摘要
目的观察CD2AP在小鼠大脑中的表达情况,探究其是否会发生相分离,以及对小胶质细胞吞噬和迁移功能的影响。方法使用免疫荧光染色法研究CD2AP蛋白在WT小鼠和AD模型小鼠大脑中的表达和定位情况;在转染CD2AP的293T细胞中检测液滴的液态动力学;使用CD2AP重组蛋白观察液滴的形成;体外培养HMC3细胞并转染,通过划痕实验和细胞吞噬实验,检测对照组和CD2AP组细胞迁移率和吞噬的差异。结果CD2AP在小胶质细胞中呈高表达,呈液滴状分布。胞内过表达CD2AP后出现了具有流动性的液滴。CD2AP体外重组蛋白在不同拥挤剂的作用下不同时间点均能形成液滴。CD2AP过表达组细胞迁移率明显增高(P<0.05),但吞噬能力下降(P<0.05)。结论CD2AP具有液-液相分离的能力,并且影响小胶质细胞的吞噬和迁移能力。
Objective To observe the expression of CD2AP in the mouse brain,investigate whether it undergoes phase separation,and assess its effect on the phagocytic and migratory functions of microglia.Methods Immunofluorescence staining was used to study the expression and localization of CD2AP protein in the brains of wild-type(WT)mice and Alzheimer's disease(AD)model mice.The liquid dynamics of droplets was detected in 293T cells transfected with CD2AP.The formation of droplets was observed using CD2AP recombinant protein.HMC3 cells were cultured and transfected in vitro,and the differences in cell mobility and phagocytosis between control and CD2AP groups were assessed using scratch assay and cell phagocytosis assay.Results CD2AP was highly expressed in microglia,exhibiting a droplet-like distribution.Intracellular overexpression of CD2AP resulted in droplets exhibiting fluidity.In vitro,recombinant protein CD2AP formed droplets at various time points in the presence of different crowding agents.Cell scratch assay and phagocytosis assays showed that the CD2AP overexpression group had significantly higher cell migration(P<0.05)but decreased phagocytosis(P<0.05).Conclusion CD2AP has the ability to undergo liquid-liquid phase separation and affects the phagocytic and migratory abilities of microglia.
作者
陆炯彤
陆星宇
李胜男
曾凯星
张佳昊
崔理立
LU Jiongtong;LU Xingyu;LI Shengnan;ZENG Kaixing;ZHANG Jiahao;CUI Lili(Institute of Neurology,Guangdong Key Laboratory of Age-Related Cardiac and Cerebral Diseases,Affiliated Hospital of Guangdong Medical University,Zhanjiang 524002,China;The Marine Biomedical Research Institute of Guangdong,School of Ocean and Tropical Medicine,Guangdong Medical University,Zhanjiang 524002,China)
出处
《广东医科大学学报》
2025年第4期379-388,共10页
Journal of Guangdong Medical University
基金
国家自然科学基金(8237052644)。
