Evaluation of phagocytic function and analysis of antigen-processing mechanism of microglia in ALS

首先，我们用免疫组织化学方法评估ALS小鼠模型的小胶质行为和脊髓损伤。我们发现SOD1G85R转基因小鼠在症状期脊髓中小胶质细胞摄取或包围了SOD1聚集体。因此，为什么我们认为这些小胶质细胞不能完全消化SOD1聚集体时反映了“吞噬抑制”的状态？接下来，为了量化聚集体被小胶质细胞吸收的情况，我们建立了测量吞噬细胞吸收荧光珠的系统。利用该系统，我们将表达SOD1G85R的小胶质细胞与野生型小胶质细胞进行了比较，但没有发现差异。我们证实SOD1WTor g85rfibrils的聚集体在添加到培养基中时被纳入小胶质细胞。然后我们将sod1g85r突变体通过吸附sod1g85r制成SOD1聚集模拟物，并检测野生型或表达sod1g85r的小胶质细胞对SOD1聚集模拟物的吸收能力，但没有发现差异。

First, we evaluatebyimmunohistochemistry the microglialbehaviorinspinalcord lesion of ALS mouse model. We found microglia uptaked, or surrounded the SOD1 aggregates in the spinal cord of the SOD1G85R transgenic mouse at symptomatic stage. Therefore wehy pothes ized that these microglia reflected the state of"frustrated phagocytosis"when microglia could not digest SOD1 aggregates fully. Next, in order to quantify th uptake of aggregates into microglia, we establishedthe system tomeasure the fluorescent beads incorporated into the phagocytes Using the system, we compared the SOD1G85R expressing-microglia with wild-type ones, but no difference was found. We confirmedtheaggregates ofthe SOD1WTor G85Rfibrilswere incorpotrated into microglia when they were added into medium. Then we made the mutant SOD1 aggregate-mimics by adsorb SOD1G85Rinto beads andexamined the ability ofwild-type orSOD1G85R-expressing microglia t incorporate them, butno difference was found.