Murtinheira Fernanda, Belo João, Boasinha Ana Sofia, Robalo Tiago T, Torres Vukosava M, Pinto Francisco R, Pimenta Constança, Nascimento Patricia, Rodrigues Mario S, Herrera Federico
BioISI - Instituto de Biosistemas e Ciências Integrativas, Faculdade de Ciências da Universidade de Lisboa, 1749-016, Lisbon, Portugal; Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016, Lisboa, Portugal.
BioISI - Instituto de Biosistemas e Ciências Integrativas, Faculdade de Ciências da Universidade de Lisboa, 1749-016, Lisbon, Portugal; Departamento de Física, Faculdade de Ciências, Universidade de Lisboa, 1749-016, Lisboa, Portugal.
Arch Biochem Biophys. 2025 Oct;772:110569. doi: 10.1016/j.abb.2025.110569. Epub 2025 Jul 25.
Autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSACS) is a movement disorder caused by loss-of-function mutations in the sacsin gene. The most common hallmark of this disease is the disruption of intermediate filament networks in cells as diverse as neurons, kidney cells, fibroblasts, astroglia and microglia. Intermediate filaments are the main filaments responsible for the mechanical and viscoelastic properties of cells and tissues, but these have never been investigated in the context of ARSACS. Here, we analyzed the consequences of sacsin loss on the mechanical functions of astroglial-like C6 cells. The phenotype of C6 cells was analyzed by immunocytochemistry, electron microscopy, mass spectrometry, atomic force microscopy and motility/proliferation assays. C6 cells presented an abnormal cytoskeletal and organelle distribution, global proteome alterations linked to cell motility and mechanics, a significant decrease in cell elasticity in the cytoplasm, and a striking reduction in cell motility. These mechanical alterations in glial-like cells could be especially relevant for neuroinflammation and glial scar formation upon CNS injury. Our results support a possible role for alterations in glial functions in ARSACS and provide new tools for understanding the glial-specific mechanisms involved in this movement disorder.
夏尔沃-萨格奈常染色体隐性痉挛性共济失调（ARSACS）是一种由Sacsin基因功能丧失突变引起的运动障碍。这种疾病最常见的特征是在神经元、肾细胞、成纤维细胞、星形胶质细胞和小胶质细胞等多种细胞中，中间丝网络遭到破坏。中间丝是负责细胞和组织机械及粘弹性特性的主要细丝，但从未在ARSACS的背景下对其进行过研究。在此，我们分析了Sacsin缺失对星形胶质细胞样C6细胞机械功能的影响。通过免疫细胞化学、电子显微镜、质谱分析、原子力显微镜以及运动性/增殖分析对C6细胞的表型进行了分析。C6细胞呈现出异常的细胞骨架和细胞器分布、与细胞运动性和力学相关的整体蛋白质组改变、细胞质中细胞弹性显著降低以及细胞运动性显著减少。胶质细胞样细胞的这些机械改变可能与中枢神经系统损伤后的神经炎症和胶质瘢痕形成特别相关。我们的结果支持胶质细胞功能改变在ARSACS中可能发挥的作用，并为理解这种运动障碍所涉及的胶质细胞特异性机制提供了新工具。
