Quantification of Endosome and Lysosome Motilities in Cultured Neurons Using Fluorescent Probes

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2017 Jun 2

PMID 28570534

Citations 4

Authors

Fuminori Tsuruta

Tomomi Okajima

Sarasa Yano

Tomoki Chiba

Affiliations

Soon will be listed here.

Abstract

In the brain, membrane trafficking systems play important roles in regulating neuronal functions, such as neuronal morphology, synaptic plasticity, survival, and glial communications. To date, numerous studies have reported that defects in these systems cause various neuronal diseases. Thus, understanding the mechanisms underlying vesicle dynamics may provide influential clues that could aid in the treatment of several neuronal disorders. Here, we describe a method for quantifying vesicle motilities, such as motility distance and rate of movement, using a software plug-in for the ImageJ platform. To obtain images for quantification, we labeled neuronal endosome-lysosome structures with EGFP-tagged vesicle marker proteins and observed the movement of vesicles using a time-lapse microscopy. This method is highly useful and simplify measuring vesicle motility in neurites, such as axons and dendrites, as well as in the soma of both neurons and glial cells. Furthermore, this method can be applied to other cell lines, such as fibroblasts and endothelial cells. This approach could provide a valuable advancement of our understanding of membrane trafficking.

Citing Articles

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Early onset effects of single substrate accumulation recapitulate major features of LSD in patient-derived lysosomes.

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Characterisation of Endocytic Trafficking within Airway Epithelial Cells Using High-Resolution Automated Quantitative Confocal Microscopy.

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Microglial dynamics during brain development.

Okajima T, Tsuruta F Neural Regen Res. 2018; 13(2):222-223.

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