Investigating mast cell secretory granules; from biosynthesis to exocytosis

Nurit P. Azouz, Mitsunori Fukuda, Marc E. Rothenberg, Ronit Sagi-Eisenberg

    Research output: Contribution to journalArticlepeer-review

    9 Citations (Scopus)

    Abstract

    Mast Cells (MC) are secretory cells of the immune system that accomplish their physiological and pathological functions by releasing pre-formed and newly synthesized allergic, inflammatory and immunoregulatory mediators. MCs’ mediators affect multiple tissues and organs culminating in allergic and immune responses. The synthesis, storage and release of the MC mediators are highly regulated. The pre-formed mediators are packed in cytoplasmic secretory granules (SG) that fuse with the plasma membrane and release their content by regulated exocytosis. We present a protocol, based on the co-expression of a gene of interest with a reporter gene that is targeted to the SGs and is released in a regulated fashion alongside the endogenous SG mediators. The protocol enables high resolution four dimensional confocal analyses of the MC SGs and monitoring their timeline from biogenesis to triggered exocytosis. Thus, using this protocol for screening genes of interest for their phenotypic and functional impact allows deciphering the molecular mechanisms that govern the biogenesis and exocytosis of the MC SGs and identifying the regulators involved. Thereby, further insights into the cellular mechanisms that account for MCs function in health and disease should be provided.

    Original languageEnglish
    Article numbere52505
    JournalJournal of Visualized Experiments
    Issue number95
    DOIs
    Publication statusPublished - 2015 Jan 26

    Keywords

    • Confocal imaging
    • Exocytosis
    • Immunology
    • Issue 95
    • Live cell imaging
    • Mast cells
    • Rab5
    • Reporter gene
    • Secretory granules
    • Transfection

    ASJC Scopus subject areas

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

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