EAAT1 and EAAT2 immunoreactivity in ALS and pathology of SOD1 (G93A) transgenic mice

我们使用EAAT1和EAAT2抗体对肌萎缩性侧索硬化症（ALS）患者的脊髓进行免疫组织化学研究，以检测EAAT1和EAAT2免疫反应性与前角神经元变性之间的关系。在对照组中，脊髓灰质被抗体密集免疫染色，而白质通常没有免疫染色。在ALS患者中，EAAT1的免疫反应性在灰质中保存得相对较好。相比之下，前角EAAT2的免疫反应性与前角细胞神经元丢失程度相关：在轻度神经元缺失患者中，前角被抗体密集免疫染色，而在严重神经元丢失患者中，EAAT2的表达明显降低。因此，我们证明了EAAT1和EAAT2在ALS不同进展阶段的免疫反应性存在差异，这是该疾病的病理机制的一个特征。我们还对G93A SOD1基因突变的转基因小鼠脊髓进行了免疫组织化学研究，使用诱导型一氧化氮合酶（iNOS）和硝基酪氨酸（NT）抗体来表征免疫反应性的时间和地形分布，从而阐明了运动系统氧化损伤增加在神经退行性过程中的可能作用。对照组小鼠在任何年龄均未出现iNOS或NT阳性免疫反应。相比之下，在转基因小鼠中，在24周（早期症状前阶段），前角神经元偶有iNOS和NT免疫染色；在28周时（症状前晚期），它们的免疫染色并不罕见；在32周（症状早期）和35周（终末期）时，在增生的反应性星形胶质细胞和前角细胞体细胞中经常观察到iNOS和NT阳性免疫反应。前角神经元中iNOS和NT阳性免疫反应性的选择性定位提示氧化应激可能参与了该转基因小鼠运动神经元变性的病理机制。少

We immunohistochemically investigated the spinal cord of patients with amyotrophic lateral sclerosis (ALS), using EAAT1 and EAAT2 antibodies to examine relationships between EAAT1 and EAAT2 immunoreactivity and degeneration of anterior horn neurons. In controls, spinal cord gray matter was densely immunostained by antibodies, whereas the white matter was generally not immunostained. In ALS patients, EAAT1 immunoreactivity was relatively well-preserved in the gray mater. In contrast, EAAT2 immunoreactivity in anterior horns correlated with the degree of neuronal loss of anterior horn cells : in the patients with mild neuronal depletion, anterior horns were densely immunostained by the antibody, whereas in the patients with severe neuronal loss, EAAT2 expression was markedly reduced. Thus, we demonstrate a difference in EAAT1 and EAAT2 immunoreactivity in different stages of progression in ALS, as a feature of the pathomechanism of this disease. We also performed an immunohistochemical s … More tudy of the spinal cords of transgenic mice with a G93A mutant SOD1 gene, using antibodies to inducible nitric oxide synthase (iNOS) and nitrotyrosine (NT) to characterize the temporal and topographic distribution of the immunoreactivity, thus illuminating the possible role of increased oxidative damage to the motor system in the neurodegenerative process. The control mice showed no positive iNOS or NT immunoreactivity at any age. In contrast, in the transgenic mice, at 24 weeks (early presymptomatic stage), the anterior horn neurons were occasionally immunostained for iNOS and NT ; at 28 weeks (late presymptomatic stage), they were not uncommonly immunostained ; at 32 weeks (early symptomatic stage) and 35 weeks (end-stage), positive iNOS and NT immunoreactivity was frequently observed in proliferated reactive astrocytes as well as in the somata of the anterior horn cells. The selective localization of positive iNOS and NT immunoreactivity in the anterior horn neurons suggests that oxidative stress may be involved in the pathomechanism of degeneration of motor neurons in this transgenic mice. Less