Ruberg M, Brugg B, Prigent A, Hirsch E, Brice A, Agid Y
INSERM U289, Paris, France.
J Neurochem. 1997 May;68(5):2098-110. doi: 10.1046/j.1471-4159.1997.68052098.x.
A cDNA library of substantia nigra pars compacta from a patient with Parkinson's disease (PD) was differentially screened with probes of normal and parkinsonian substantia nigra enriched in neuronal transcripts. Fifty-eight clones were isolated; 39 were subunits of mitochondrial respiratory complexes I and IV. Parallel screening of a cDNA library derived from normal substantia nigra confirmed differential representation of the transcripts in the substantia nigra pars compacta. In situ hybridization in postmortem brain from parkinsonian and control subjects, with representative complex I and complex IV probes, showed increased labeling, at the cellular level, of the complex I subunit ND1 in neurons of the lateral substantia nigra, where cell death is greatest in PD, but decreased labeling in the medial substantia nigra where fewer cells die. Expression of a complex IV subunit, COXI, increased, however, in both parts of the structure. Increased expression of ND1 and COXI was also observed in nerve growth factor-differentiated PC12 cells undergoing apoptosis induced by tumor necrosis factor-alpha, suggesting that the differential regulation of certain mitochondrial mRNAs may be associated with this form of cell death. This in vitro model of apoptosis is potentially relevant to the death of dopaminergic neurons in PD, because these cells express the tumor necrosis factor-alpha receptor, and neighboring microglial cells in patients synthesize the cytokine.
用富含神经元转录本的正常和帕金森病黑质探针，对一名帕金森病（PD）患者的致密部黑质cDNA文库进行差异筛选。分离出58个克隆；39个是线粒体呼吸复合物I和IV的亚基。对来自正常黑质的cDNA文库进行平行筛选，证实了这些转录本在致密部黑质中的差异表达。用代表性的复合物I和复合物IV探针，对帕金森病患者和对照受试者的死后大脑进行原位杂交，结果显示，在细胞水平上，外侧黑质神经元中复合物I亚基ND1的标记增加，而外侧黑质是PD中细胞死亡最严重的部位；内侧黑质细胞死亡较少，其标记减少。然而，复合物IV亚基COXI在该结构的两个部分中的表达均增加。在经肿瘤坏死因子-α诱导凋亡的神经生长因子分化的PC12细胞中，也观察到ND1和COXI的表达增加，这表明某些线粒体mRNA的差异调节可能与这种细胞死亡形式有关。这种体外凋亡模型可能与PD中多巴胺能神经元的死亡有关，因为这些细胞表达肿瘤坏死因子-α受体，并且患者体内相邻的小胶质细胞会合成细胞因子。
