Department of Neurology, Philipps-University Marburg, Germany.
Neurosci Lett. 2012 Jan 17;507(1):10-5. doi: 10.1016/j.neulet.2011.11.035. Epub 2011 Dec 1.
Parkinson's disease (PD) is a chronic neurodegenerative disorder characterized by progressive loss of dopaminergic (DA) neurons of the substantia nigra pars compacta with unknown aetiology. 6-Hydroxydopamine (6-OHDA) treatment of neuronal cells is an established in vivo model for mimicking the effect of oxidative stress found in PD brains. We examined the effects of 6-OHDA treatment on human neuroblastoma cells (SH-SY5Y) and primary mesencephalic cultures. Using a reverse arbitrarily primed polymerase chain reaction (RAP-PCR) approach we generated reproducible genetic fingerprints of differential expression levels in cell cultures treated with 6-OHDA. Of the resulting sequences, 23 showed considerable homology to known human coding sequences. The results of the RAP-PCR were validated by reverse transcription PCR, real-time PCR and, for selected genes, by Western blot analysis and immunofluorescence. In four cases, [tomoregulin-1 (TMEFF-1), collapsin response mediator protein 1 (CRMP-1), neurexin-1, and phosphoribosylaminoimidazole synthetase (GART)], a down-regulation of mRNA and protein levels was detected. Further studies will be necessary on the physiological role of the identified proteins and their impact on pathways leading to neurodegeneration in PD.
帕金森病（PD）是一种慢性神经退行性疾病，其特征是黑质致密部多巴胺能（DA）神经元进行性丧失，病因不明。6-羟多巴胺（6-OHDA）处理神经元细胞是一种公认的体内模型，可模拟 PD 大脑中发现的氧化应激效应。我们研究了 6-OHDA 处理对人神经母细胞瘤细胞（SH-SY5Y）和原代中脑神经培养物的影响。使用反向任意引物聚合酶链反应（RAP-PCR）方法，我们生成了 6-OHDA 处理细胞培养物中差异表达水平的可重复遗传指纹图谱。在产生的序列中，有 23 个与已知的人类编码序列具有相当大的同源性。RAP-PCR 的结果通过逆转录 PCR、实时 PCR 以及对选定基因的 Western blot 分析和免疫荧光进行了验证。在四种情况下，[TMEFF-1（tomoregulin-1）、CRMP-1（collapsin response mediator protein 1）、neurexin-1 和 GART（phosphoribosylaminoimidazole synthetase）]，检测到 mRNA 和蛋白水平下调。还需要进一步研究鉴定出的蛋白质的生理作用及其对 PD 中导致神经退行性变途径的影响。
