Analyze Diet
The EMBO journal1993; 12(5); 1811-1817; doi: 10.1002/j.1460-2075.1993.tb05829.x

Focal exocytosis by eosinophils–compound exocytosis and cumulative fusion.

Abstract: We have investigated the granule fusion events during exocytosis in horse eosinophils by time-resolved patch-clamp capacitance measurements. Stimulation with intracellular GTP gamma S leads to a stepwise capacitance increase by 4.0 +/- 0.9 pF. At GTP gamma S concentrations < 20 microM the step size distribution is in agreement with the granule size distribution in resting cells. Above 80 microM the number of steps is reduced and very large steps occur. The total capacitance increase, however, is unaffected. These results show that at high GTP gamma S concentrations granule--granule fusion occurs inside the cell forming large compound granules, which then fuse with the plasma membrane (compound exocytosis). The electrical equivalent circuit of the cell during degranulation indicates the formation of a degranulation sac by cumulative fusion events. Fusion of the first granule with the plasma membrane induces fusion of further granules with this granule directing the release of all the granular material to the first fusion pore. The physiological function of eosinophils is the killing of parasites. Compound exocytosis and cumulative fusion enable the cells to focus the release of cytotoxic proteins to well defined target regions and prevent uncontrolled diffusion of this material, which would damage intact host cells.
Publication Date: 1993-05-01 PubMed ID: 8491174PubMed Central: PMC413400DOI: 10.1002/j.1460-2075.1993.tb05829.xGoogle Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article focuses on the process and role of exocytosis in horse eosinophils, a type of white blood cell. The find suggests that high concentrations of GTP gamma S invoke a form of granule fusion within the cell, leading to formation of large compound granules that merge with the plasma membrane in a process known as ‘compound exocytosis’. This focused release of cytotoxic proteins helps in combating parasites.

Investigation of Granule Fusion Events

  • The research was aimed to understand granule fusion events during exocytosis in horse eosinophils. The process was observed using time-resolved patch-clamp capacitance measurements.
  • Stimulation of eosinophils with intracellular GTP gamma S caused a stepwise increase in capacitance by approximately 4.0 ± 0.9 pF.
  • When GTP gamma S concentrations were less than 20 microM, the step size distribution corresponded with the granule size distribution in resting cells.

Granule Fusion in High GTP gamma S Concentrations

  • In situations where GTP gamma S concentrations exceeded 80 microM, the number of steps were reduced, while very large steps occurred.
  • Interestingly, the total capacitance increase remained unaffected in such conditions.
  • This observation indicated that higher GTP gamma S concentrations resulted in granule-granule fusion occurring within the cell, leading to the formation of large compound granules.
  • These large compound granules were then observed to fuse with the plasma membrane, a process termed ‘compound exocytosis’.

Formation of Degranulation Sac & Role of Eosinophils

  • The experiment further revealed that the cell’s electrical equivalent circuit during degranulation indicated the formation of a degranulation sac through cumulative fusion events.
  • This sequence suggests that the initial fusion of the first granule with the plasma membrane triggers the fusion of additional granules with the initial granule.
  • This mechanism leads to the release of all granular material to the first fusion pore.
  • Functionally, eosinophils are implicated in the elimination of parasites. Through compound exocytosis and cumulative fusion processes, these cells can focus the release of cytotoxic proteins in specific target areas.
  • This targeted release prevents unintentional diffusion of cytotoxic materials that could potentially harm intact host cells.

