Department of Neurology, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Department of Neurology, Osaka University Graduate School of Medicine, Suita, Japan.
J Neuropathol Exp Neurol. 2020 Apr 1;79(4):370-377. doi: 10.1093/jnen/nlaa003.
Alterations of RNA metabolism caused by mutations in RNA-binding protein genes, such as transactivating DNA-binding protein-43 (TDP-43) and fused in sarcoma (FUS), have been implicated in the pathogenesis of amyotrophic lateral sclerosis (ALS). Unlike the accumulation of TDP43, which is accepted as a pathological hall mark of sporadic ALS (sALS), FUS pathology in sALS is still under debate. Although immunoreactive inclusions of FUS have been detected in sALS patients previously, the technical limitation of signal detection, including the necessity of specific antigen retrieval, restricts our understanding of FUS-associated ALS pathology. In this study, we applied a novel detection method using a conventional antigen retrieval technique with Sudan Black B treatment to identify FUS-positive inclusions in sALS patients. We classified pathological motor neurons into 5 different categories according to the different aggregation characteristics of FUS and TDP-43. Although the granular type was more dominant for inclusions with TDP-43, the skein-like type was more often observed in FUS-positive inclusions, suggesting that these 2 proteins undergo independent aggregation processes. Moreover, neurons harboring FUS-positive inclusions demonstrated substantially reduced expression levels of dynactin-1, a retrograde motor protein, indicating that perturbation of nucleocytoplasmic transport is associated with the formation of cytoplasmic inclusions of FUS in sALS.
RNA 结合蛋白基因突变导致的 RNA 代谢改变,如反式激活 DNA 结合蛋白 43(TDP-43)和肉瘤融合(FUS),与肌萎缩侧索硬化症(ALS)的发病机制有关。与 TDP43 的积累不同,TDP43 的积累被认为是散发性 ALS(sALS)的病理标志,而 FUS 病理学在 sALS 中仍存在争议。尽管先前在 sALS 患者中检测到 FUS 的免疫反应性包含物,但信号检测的技术限制,包括对特定抗原检索的必要性,限制了我们对 FUS 相关 ALS 病理学的理解。在这项研究中,我们应用了一种新的检测方法,使用常规抗原检索技术结合苏丹黑 B 处理来鉴定 sALS 患者中的 FUS 阳性包含物。我们根据 FUS 和 TDP-43 的不同聚集特征将病理运动神经元分为 5 种不同类别。尽管 TDP-43 包含物的颗粒型更为优势,但 FUS 阳性包含物中更常观察到线丝型,表明这两种蛋白质经历独立的聚集过程。此外,含有 FUS 阳性包含物的神经元表现出动力蛋白-1(一种逆行运动蛋白)的表达水平显著降低,表明核质转运的扰动与 sALS 中 FUS 细胞质包含物的形成有关。