Optical analysis of cell membrane dynamics and recycling processes in cultured human ethmoidal cells using a fluorescent dye FM4-64.

概要

論文の詳細を見る
The styrylpyridinium dyes, which are nontoxic and water-soluble membrane probes, have been used both for investigating the mechanisms of activity-dependent membrane recycling and for visualizing the endocytosis phase in a wide range of species. In this study, we visualized cell membrane-internalization and transport in cultured human ethmoidal cells using a fluorescent probe FM4-64 and laser scanning confocal microscopy. After treatment for 5min at 30°C, FM4-64 labeling became apparent in the plasma membrane of ciliated cells. The labeling was pronounced on the apical side corresponding to the area where numerous cilia assembled. A different pattern was observed in non-ciliated cells where the plasma membrane was weakly fluorescent on their basolateral side. Using time-lapse conf ocal microscopy, we were able to follow vesicle internalization and augmented membrane turnover in some of the ciliated cells triggered by various concentrations of histamine (10-3M to 10-5M). The fluorescent labeling of these cells began to rise until 10 minutes concomitant with distinctly visualized beating cilia. We have shown that this dye serves as a sensitive vital reporter for membrane dynamics, detecting such events as endosome to membrane transport during receptor-mediated endocytosis in vitro.
日本鼻科学会の論文