Cite This Article

APA
Scepek S, Lindau M. (1993). Focal exocytosis by eosinophils–compound exocytosis and cumulative fusion. EMBO J, 12(5), 1811-1817. https://doi.org/10.1002/j.1460-2075.1993.tb05829.x

Publication

ISSN: 0261-4189
NlmUniqueID: 8208664
Country: England
Language: English
Volume: 12
Issue: 5
Pages: 1811-1817

Researcher Affiliations

Scepek, S
  • Abteilung Molekulare Zellforschung, Max-Planck-Institut für medzinische Forschung, Heidelberg, Germany.
Lindau, M

    MeSH Terms

    • Animals
    • Cell Membrane / metabolism
    • Cytoplasmic Granules / metabolism
    • Eosinophils / cytology
    • Eosinophils / metabolism
    • Eosinophils / physiology
    • Exocytosis
    • Guanosine 5'-O-(3-Thiotriphosphate) / metabolism
    • Horses
    • In Vitro Techniques
    • Membrane Fusion
    • Membrane Potentials

    References

    This article includes 21 references
    1. Röhlich P, Anderson P, Uvnäs B. Electron microscope observations on compounds 48-80-induced degranulation in rat mast cells. Evidence for sequential exocytosis of storage granules.. J Cell Biol 1971 Nov;51(21):465-83.
      pubmed: 4107023doi: 10.1083/jcb.51.2.465google scholar: lookup
    2. Lindau M, Nüsse O, Bennett J, Cromwell O. The membrane fusion events in degranulating guinea pig eosinophils.. J Cell Sci 1993 Jan;104 ( Pt 1):203-10.
      pubmed: 8449998doi: 10.1242/jcs.104.1.203google scholar: lookup
    3. Lawson D, Fewtrell C, Raff MC. Localized mast cell degranulation induced by concanavalin A-sepharose beads. Implications for the Ca2+ hypothesis of stimulus-secretion coupling.. J Cell Biol 1978 Nov;79(2 Pt 1):394-400.
      pubmed: 569156doi: 10.1083/jcb.79.2.394google scholar: lookup
    4. Glauert AM, Butterworth AE, Sturrock RF, Houba V. The mechansim of antibody-dependent, eosinophil-mediated damage to schistosomula of Schistosoma mansoni in vitro: a study by phase-contrast and electron microscopy.. J Cell Sci 1978 Dec;34:173-92.
      pubmed: 748337doi: 10.1242/jcs.34.1.173google scholar: lookup
    5. Dvorak AM, Galli SJ, Morgan E, Galli AS, Hammond ME, Dvorak HF. Anaphylactic degranulation of guinea pig basophilic leukocytes. I. Fusion of granule membranes and cytoplasmic vesicles formation and resolution of degranulation sacs.. Lab Invest 1981 Feb;44(2):174-91.
      pubmed: 6162057
    6. Tai PC, Spry CJ. The mechanisms which produce vacuolated and degranulated eosinophils.. Br J Haematol 1981 Oct;49(2):219-26.
    7. Chandler DE, Bennett JP, Gomperts B. Freeze-fracture studies of chemotactic peptide-induced exocytosis in neutrophils: evidence for two patterns of secretory granule fusion.. J Ultrastruct Res 1983 Feb;82(2):221-32.
      pubmed: 6827649doi: 10.1016/s0022-5320(83)90055-2google scholar: lookup
    8. Henderson WR, Chi EY, Jörg A, Klebanoff SJ. Horse eosinophil degranulation induced by the ionophore A23187. Ultrastructure and role of phospholipase A2.. Am J Pathol 1983 Jun;111(3):341-9.
