Ibarburu Sofía, Trias Emiliano, Lago Natalia, Peluffo Hugo, Barreto-Núñez Romina, Varela Valentina, Beckman Joseph S, Barbeito Luis
Laboratorio de Neurodegeneración, Institut Pasteur de Montevideo, Montevideo, Uruguay.
Neuroimmunomodulation. 2017;24(3):143-153. doi: 10.1159/000480639. Epub 2017 Nov 1.
We aimed to determine the potential of aberrant glial cells (AbAs) isolated from the spinal cord of adult SOD1G93A symptomatic rats to induce gliosis and neuronal damage following focal transplantation into the lumbar spinal cord of wild-type rats.
AbAs were obtained from the spinal cords of SOD1G93A symptomatic rats. One hundred thousand cells were injected using a glass micropipette into the lumbar spinal cords (L3-L5) of syngeneic wild-type adult rats. Equal volumes of culture medium or wild-type neonatal microglia were used as controls. Seven days after transplantation, immunohistochemistry analysis was carried out using astrocytic and microglia cell markers. Transplanted SOD1G93A AbAs were recognized by specific antibodies to human SOD1 (hSOD1) or misfolded human SOD1.
Seven days after transplantation, AbAs were mainly detected in the medial region of the lumbar ventral horn as a well-limited cell cluster formed at the site of injection by their immunoreactivity to either misfolded SOD1 or normally folded hSOD1. Compared with controls, transplanted AbAs were surrounded by marked microgliosis and reactive astrocytes. Marked microgliosis was observed to extend bilaterally up to the cervical cord. Motor neurons close to AbA transplants were surrounded by activated glial cells and displayed ubiquitin aggregation.
AbAs bearing mutant SOD1G93A have the potential to induce neuroinflammation along the spinal cord and incipient damage to the motor neurons. The emergence of AbAs during amyotrophic lateral sclerosis pathogenesis may therefore be a mechanism to boost neuroinflammation and spread motor neuron damage along the neuroaxis.
我们旨在确定从成年SOD1G93A症状性大鼠脊髓中分离出的异常胶质细胞（AbAs）在局灶性移植到野生型大鼠腰脊髓后诱导胶质增生和神经元损伤的可能性。
从SOD1G93A症状性大鼠的脊髓中获取AbAs。使用玻璃微量移液器将10万个细胞注射到同基因野生型成年大鼠的腰脊髓（L3-L5）中。等量的培养基或野生型新生小胶质细胞用作对照。移植后7天，使用星形胶质细胞和小胶质细胞标志物进行免疫组织化学分析。移植的SOD1G93A AbAs通过针对人SOD1（hSOD1）或错误折叠的人SOD1的特异性抗体进行识别。
移植后7天，AbAs主要在腰腹角内侧区域被检测到，通过其对错误折叠的SOD1或正常折叠的hSOD1的免疫反应性，在注射部位形成一个界限清楚的细胞簇。与对照组相比，移植的AbAs被明显的小胶质细胞增生和反应性星形胶质细胞包围。观察到明显的小胶质细胞增生双侧延伸至颈髓。靠近AbA移植部位的运动神经元被活化的胶质细胞包围，并显示泛素聚集。
携带突变型SOD1G93A的AbAs有潜力诱导脊髓神经炎症和运动神经元的早期损伤。因此，在肌萎缩侧索硬化症发病过程中AbAs的出现可能是促进神经炎症和沿神经轴传播运动神经元损伤的一种机制。
