Elucidation of the formation mechanism of neuronal cytoplasmic inclusions in motor neuron diseases

阐明运动神经元疾病中神经元细胞质包涵体的形成机制

• 批准号: 06454277
• 负责人: MIZUSAWA Hidehiro
• 金额: $ 4.42万
• 依托单位: Tokyo Medical and Dental University (1996)University of Tsukuba (1994-1995)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-06454277/
• 关键词: ubiquitin; skein-like inclusion; Lewy body-like hyaline inclusion; Bunina body; round inclusion; neurofilament; lysosome; motor neuron disease; Zn superoxide dismutase; ubiguitin; round inclusion; Lewy小体様; Zn SOD; NOS; cathepsin D

Motor neuron diseases (MNDs) including amyotrophic lateral sclerosis (ALS) have characteristic inclusions such as Lewy body-like hyaline inclusion (LBH), skein-like inclusion (SLI) and Bunina body (BB), although the pathogeneses of MNDs remain unknown. The investigation of the formation pathomechanism of these specific inclusions would provide important clues to elucidate the pathogenesis of MNDs.SLI was essentially a bundle of filaments slightly thicker than neurofilament. SLIs existed singularly or aggregates variously forming networks of bundles, typical skeins and larger inclusions. Single of a few SLIs often appeared similar to BBs particularly when SLIs are cut crossly or obliquely. However, they were distinct because SLIs were ubiquitin-positive and cystatin C-negative while BBs were ubiquitin-negative and cystatin C-positive. Large "SLIs" composed of many bundles usually contained many free filaments associated with granular material which were reminiscent of granule-associated … More thick filaments of round inclusions in sporadic ALS.SLI were negative for neurofilament and Cu/Zu SOD.By ubiquitin immunocytochemistry, SLIs were found in most cases on spradic ALS while similar inclusions were rarely reported in other diseases. Therefore, SLIs have diagnontic value for sporadic ALS.Ultrastructurally some SLIs were observed within double membrane structure probably lysosomes although cathepsin D immunoreactivity of SLIs were not outstanding from cytoplasmic immunoreaction. SLIs were almost negative for ubiquitin C terminal hydrolase or proteasome indicating that ubiquitin system involved in SLI metabolism might be defective and lysosomal system might be also involved.LBHI which was observed in most cases of familial ALS with posterior column degeneration contains neurofilament particularly in the halo while there were also thicker filaments with granular material. They were often round and occasionally cord-like in shape with quite homogeneous content and smooth outline. LBHI was immunocytochemically positive for Cu/Zn SOD and mutations of Cu/Zn SOD gene were found in this type of familial ALS.LBHI may be related to oxydant stress which is possibly a pathomechanism of MND.In rare occasions, round inclusion (RI) similar to LBHI was found in sporadic ALS,particularly in cases with rapid clinical course. RIs were composed mainly of thick filaments with granular material. Hematoxylin and eosin staining as well as ubiqitin immunohistochemistry showed RIs were irregular in shape and inhomogeneous or filamentous in content even if they appeared to have distinct core and halo. RIs were usually not immunoreactive for neurofilament of Cu/Zn SOD.RI often contained bundles of thick filaments which were indistinguishable from SLI.These findings indicate that RI is distinct from LBHI but closely related to so-called large aggregated form of SLI. Less

包括肌萎缩侧索硬化症（ALS）在内的运动神经元病（MND）具有特征性包涵体，如Lewy小体样透明包涵体（LBH）、绞样包涵体（SLI）和Bunina小体（BB），但MND的发病机制尚不清楚。SLI实质上是一束比神经丝稍粗的纤维束，其结构与神经纤维束的结构相似。SLIs以单一或聚集的形式存在，形成束、典型的束和较大的包裹体的网络。少数SLI中的单个SLI通常与BB相似，特别是当SLI交叉或倾斜切割时。然而，它们是不同的，因为SLI是泛素阳性和半胱氨酸蛋白酶抑制剂C阴性，而BB是泛素阴性和半胱氨酸蛋白酶抑制剂C阳性。由许多纤维束组成的大的“SLI”通常含有许多与颗粒物质有关的游离丝，这使人联想到与颗粒物质有关的纤维束。 ...更多信息 SLI对神经丝和Cu/Zu SOD均呈阴性反应，泛素免疫组化结果显示，SLI多见于散发性ALS，而在其他疾病中未见类似的包涵体。超微结构上，SLI的组织蛋白酶D免疫反应在胞浆中不明显，但在双膜结构中可能存在溶酶体。SLI几乎不表达泛素C末端水解酶或蛋白酶体，说明参与SLI代谢的泛素系统可能存在缺陷，溶酶体系统也可能参与SLI的代谢。LBHI见于大多数伴有后柱变性的家族性ALS患者，其内含有神经丝，尤其是在晕圈内，但也有较粗的纤维和颗粒状物质。它们通常是圆形的，偶尔也是索状的，内容非常均匀，轮廓光滑。LBHI为Cu/Zn SOD免疫细胞化学阳性，在该型家族性ALS中发现Cu/Zn SOD基因突变，LBHI可能与氧化应激有关，氧化应激可能是MND的病理机制之一，在极少数情况下，在散发性ALS中发现类似LBHI的圆形包涵体（RI），尤其是在病程较短的病例中。RI主要由粗丝和颗粒状物质组成。苏木精-伊红染色和泛素免疫组化显示RI形状不规则，内容物不均匀或丝状，即使它们看起来有明显的核心和晕。RI通常不表达Cu/Zn SOD的神经丝，RI常含有与SLI难以区分的粗纤维束，这些结果表明RI不同于LBHI，但与所谓的大聚集型SLI密切相关。少

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