Boston Children's Hospital and Harvard Medical School, Boston, MA 02115, USA.
Cell Rep. 2013 Jul 25;4(2):385-401. doi: 10.1016/j.celrep.2013.06.018. Epub 2013 Jul 11.
Microglia are resident immune cells of the CNS that are activated by infection, neuronal injury, and inflammation. Here, we utilize flow cytometry and deep RNA sequencing of acutely isolated spinal cord microglia to define their activation in vivo. Analysis of resting microglia identified 29 genes that distinguish microglia from other CNS cells and peripheral macrophages/monocytes. We then analyzed molecular changes in microglia during neurodegenerative disease activation using the SOD1(G93A) mouse model of amyotrophic lateral sclerosis (ALS). We found that SOD1(G93A) microglia are not derived from infiltrating monocytes, and that both potentially neuroprotective and toxic factors, including Alzheimer's disease genes, are concurrently upregulated. Mutant microglia differed from SOD1(WT), lipopolysaccharide-activated microglia, and M1/M2 macrophages, defining an ALS-specific phenotype. Concurrent messenger RNA/fluorescence-activated cell sorting analysis revealed posttranscriptional regulation of microglia surface receptors and T cell-associated changes in the transcriptome. These results provide insights into microglia biology and establish a resource for future studies of neuroinflammation.
小胶质细胞是中枢神经系统中的固有免疫细胞，可被感染、神经元损伤和炎症激活。在这里，我们利用流式细胞术和急性分离的脊髓小胶质细胞的深度 RNA 测序来定义它们在体内的激活。对静止小胶质细胞的分析确定了 29 个基因，这些基因将小胶质细胞与其他中枢神经系统细胞和外周巨噬细胞/单核细胞区分开来。然后，我们使用 SOD1(G93A)肌萎缩侧索硬化 (ALS)小鼠模型分析了小胶质细胞在神经退行性疾病激活过程中的分子变化。我们发现，SOD1(G93A)小胶质细胞不是由浸润的单核细胞衍生而来的，包括阿尔茨海默病基因在内的潜在神经保护和毒性因子同时上调。突变型小胶质细胞与 SOD1(WT)、脂多糖激活的小胶质细胞和 M1/M2 巨噬细胞不同，定义了一种 ALS 特异性表型。同时进行的信使 RNA/荧光激活细胞分选分析揭示了小胶质细胞表面受体的转录后调控和转录组中 T 细胞相关变化。这些结果提供了对小胶质细胞生物学的深入了解，并为神经炎症的未来研究建立了资源。
