Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, P.O. Box 999, Richland, Washington 99352, USA.
J Proteome Res. 2010 Mar 5;9(3):1496-509. doi: 10.1021/pr901024z.
Parkinson's disease (PD) is characterized by dopaminergic neurodegeneration in the nigrostriatal region of the brain; however, the neurodegeneration extends well beyond dopaminergic neurons. To gain a better understanding of the molecular changes relevant to PD, we applied two-dimensional LC-MS/MS to comparatively analyze the proteome changes in four brain regions (striatum, cerebellum, cortex, and the rest of brain) using a MPTP-induced PD mouse model with the objective to identify potential nigrostriatal-specific and other region-specific protein abundance changes. The combined analyses resulted in the identification of 4,895 nonredundant proteins with at least two unique peptides per protein. The relative abundance changes in each analyzed brain region were estimated based on the spectral count information. A total of 518 proteins were observed with substantial MPTP-induced abundance changes across different brain regions. A total of 270 of these proteins were observed with specific changes occurring either only in the striatum and/or in the rest of the brain region that contains substantia nigra, suggesting that these proteins are associated with the underlying nigrostriatal pathways. Many of the proteins that exhibit changes were associated with dopamine signaling, mitochondrial dysfunction, the ubiquitin system, calcium signaling, the oxidative stress response, and apoptosis. A set of proteins with either consistent change across all brain regions or with changes specific to the cortex and cerebellum regions were also detected. Ubiquitin specific protease (USP9X), a deubiquination enzyme involved in the protection of proteins from degradation and promotion of the TGF-beta pathway, exhibited altered abundance in all brain regions. Western blot validation showed similar spatial changes, suggesting that USP9X is potentially associated with neurodegeneration. Together, this study for the first time presents an overall picture of proteome changes underlying both nigrostriatal pathways and other brain regions potentially involved in MPTP-induced neurodegeneration. The observed molecular changes provide a valuable reference resource for future hypothesis-driven functional studies of PD.
帕金森病（PD）的特征是大脑黑质纹状体区域多巴胺能神经元变性；然而，神经退行性变远远超出了多巴胺能神经元。为了更好地了解与 PD 相关的分子变化，我们应用二维 LC-MS/MS 比较分析了 MPTP 诱导的 PD 小鼠模型中四个脑区（纹状体、小脑、皮质和大脑其余部分）的蛋白质组变化，目的是确定潜在的黑质纹状体特异性和其他区域特异性蛋白质丰度变化。联合分析鉴定了 4895 个非冗余蛋白，每个蛋白至少有 2 个独特肽。根据谱计数信息估计了每个分析脑区的相对丰度变化。总共观察到 518 个蛋白质在不同脑区有明显的 MPTP 诱导的丰度变化。这些蛋白质中有 270 个观察到特定变化，这些变化仅发生在纹状体和/或含有黑质的大脑其余区域，这表明这些蛋白质与潜在的黑质纹状体途径有关。许多表现出变化的蛋白质与多巴胺信号、线粒体功能障碍、泛素系统、钙信号、氧化应激反应和细胞凋亡有关。还检测到一组在所有脑区都有一致变化或仅在皮质和小脑区域有变化的蛋白质。泛素特异性蛋白酶（USP9X）是一种参与保护蛋白质免受降解和促进 TGF-β 途径的去泛素化酶，在所有脑区的丰度都发生了改变。Western blot 验证显示了相似的空间变化，表明 USP9X 可能与神经退行性变有关。总的来说，这项研究首次全面展示了潜在的黑质纹状体途径和其他可能参与 MPTP 诱导神经退行性变的脑区的蛋白质组变化。观察到的分子变化为未来 PD 的假设驱动功能研究提供了有价值的参考资源。