      pubmed: 6407328
    9. Breckenridge LJ, Almers W. Currents through the fusion pore that forms during exocytosis of a secretory vesicle.. Nature 1987 Aug 27-Sep 2;328(6133):814-7.
      pubmed: 2442614doi: 10.1038/328814a0google scholar: lookup
    10. Drust DS, Creutz CE. Aggregation of chromaffin granules by calpactin at micromolar levels of calcium.. Nature 1988 Jan 7;331(6151):88-91.
      pubmed: 2963226doi: 10.1038/331088a0google scholar: lookup
    11. Lindau M, Neher E. Patch-clamp techniques for time-resolved capacitance measurements in single cells.. Pflugers Arch 1988 Feb;411(2):137-46.
      pubmed: 3357753doi: 10.1007/BF00582306google scholar: lookup
    12. Fidler N, Fernandez JM. Phase tracking: an improved phase detection technique for cell membrane capacitance measurements.. Biophys J 1989 Dec;56(6):1153-62.
      pubmed: 2611329doi: 10.1016/S0006-3495(89)82762-6google scholar: lookup
    13. Nüsse O, Lindau M, Cromwell O, Kay AB, Gomperts BD. Intracellular application of guanosine-5'-O-(3-thiotriphosphate) induces exocytotic granule fusion in guinea pig eosinophils.. J Exp Med 1990 Mar 1;171(3):775-86.
      pubmed: 2137856doi: 10.1084/jem.171.3.775google scholar: lookup
    14. Alvarez de Toledo G, Fernandez JM. Compound versus multigranular exocytosis in peritoneal mast cells.. J Gen Physiol 1990 Mar;95(3):397-409.
      pubmed: 2324701doi: 10.1085/jgp.95.3.397google scholar: lookup
    15. Gomperts BD. GE: a GTP-binding protein mediating exocytosis.. Annu Rev Physiol 1990;52:591-606.
    16. Spruce AE, Breckenridge LJ, Lee AK, Almers W. Properties of the fusion pore that forms during exocytosis of a mast cell secretory vesicle.. Neuron 1990 May;4(5):643-54.
      pubmed: 2344404doi: 10.1016/0896-6273(90)90192-igoogle scholar: lookup
    17. Gorvel JP, Chavrier P, Zerial M, Gruenberg J. rab5 controls early endosome fusion in vitro.. Cell 1991 Mar 8;64(5):915-25.
      pubmed: 1900457doi: 10.1016/0092-8674(91)90316-qgoogle scholar: lookup
    18. Balch WE. Small GTP-binding proteins in vesicular transport.. Trends Biochem Sci 1990 Dec;15(12):473-7.
      pubmed: 2077687doi: 10.1016/0968-0004(90)90301-qgoogle scholar: lookup
    19. Lindau M. Time-resolved capacitance measurements: monitoring exocytosis in single cells.. Q Rev Biophys 1991 Feb;24(1):75-101.
      pubmed: 2047522doi: 10.1017/s0033583500003279google scholar: lookup
    20. Vogel SS, Zimmerberg J. Proteins on exocytic vesicles mediate calcium-triggered fusion.. Proc Natl Acad Sci U S A 1992 May 15;89(10):4749-53.
      pubmed: 1584814doi: 10.1073/pnas.89.10.4749google scholar: lookup
    21. McLaren DJ, Mackenzie CD, Ramalho-Pinto FJ. Ultrastructural observations on the in vitro interaction between rat eosinophils and some parasitic helminths (Schistosoma mansoni, Trichinella spiralis and Nippostrongylus brasiliensis).. Clin Exp Immunol 1977 Oct;30(1):105-18.
      pubmed: 564249

