Sackler Faculty of Medicine, Tel Aviv University

3 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.

Developmental Biology

Isolating and Analyzing Cells of the Pancreas Mesenchyme by Flow Cytometry

Alona Epshtein¹, Lina Sakhneny¹, Limor Landsman¹

¹Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University

Here, we describe a method for the isolation of cells in the pancreas microenvironment from embryonic, neonatal and adult mouse tissue, focusing on the isolation of mesenchymal cells. This method allows profiling of cell gene expression and protein secretion in order to elucidate the extrinsic signals that regulate pancreas development, function, and tumorigenesis.

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.