Dissecting SNARE-Mediated Exocytosis in RBL-2H3 Mast Cells

Overview

Journal Methods Mol Biol

Date 2025 Jan 13

PMID 39806152

Authors

Pratikshya Adhikari

Hao Xu

Affiliations

Soon will be listed here.

Abstract

SNARE-dependent mast cell (MC) exocytosis causes the release of a wide variety of mediators with important physiological/pathological consequences. Unlike synaptic transmission in the brain, which relies primarily on one set of exocytic SNAREs (i.e., Syntaxin1, SNAP-25, and VAMP2), MCs produce a multitude of exocytic SNAREs that can form a minimum of 8 distinct sets of fusogenic trans-SNARE complexes. Here we describe the genetic approaches we have developed to dissect the specific roles of these SNAREs in RBL-2H3 cells, a widely utilized model for studying MC signaling and exocytosis.

References

Rao K, Brown M . Mast cells: multifaceted immune cells with diverse roles in health and disease. Ann N Y Acad Sci. 2008; 1143:83-104. DOI: 10.1196/annals.1443.023. View

Wezel A, Quax P, Kuiper J, Bot I . The role of mast cells in atherosclerosis. Hamostaseologie. 2014; 35(2):113-20. DOI: 10.5482/HAMO-14-08-0034. View

Manikandan J, Kothandaraman N, Hande M, Pushparaj P . Deciphering the structure and function of FcεRI/mast cell axis in the regulation of allergy and anaphylaxis: a functional genomics paradigm. Cell Mol Life Sci. 2011; 69(12):1917-29. PMC: 11114762. DOI: 10.1007/s00018-011-0886-0. View

Chapman D, Irvin C . Mechanisms of airway hyper-responsiveness in asthma: the past, present and yet to come. Clin Exp Allergy. 2015; 45(4):706-19. PMC: 4386586. DOI: 10.1111/cea.12506. View

Netzer N, Kupper T, Voss H, Eliasson A . The actual role of sodium cromoglycate in the treatment of asthma--a critical review. Sleep Breath. 2012; 16(4):1027-32. DOI: 10.1007/s11325-011-0639-1. View

Platt M . Pharmacotherapy for allergic rhinitis. Int Forum Allergy Rhinol. 2014; 4 Suppl 2:S35-40. DOI: 10.1002/alr.21381. View

Krop M, Ozunal Z, Chai W, de Vries R, Fekkes D, Bouhuizen A . Mast cell degranulation mediates bronchoconstriction via serotonin and not via renin release. Eur J Pharmacol. 2010; 640(1-3):185-9. DOI: 10.1016/j.ejphar.2010.04.058. View

Kalesnikoff J, Galli S . New developments in mast cell biology. Nat Immunol. 2008; 9(11):1215-23. PMC: 2856637. DOI: 10.1038/ni.f.216. View

Theoharides T, Bondy P, Tsakalos N, Askenase P . Differential release of serotonin and histamine from mast cells. Nature. 1982; 297(5863):229-31. DOI: 10.1038/297229a0. View

10.

Puri N, Roche P . Mast cells possess distinct secretory granule subsets whose exocytosis is regulated by different SNARE isoforms. Proc Natl Acad Sci U S A. 2008; 105(7):2580-5. PMC: 2268179. DOI: 10.1073/pnas.0707854105. View

11.

Tiwari N, Wang C, Brochetta C, Ke G, Vita F, Qi Z . VAMP-8 segregates mast cell-preformed mediator exocytosis from cytokine trafficking pathways. Blood. 2008; 111(7):3665-74. DOI: 10.1182/blood-2007-07-103309. View

12.

Frank S, Thon K, Bischoff S, Lorentz A . SNAP-23 and syntaxin-3 are required for chemokine release by mature human mast cells. Mol Immunol. 2011; 49(1-2):353-8. DOI: 10.1016/j.molimm.2011.09.011. View

13.

Baram D, Adachi R, Medalia O, Tuvim M, Dickey B, Mekori Y . Synaptotagmin II negatively regulates Ca2+-triggered exocytosis of lysosomes in mast cells. J Exp Med. 1999; 189(10):1649-58. PMC: 2193646. DOI: 10.1084/jem.189.10.1649. View

14.

Moon T, Befus A, Kulka M . Mast cell mediators: their differential release and the secretory pathways involved. Front Immunol. 2014; 5:569. PMC: 4231949. DOI: 10.3389/fimmu.2014.00569. View

15.

Fasshauer D, Sutton R, Brunger A, Jahn R . Conserved structural features of the synaptic fusion complex: SNARE proteins reclassified as Q- and R-SNAREs. Proc Natl Acad Sci U S A. 1998; 95(26):15781-6. PMC: 28121. DOI: 10.1073/pnas.95.26.15781. View

16.

Sudhof T, Rothman J . Membrane fusion: grappling with SNARE and SM proteins. Science. 2009; 323(5913):474-7. PMC: 3736821. DOI: 10.1126/science.1161748. View

17.

Woska Jr J, Gillespie M . Small-interfering RNA-mediated identification and regulation of the ternary SNARE complex mediating RBL-2H3 mast cell degranulation. Scand J Immunol. 2010; 73(1):8-17. DOI: 10.1111/j.1365-3083.2010.02471.x. View

18.

Paumet F, Le Mao J, Martin S, GALLI T, David B, Blank U . Soluble NSF attachment protein receptors (SNAREs) in RBL-2H3 mast cells: functional role of syntaxin 4 in exocytosis and identification of a vesicle-associated membrane protein 8-containing secretory compartment. J Immunol. 2000; 164(11):5850-7. DOI: 10.4049/jimmunol.164.11.5850. View

19.

Tiwari N, Wang C, Brochetta C, Scandiuzzi L, Hong W, Blank U . Increased formation of VAMP-3-containing SNARE complexes in mast cells from VAMP-8 deficient cells. appetite. Inflamm Res. 2009; 58 Suppl 1:13-4. DOI: 10.1007/s00011-009-0645-y. View

20.

Brochetta C, Suzuki R, Vita F, Soranzo M, Claver J, Madjene L . Munc18-2 and syntaxin 3 control distinct essential steps in mast cell degranulation. J Immunol. 2013; 192(1):41-51. PMC: 3905451. DOI: 10.4049/jimmunol.1301277. View