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Graduate School of Life Sciences, Tohoku University

2 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Investigating Mast Cell Secretory Granules; from Biosynthesis to Exocytosis

Nurit P. Azouz^1,2, Mitsunori Fukuda³, Marc E. Rothenberg², Ronit Sagi-Eisenberg¹

¹Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University, ²Division of Allergy and Immunology, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, ³Laboratory of Membrane Trafficking Mechanisms, Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University

The goal of the present protocol was to develop a method that will allow functional genomic analyses of mast cell secretion. The protocol is based on quantitative assessment of the release of a fluorescent reporter gene cotrasfected with the gene of interest and real time analyses of the secretory granule's morphology.

JoVE Journal

Imaging FITC-dextran as a Reporter for Regulated Exocytosis

Ofir Klein¹, Amit Roded¹, Koret Hirschberg², Mitsunori Fukuda³, Stephen J. Galli⁴, Ronit Sagi-Eisenberg¹

¹Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University, ²Department of Pathology, Sackler Faculty of Medicine, Tel Aviv University, ³Laboratory of Membrane Trafficking Mechanisms, Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University, ⁴Departments of Pathology and of Microbiology and Immunology and Sean N. Parker Center for Allergy and Asthma Research, School of Medicine, Stanford University

Here we detail a method for live cell imaging of regulated exocytosis. This method utilizes FITC-dextran, which accumulates in lysosome-related organelles, as a reporter. This simple method also allows distinguishing between different modes of regulated exocytosis in cells that are difficult to manipulate genetically.