Martin L J
Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2196, USA.
J Neuropathol Exp Neurol. 1999 May;58(5):459-71. doi: 10.1097/00005072-199905000-00005.
The mechanisms for neurodegeneration in amyotrophic lateral sclerosis (ALS) are not understood. We found that motor neuron degeneration in ALS structurally resembles apoptosis. The progression of neuronal death is divisible into 3 sequential stages: chromatolysis, somatodendritic attrition, and apoptosis. In ALS spinal cord anterior horn and motor cortex, DNA fragmentation is detectable in situ and in gels and is internucleosomal, occurring in the presence of DNA fragmentation factor-45/40 activation and increased caspase-3 activity. By immunoblotting, changes occur in the subcellular distribution of cell death proteins that would promote apoptosis. In selectively vulnerable CNS regions in ALS compared with controls, the proapoptotic proteins Bax and Bak are elevated in the mitochondrial-enriched membrane compartment, but are reduced or unchanged in the cytosol. In contrast, the antiapoptotic protein Bcl-2 is decreased in the mitochondrial-enriched membrane compartment of vulnerable regions in ALS, but is increased in the cytosol, whereas Bcl-xL levels are unchanged in both subcellular compartments. Coimmunoprecipitation experiments showed that Bax-Bax interactions are greater in the mitochondrial-enriched membrane compartment of ALS motor cortex compared with controls, whereas Bax-Bcl-2 interactions are lower in the membrane compartment of ALS motor cortex compared with controls. We conclude that a PCD mechanism, involving cytosol-to-membrane and membrane-to-cytosol redistribution of cell death proteins and caspase-3 activation, participates in the pathogenesis of ALS.
肌萎缩侧索硬化症(ALS)中神经退行性变的机制尚不清楚。我们发现,ALS中的运动神经元变性在结构上类似于细胞凋亡。神经元死亡的进程可分为3个连续阶段:染色质溶解、体树突萎缩和细胞凋亡。在ALS脊髓前角和运动皮层中,可原位及凝胶中检测到DNA片段化,且为核小体间的片段化,其发生伴随着DNA片段化因子-45/40的激活及caspase-3活性增加。通过免疫印迹法发现,促进细胞凋亡的细胞死亡蛋白的亚细胞分布发生了变化。与对照组相比,在ALS中选择性易损的中枢神经系统区域,促凋亡蛋白Bax和Bak在线粒体丰富的膜区室中升高,但在细胞质中减少或不变。相反,抗凋亡蛋白Bcl-2在ALS易损区域线粒体丰富的膜区室中减少,但在细胞质中增加,而Bcl-xL在两个亚细胞区室中的水平均未改变。免疫共沉淀实验表明,与对照组相比,ALS运动皮层线粒体丰富的膜区室中Bax-Bax相互作用更强,而与对照组相比,ALS运动皮层膜区室中Bax-Bcl-2相互作用更低。我们得出结论,一种程序性细胞死亡(PCD)机制,涉及细胞死亡蛋白在细胞质与膜之间以及膜与细胞质之间的重新分布和caspase-3激活,参与了ALS的发病机制。