    Citations

    This article has been cited 35 times.
    1. Chan CY, Faragalla Y, Wu LG. Illuminating membrane structural dynamics of fusion and endocytosis with advanced light imaging techniques.. Biochem Soc Trans 2022 Aug 31;50(4):1157-1167.
      doi: 10.1042/BST20210263pubmed: 35960003google scholar: lookup
    2. Ge L, Shin W, Arpino G, Wei L, Chan CY, Bleck CKE, Zhao W, Wu LG. Sequential compound fusion and kiss-and-run mediate exo- and endocytosis in excitable cells.. Sci Adv 2022 Jun 17;8(24):eabm6049.
      doi: 10.1126/sciadv.abm6049pubmed: 35714180google scholar: lookup
    3. Germic N, Hosseini A, Yousefi S, Karaulov A, Simon HU. Regulation of eosinophil functions by autophagy.. Semin Immunopathol 2021 Jun;43(3):347-362.
      doi: 10.1007/s00281-021-00860-1pubmed: 34019141google scholar: lookup
    4. Klein O, Sagi-Eisenberg R. Anaphylactic Degranulation of Mast Cells: Focus on Compound Exocytosis.. J Immunol Res 2019;2019:9542656.
      doi: 10.1155/2019/9542656pubmed: 31011586google scholar: lookup
    5. Huang CH, Moser T. Ca(2+) Regulates the Kinetics of Synaptic Vesicle Fusion at the Afferent Inner Hair Cell Synapse.. Front Cell Neurosci 2018;12:364.
      doi: 10.3389/fncel.2018.00364pubmed: 30386210google scholar: lookup
    6. Klein O, Roded A, Hirschberg K, Fukuda M, Galli SJ, Sagi-Eisenberg R. Imaging FITC-dextran as a Reporter for Regulated Exocytosis.. J Vis Exp 2018 Jun 20;(136).
      doi: 10.3791/57936pubmed: 29985342google scholar: lookup
    7. Klein O, Roded A, Zur N, Azouz NP, Pasternak O, Hirschberg K, Hammel I, Roche PA, Yatsu A, Fukuda M, Galli SJ, Sagi-Eisenberg R. Rab5 is critical for SNAP23 regulated granule-granule fusion during compound exocytosis.. Sci Rep 2017 Nov 10;7(1):15315.
      doi: 10.1038/s41598-017-15047-8pubmed: 29127297google scholar: lookup
    8. Yuan T, Lu J, Zhang J, Zhang Y, Chen L. Spatiotemporal detection and analysis of exocytosis reveal fusion "hotspots" organized by the cytoskeleton in endocrine cells.. Biophys J 2015 Jan 20;108(2):251-60.
      doi: 10.1016/j.bpj.2014.11.3462pubmed: 25606674google scholar: lookup
    9. Kiskin NI, Babich V, Knipe L, Hannah MJ, Carter T. Differential cargo mobilisation within Weibel-Palade bodies after transient fusion with the plasma membrane.. PLoS One 2014;9(9):e108093.
      doi: 10.1371/journal.pone.0108093pubmed: 25233365google scholar: lookup
    10. Wu LG, Hamid E, Shin W, Chiang HC. Exocytosis and endocytosis: modes, functions, and coupling mechanisms.. Annu Rev Physiol 2014;76:301-31.
    11. Nightingale T, Cutler D. The secretion of von Willebrand factor from endothelial cells; an increasingly complicated story.. J Thromb Haemost 2013 Jun;11 Suppl 1(Suppl 1):192-201.
      doi: 10.1111/jth.12225pubmed: 23809123google scholar: lookup
    12. Cabeza JM, Acosta J, Alés E. Mechanisms of granule membrane recapture following exocytosis in intact mast cells.. J Biol Chem 2013 Jul 12;288(28):20293-305.
      doi: 10.1074/jbc.M113.459065pubmed: 23709219google scholar: lookup
    13. Kabaso D, Jorgačevski J, Calejo AI, Flašker A, Guček A, Kreft M, Zorec R. Comparison of unitary exocytic events in pituitary lactotrophs and in astrocytes: modeling the discrete open fusion-pore states.. Front Cell Neurosci 2013;7:33.
      doi: 10.3389/fncel.2013.00033pubmed: 23576951google scholar: lookup
    14. Lindau M. High resolution electrophysiological techniques for the study of calcium-activated exocytosis.. Biochim Biophys Acta 2012 Aug;1820(8):1234-42.
      doi: 10.1016/j.bbagen.2011.12.011pubmed: 22209782google scholar: lookup
