Imaging fitc-dextran as a reporter for regulated exocytosis

Ofir Klein, Amit Roded, Koret Hirschberg, Mitsunori Fukuda, Stephen J. Galli, Ronit Sagi-Eisenberg

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Regulated exocytosis is a process by which cargo, which is stored in secretory granules (SGs), is released in response to a secretory trigger. Regulated exocytosis is fundamental for intercellular communication and is a key mechanism for the secretion of neurotransmitters, hormones, inflammatory mediators, and other compounds, by a variety of cells. At least three distinct mechanisms are known for regulated exocytosis: full exocytosis, where a single SG fully fuses with the plasma membrane, kiss-and-run exocytosis, where a single SG transiently fuses with the plasma membrane, and compound exocytosis, where several SGs fuse with each other, prior to or after SG fusion with the plasma membrane. The type of regulated exocytosis undertaken by a cell is often dictated by the type of secretory trigger. However, in many cells, a single secretory trigger can activate multiple modes of regulated exocytosis simultaneously. Despite their abundance and importance across cell types and species, the mechanisms that determine the different modes of secretion are largely unresolved. One of the main challenges in investigating the different modes of regulated exocytosis, is the difficulty in distinguishing between them as well as exploring them separately. Here we describe the use of fluorescein isothiocyanate (FITC)-dextran as an exocytosis reporter, and live cell imaging, to differentiate between the different pathways of regulated exocytosis, focusing on compound exocytosis, based on the robustness and duration of the exocytic events.

Original languageEnglish
Article numbere57936
JournalJournal of Visualized Experiments
Volume2018
Issue number136
DOIs
Publication statusPublished - 2018 Jun 20

Keywords

  • Biology
  • Compound exocytosis
  • FITC-dextran
  • Issue 136
  • Live cell imaging
  • Lysosome-related organelles
  • Pinocytosis
  • Regulated exocytosis

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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  • Cite this

    Klein, O., Roded, A., Hirschberg, K., Fukuda, M., Galli, S. J., & Sagi-Eisenberg, R. (2018). Imaging fitc-dextran as a reporter for regulated exocytosis. Journal of Visualized Experiments, 2018(136), [e57936]. https://doi.org/10.3791/57936