TUNEL staining and immunoblotting were used for analysis of cell apoptosis and axotomy-induced biochemical changes. Apoptosis of glial cells was observed at 24 h after sciatic nerve transection and increased on day 7, when apoptosis of some neurons only started. The earliest proapoptotic event in the injured DRG was overexpression of transcription factor E2F1 at 4 h after sciatic nerve transection. This preceded the induction of p53 and cleavage of caspase 3 at 24-h post-axotomy. The nerve injury marker amyloid precursor protein and the nerve regeneration marker GAP-43 were overexpressed in DRG on day 7 after sciatic nerve transection. We also developed a novel fluorescence method for differential visualization of the rat DRG and nerves by means of double staining with propidium iodide and Hoechst 33342 that impart red and blue-green fluorescence, respectively. The present experiments showed that glial cells remote from the nerve transection site were more vulnerable to axotomy than DRG neurons. E2F1 and p53 may be considered promising molecular targets for development of potential neuroprotective agents.

神经创伤是人类残疾和死亡的主要原因之一。神经损伤不仅会损害神经元，还会导致远离损伤部位的卫星神经胶质细胞死亡。我们研究了坐骨神经横断后大鼠背根神经节 (DRG) 中不同促凋亡蛋白 (E2F1、p53、半胱天冬酶 3) 的表达动态。TUNEL 染色和免疫印迹用于分析细胞凋亡和轴突切开诱导的生化变化。在坐骨神经横断后 24 小时观察到神经胶质细胞的凋亡，并在第 7 天增加，此时一些神经元的凋亡才开始。受损 DRG 中最早的促凋亡事件是坐骨神经横断后 4 小时转录因子 E2F1 的过度表达。这在轴突切开后 24 小时诱导 p53 和裂解 caspase 3 之前。神经损伤标志物淀粉样前体蛋白和神经再生标志物 GAP-43 在坐骨神经横断后第 7 天在 DRG 中过表达。我们还开发了一种新的荧光方法，通过使用碘化丙啶和 Hoechst 33342 分别赋予红色和蓝绿色荧光的双重染色，对大鼠 DRG 和神经进行差异可视化。目前的实验表明，远离神经横断部位的神经胶质细胞比 DRG 神经元更容易受到轴突切断术的影响。E2F1 和 p53 可能被认为是开发潜在神经保护剂的有希望的分子靶点。我们还开发了一种新的荧光方法，通过使用碘化丙啶和 Hoechst 33342 分别赋予红色和蓝绿色荧光的双重染色，对大鼠 DRG 和神经进行差异可视化。目前的实验表明，远离神经横断部位的神经胶质细胞比 DRG 神经元更容易受到轴突切断术的影响。E2F1 和 p53 可能被认为是开发潜在神经保护剂的有希望的分子靶点。我们还开发了一种新的荧光方法，通过使用碘化丙啶和 Hoechst 33342 分别赋予红色和蓝绿色荧光的双重染色，对大鼠 DRG 和神经进行差异可视化。目前的实验表明，远离神经横断部位的神经胶质细胞比 DRG 神经元更容易受到轴突切断术的影响。E2F1 和 p53 可能被认为是开发潜在神经保护剂的有希望的分子靶点。