体外评估小鼠小胶质细胞对突触体的吞噬和降解的实验方案。

Matera Alessandro, Compagnion Anne-Claire, Paolicelli Rosa Chiara

University of Lausanne, Department of Biomedical Sciences, 1005 Lausanne, Switzerland.

STAR Protoc. 2025 Jul 10;6(3):103936. doi: 10.1016/j.xpro.2025.103936.

Microglia, the innate immune cells of the central nervous system, refine neuronal circuitries both during brain development and in neurodegenerative diseases. Here, we present a protocol to independently assess the engulfment and degradation of synaptosomes by murine microglia, both in fixed and live samples, using confocal imaging. We describe steps for isolating primary microglia, preparing and conjugating synaptosomes with pHrodo, and performing an uptake and degradation assay. We then detail procedures for analyzing synaptosome uptake and degradation using ImageJ software. For complete details on the use and execution of this protocol, please refer to Matera, Compagnion, et al..

小胶质细胞是中枢神经系统的固有免疫细胞，在大脑发育和神经退行性疾病过程中都会优化神经回路。在此，我们提供了一种方案，可通过共聚焦成像，在固定样本和活样本中独立评估小鼠小胶质细胞对突触小体的吞噬和降解。我们描述了分离原代小胶质细胞、用pHrodo标记突触小体并进行摄取和降解分析的步骤。然后详细介绍了使用ImageJ软件分析突触小体摄取和降解的程序。有关本方案使用和执行的完整详细信息，请参阅马特拉、康帕尼翁等人的相关内容。