Elevated alpha-synuclein mRNA levels in individual UV-laser-microdissected dopaminergic substantia nigra neurons in idiopathic Parkinson's disease.
Elevated alpha-synuclein mRNA levels in individual UV-laser-microdissected dopaminergic substantia nigra neurons in idiopathic Parkinson's disease.
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DOI：
10.1093/nar/gkn084
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发表时间：
2008-04
影响因子：
14.9
通讯作者：
Liss B
中科院分区：
文献类型：
--
作者：
Gründemann J;Schlaudraff F;Haeckel O;Liss B
突触前蛋白α-突触核蛋白与包括帕金森病（PD）在内的多种神经退行性疾病有关。在罕见的家族性PD中，已鉴定出α-突触核蛋白基因的因果突变（PARK 1）以及倍增（PARK 4）。在散发性、特发性PD中，α-突触核蛋白的异常蓄积和沉积也可能导致多巴胺能中脑神经元变性，这是PD中临床上最相关的神经元群体。因此，与对照相比，特发性PD患者多巴胺能神经元中α-突触核蛋白表达水平的细胞特异性定量将提供关于α-突触核蛋白对PD病因学贡献的重要信息。然而，许多先前在组织水平上解决这个问题的研究产生了关于α-突触核蛋白表达的不同结果。为了增加特异性，我们开发了一种细胞特异性的mRNA定量方法，该方法还考虑到了个体人类死后脑样本的可变RNA完整性的重要问题。我们证明，PCR扩增子的大小可以混淆定量基因表达分析，特别是部分降解的RNA。通过将优化的紫外激光显微切割和定量RT-PCR技术与合适的PCR检测相结合，我们检测到与对照组相比，特发性PD脑中存活的神经黑色素和酪氨酸羟化酶阳性黑质多巴胺能神经元中α-突触核蛋白mRNA水平显著升高。这些结果加强了α-突触核蛋白基因转录失调在散发性PD中的病理生理作用。
The presynaptic protein α-synuclein is involved in several neurodegenerative diseases, including Parkinson's disease (PD). In rare familial forms of PD, causal mutations (PARK1) as well as multiplications (PARK4) of the α-synuclein gene have been identified. In sporadic, idiopathic PD, abnormal accumulation and deposition of α-synuclein might also cause degeneration of dopaminergic midbrain neurons, the clinically most relevant neuronal population in PD. Thus, cell-specific quantification of α-synuclein expression-levels in dopaminergic neurons from idiopathic PD patients in comparison to controls would provide essential information about contributions of α-synuclein to the etiology of PD. However, a number of previous studies addressing this question at the tissue-level yielded varying results regarding α-synuclein expression. To increase specificity, we developed a cell-specific approach for mRNA quantification that also took into account the important issue of variable RNA integrities of the individual human postmortem brain samples. We demonstrate that PCR –amplicon size can confound quantitative gene-expression analysis, in particular of partly degraded RNA. By combining optimized UV-laser microdissection- and quantitative RT–PCR-techniques with suitable PCR assays, we detected significantly elevated α-synuclein mRNA levels in individual, surviving neuromelanin- and tyrosine hydroxylase-positive substantia nigra dopaminergic neurons from idiopathic PD brains compared to controls. These results strengthen the pathophysiologic role of transcriptional dysregulation of the α-synuclein gene in sporadic PD.
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影响因子：
1.7
作者：
Beyer, Katrin;Humbert, Jordi;Ariza, Aurelio
通讯作者：
Ariza, Aurelio
影响因子：
168.9
作者：
Chartier-Harlin, MC;Kachergus, J;Destée, A
通讯作者：
Destée, A
影响因子：
4.4
作者：
CAMPION, D;MARTIN, C;FREBOURG, T
通讯作者：
FREBOURG, T
影响因子：
9.8
作者：
Chiba-Falek, O;Kowalak, JA;Nussbaum, RL
通讯作者：
Nussbaum, RL
DOI：
10.1073/pnas.0406283101
发表时间：
2004-10-12
影响因子：
11.1
作者：
Chandra, S;Fornai, F;Südhof, TC
通讯作者：
Südhof, TC