Characterization of Mast Cell Secretory Granules and Their Cell Biology

Azouz, Nurit P.; Hammel, Ilan; Sagi‐Eisenberg, Ronit

doi:10.1089/dna.2014.2543

Cited by 34 publications

(29 citation statements)

References 49 publications

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“…injection of fluorochrome-labeled avidin (sulforhodamine 101-coupled avidin [Av.SRho]) into the ear pinna dermis 10 to 30 minutes before performing 2-photon microscopy (15). During MC degranulation, which results in fusion of secretory granule membranes with the plasma membrane (16,17), externalized granule matrices are rapidly bound by soluble fluorescent Av.SRho probe present in the microenvironment, allowing imaging by high-resolution confocal and in vivo 2-photon microscopy of such exteriorized granule structures (15,18). While this method is useful for identifying exteriorized MC granule structures, it requires (a) that MCs are activated to degranulate, and (b) using specific MC reporter mice (e.g., Mcpt5-EYFP mice; see refs.…”

Section: Resultsmentioning

confidence: 99%

Imaging protective mast cells in living mice during severe contact hypersensitivity

Reber¹,

Sibilano²,

Starkl³

et al. 2017

JCI Insight

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Section: Resultsmentioning

confidence: 99%

Imaging protective mast cells in living mice during severe contact hypersensitivity

Reber¹,

Sibilano²,

Starkl³

et al. 2017

JCI Insight

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“…Specifically, we showed that NPY-mRFP is targeted to the SGs in BMMCs and RBL cultured mast cells, and is released in a regulated fashion alongside the endogenous mediators, thus allowing us to monitor and quantify exocytosis selectively in the genetically manipulated cells (15)(16)(17). Furthermore, the pH insensitivity of mRFP fluorescence allows visualization of the acidic SGs, permitting four-dimensional analyses of the SG dynamics in correlation with the exocytic process (5). Using this method to confirm FIGURE 1.…”

Section: Rab12 Knockdown Potentiates Whereas Ca Rab12 Inhibits Regumentioning

confidence: 92%

“…A major fraction of these mediators, including vasoactive amines such as histamine or serotonin, multiple proteases, and lysosomal hydrolases, is prestored in lysosome-related secretory granules (SGs) and released by exocytosis in response to cell trigger by a variety of external cues (reviewed in Refs. 4,5). The latter include the immunological trigger that is mediated by binding and subsequent cross-linking of allergen-directed IgE Abs to FcεRI, the high-affinity receptor for IgE (reviewed in Refs.…”

mentioning

confidence: 99%

Rab12 Regulates Retrograde Transport of Mast Cell Secretory Granules by Interacting with the RILP–Dynein Complex

Efergan

Azouz

Klein

et al. 2016

The Journal of Immunology

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Secretory granule (SG) transport is a critical step in regulated exocytosis including degranulation of activated mast cells. The latter process results in the release of multiple inflammatory mediators that play key roles in innate immunity, as well as in allergic responses. In this study, we identified the small GTPase Rab12 as a novel regulator of mast cell SG transport, and we provide mechanistic insights into its mode of action. We show that Rab12 is activated in a stimulus-dependent fashion and promotes microtubule-dependent retrograde transport of the SGs in the activated cells. We also show that this minus end transport of the SGs is mediated by the RILP–dynein complex and identify RILP as a novel effector of Rab12. Finally, we show that Rab12 negatively regulates mast cell degranulation. Taken together, our results identify Rab12 as a novel regulator of mast cell responses and disclose for the first time, to our knowledge, the mechanism of retrograde transport of the mast cell SGs.

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“…In line with this premise, mast cell (MC) models have been developed for studying the molecular basis of allergy, a family of diseases, which share their onset upon the release of inflammatory mediators from activated MCs. The latter, including vasoactive amines such as histamine, proteases such as tryptase, proteoglycans such as heparin, lysosomal enzymes such as β‐hexosaminidase and some cytokines, are prestored in cytoplasmic secretory granules (SGs) and released by exocytosis following MC activation by external triggers . This initial event of exocytosis is followed by the de novo synthesis and release of a large array of biologically potent substances, including metabolites of arachidonic acid, multiple cytokines, and chemokines .…”

Section: Introductionmentioning

confidence: 99%

RBL cells as models for in vitro studies of mast cells and basophils

Falcone

Wan

Barwary

et al. 2018

Immunological Reviews

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Since their establishment in 1981, RBL-2H3 cells have been widely used as a mast cell (MC) model. Their ability to be easily grown in culture in large amounts, their responsiveness to FcεRI-mediated triggers and the fact that they can be genetically manipulated, have provided advantages over primary MCs, in particular for molecular studies relying on genetic screening. Furthermore, the ability to generate clones that stably express proteins of interest, for example, a human receptor, have marked the RBL cells as an attractive MC model for drug screening. Indeed, 3 RBL reporter cell lines (RS-ATL8, NFAT-DsRed, and NPY-mRFP) have been generated providing useful models for drug and allergen screening. Similarly, RBL cells stably expressing the human MrgprX2 receptor provide a unique paradigm for analyzing ligand interactions and signaling pathways of the unique human receptor. Finally, transient co-transfections of RBL cells allow functional genomic analyses of MC secretion by combining library screening with simultaneous expression of a reporter for exocytosis. RBL cells thus comprise powerful tools for the study of intracellular membrane trafficking and exocytosis and the detection of allergens, vaccine safety studies and diagnosis of allergic sensitization. Their recent uses as an investigative tool are reviewed here.

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Characterization of Mast Cell Secretory Granules and Their Cell Biology

Cited by 34 publications

References 49 publications

Imaging protective mast cells in living mice during severe contact hypersensitivity

Imaging protective mast cells in living mice during severe contact hypersensitivity

Rab12 Regulates Retrograde Transport of Mast Cell Secretory Granules by Interacting with the RILP–Dynein Complex

RBL cells as models for in vitro studies of mast cells and basophils

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