Nuclear alpha-synuclein is present in the human brain and is modified in dementia with Lewy bodies.
Nuclear alpha-synuclein is present in the human brain and is modified in dementia with Lewy bodies.
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DOI：
10.1186/s40478-022-01403-x
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发表时间：
2022-07-06
影响因子：
7.1
通讯作者：
中科院分区：
文献类型：
--
作者：
路易体痴呆（DLB）在病理学上定义为大脑神经元内α-突触核蛋白（aSyn）的细胞质积聚。主要是突触前，aSyn已在实验模型中的各种亚细胞区室中报道。事实上，核α-突触核蛋白（aSynNuc）在许多模型中是明显的，其失调与改变的DNA完整性、转录和核稳态有关。然而，aSynNuc在人脑细胞中的存在仍然存在争议，然而人脑aSynNuc及其病理学修饰的测定对于理解突触核蛋白病是必不可少的。在这里，使用多学科的方法，采用免疫组织化学，免疫印迹和质谱（MS），我们确认aSynNuc在死后的脑组织中获得的DLB和对照病例。高度依赖于抗原修复方法，在最佳条件下，在所有检查的病例中，在固定脑切片和分离的核制备物中的皮质神经元和非神经元细胞中观察到核内泛和磷酸-S129阳性aSyn斑点。此外，在神经元和非神经元细胞类型中，与对照相比，DLB病例中核磷酸-S129阳性aSyn免疫反应性的增加是明显的。我们最初的组织学研究确定aSynNuc受表位解蔽方法的影响，但在最佳条件下存在，并且这种存在通过细胞核的分离以及免疫印迹和质谱的组合方法来证实，其中aSynNuc在细胞核中的丰度比细胞质低约10倍。值得注意的是，DLB病例与老年对照的直接比较鉴定了DLB病例的细胞核中增加的pS129和更高分子量物质，表明DLB中aSynNuc的推定致病性修饰。总之，使用多种方法，我们提供了支持aSynNuc在尸检人脑组织中存在的几条证据，值得注意的是，它在DLB中受到可能导致疾病表型的推定致病性修饰。在线版本包含补充材料，可通过10.1186/s40478-022-01403-x获得。
Dementia with Lewy bodies (DLB) is pathologically defined by the cytoplasmic accumulation of alpha-synuclein (aSyn) within neurons in the brain. Predominately pre-synaptic, aSyn has been reported in various subcellular compartments in experimental models. Indeed, nuclear alpha-synuclein (aSynNuc) is evident in many models, the dysregulation of which is associated with altered DNA integrity, transcription and nuclear homeostasis. However, the presence of aSynNuc in human brain cells remains controversial, yet the determination of human brain aSynNuc and its pathological modification is essential for understanding synucleinopathies. Here, using a multi-disciplinary approach employing immunohistochemistry, immunoblot, and mass-spectrometry (MS), we confirm aSynNuc in post-mortem brain tissue obtained from DLB and control cases. Highly dependent on antigen retrieval methods, in optimal conditions, intra-nuclear pan and phospho-S129 positive aSyn puncta were observed in cortical neurons and non-neuronal cells in fixed brain sections and in isolated nuclear preparations in all cases examined. Furthermore, an increase in nuclear phospho-S129 positive aSyn immunoreactivity was apparent in DLB cases compared to controls, in both neuronal and non-neuronal cell types. Our initial histological investigations identified that aSynNuc is affected by epitope unmasking methods but present under optimal conditions, and this presence was confirmed by isolation of nuclei and a combined approach of immunoblotting and mass spectrometry, where aSynNuc was approximately tenfold less abundant in the nucleus than cytoplasm. Notably, direct comparison of DLB cases to aged controls identified increased pS129 and higher molecular weight species in the nuclei of DLB cases, suggesting putative pathogenic modifications to aSynNuc in DLB. In summary, using multiple approaches we provide several lines of evidence supporting the presence of aSynNuc in autoptic human brain tissue and, notably, that it is subject to putative pathogenic modifications in DLB that may contribute to the disease phenotype. The online version contains supplementary material available at 10.1186/s40478-022-01403-x.
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丝氨酸-129上骨料形成蛋白α-核蛋白的磷酸化抑制其DNA弯曲特性。
DOI：
10.1016/j.jbc.2021.101552
发表时间：
2022-03
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Dent SE;King DP;Osterberg VR;Adams EK;Mackiewicz MR;Weissman TA;Unni VK
通讯作者：
Unni VK
DOI：
10.3791/53415
发表时间：
2016-01-05
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
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通讯作者：
Bandopadhyay R
影响因子：
3.3
作者：
Huang, Z.;Xu, Z.;Wu, Y.;Zhou, Y.
通讯作者：
Zhou, Y.
影响因子：
21.3
作者：
Fujiwara, H;Hasegawa, M;Iwatsubo, T
通讯作者：
Iwatsubo, T
影响因子：
12.7
作者：
Koss DJ;Jones G;Cranston A;Gardner H;Kanaan NM;Platt B
通讯作者：
Platt B