小鼠巨噬细胞内游离钙离子变化介导的吞噬作用

为评估小鼠巨噬细胞吞噬死亡细胞时胞质内游离钙离子的变化。实验使用F luo-3标记巨噬细胞内钙离子和碘化丙碇对死亡细胞核染色,观察吞噬过程中细胞内钙离子的变化和显示巨噬细胞的吞噬功能,检测含死亡细胞的巨噬细胞内荧光密度图像。利用激光扫描共聚焦显微镜检测钙离子的释放。在缺钙的溶液中,可见巨噬细胞接触死细胞时细胞内钙离子快速地聚集和增高。在吞噬体形成时,巨噬细胞内钙离子上升到较高的水平。快速上升后,当吞噬小泡消化时,细胞内游离钙下降,随后钙离子恢复到低水平。研究显示伴随着吞噬小泡中红色荧光的死细胞的出现和消失,巨噬细胞内出现一系列钙离子变化的图像。提示巨噬细胞内钙离子改变在细胞吞噬作用中具有一定的作用。

Cytosolic Free Clcium Fluctuation Mediates Phagocytosis in Mouse Macrophages

Cytosolic free calciu.m(Ca~(2 )]i) change was assessed in single adherent mouse macrophages during phagocytosis for dead cells.To determine visualizing Ca~(2 ) change during phagocytosis and indirectly show phagocytic function of macrophages to dead cells,intracellular Ca~(2 )]i of macrophages were loaded with the fluorescent dye Fluo-3/AM and dead cells were labeled with Propidium Iodide(PI),we examined images pattern of the fluorescence intensity in macrophages of containing dead cells.A discharge of calcium was verified by confocal scanning laser microscopy(CSLM).In lacking Ca~(2 ) medium,upon contact with dead cells,rapid assemble and increase ofCa~(2 )]i was observed in macrophage.Intracellular Ca~(2 )]i rose to a higher level during initial phagosome formation.After a rapid rise,free Ca~(2 )was decreased in digestion of phagocytotic vesicle;there-after the Ca~(2 )]i returned to the baseline.Our study directly indicates that accompany presenting and disappearing of dead cell of red fluorescence in the phagosomal vesicle,a series of images pattern of Ca~(2 )]i change can be really exhibited in macrophages.It is suggested that Ca~(2 )]i fluctuation has a certainty in macrophages during the phagocytosis