    15. He L, Xue L, Xu J, McNeil BD, Bai L, Melicoff E, Adachi R, Wu LG. Compound vesicle fusion increases quantal size and potentiates synaptic transmission.. Nature 2009 May 7;459(7243):93-7.
      doi: 10.1038/nature07860pubmed: 19279571google scholar: lookup
    16. Stie J, Jesaitis AJ. Preparation of secretory vesicle-free plasma membranes by isopycnic sucrose gradient fractionation of neutrophils purified by the gelatin method.. Cytotechnology 2004 Oct;46(2-3):109-22.
      doi: 10.1007/s10616-005-0300-6pubmed: 19003266google scholar: lookup
    17. Valero V, Nevian T, Ho D, Lindau M. Tethering forces of secretory granules measured with optical tweezers.. Biophys J 2008 Nov 15;95(10):4972-8.
      doi: 10.1529/biophysj.108.132670pubmed: 18689450google scholar: lookup
    18. Neef A, Khimich D, Pirih P, Riedel D, Wolf F, Moser T. Probing the mechanism of exocytosis at the hair cell ribbon synapse.. J Neurosci 2007 Nov 21;27(47):12933-44.
    19. Kishimoto T, Liu TT, Hatakeyama H, Nemoto T, Takahashi N, Kasai H. Sequential compound exocytosis of large dense-core vesicles in PC12 cells studied with TEPIQ (two-photon extracellular polar-tracer imaging-based quantification) analysis.. J Physiol 2005 Nov 1;568(Pt 3):905-15.
      doi: 10.1113/jphysiol.2005.094003pubmed: 16150797google scholar: lookup
    20. Allersma MW, Wang L, Axelrod D, Holz RW. Visualization of regulated exocytosis with a granule-membrane probe using total internal reflection microscopy.. Mol Biol Cell 2004 Oct;15(10):4658-68.
      doi: 10.1091/mbc.e04-02-0149pubmed: 15282339google scholar: lookup
    21. Takahashi N, Hatakeyama H, Okado H, Miwa A, Kishimoto T, Kojima T, Abe T, Kasai H. Sequential exocytosis of insulin granules is associated with redistribution of SNAP25.. J Cell Biol 2004 Apr 26;165(2):255-62.
      doi: 10.1083/jcb.200312033pubmed: 15117968google scholar: lookup
    22. Gong LW, Hafez I, Alvarez de Toledo G, Lindau M. Secretory vesicles membrane area is regulated in tandem with quantal size in chromaffin cells.. J Neurosci 2003 Aug 27;23(21):7917-21.
    23. Zupancic G, Ogden D, Magnus CJ, Wheeler-Jones C, Carter TD. Differential exocytosis from human endothelial cells evoked by high intracellular Ca(2+) concentration.. J Physiol 2002 Nov 1;544(3):741-55.
      doi: 10.1113/jphysiol.2002.027490pubmed: 12411520google scholar: lookup
    24. Pothos EN, Mosharov E, Liu KP, Setlik W, Haburcak M, Baldini G, Gershon MD, Tamir H, Sulzer D. Stimulation-dependent regulation of the pH, volume and quantal size of bovine and rodent secretory vesicles.. J Physiol 2002 Jul 15;542(Pt 2):453-76.
      doi: 10.1113/jphysiol.2002.018630pubmed: 12122145google scholar: lookup
    25. Neuhaus EM, Almers W, Soldati T. Morphology and dynamics of the endocytic pathway in Dictyostelium discoideum.. Mol Biol Cell 2002 Apr;13(4):1390-407.
      doi: 10.1091/mbc.01-08-0392pubmed: 11950947google scholar: lookup
    26. Thompson RE, Lindau M, Webb WW. Robust, high-resolution, whole cell patch-clamp capacitance measurements using square wave stimulation.. Biophys J 2001 Aug;81(2):937-48.
      doi: 10.1016/S0006-3495(01)75752-9pubmed: 11463636google scholar: lookup
    27. Koh DS, Moody MW, Nguyen TD, Hille B. Regulation of exocytosis by protein kinases and Ca(2+) in pancreatic duct epithelial cells.. J Gen Physiol 2000 Oct;116(4):507-20.
      doi: 10.1085/jgp.116.4.507pubmed: 11004201google scholar: lookup
    28. Cochilla AJ, Angleson JK, Betz WJ. Differential regulation of granule-to-granule and granule-to-plasma membrane fusion during secretion from rat pituitary lactotrophs.. J Cell Biol 2000 Aug 21;150(4):839-48.
      doi: 10.1083/jcb.150.4.839pubmed: 10953007google scholar: lookup
    29. Williams RM, Shear JB, Zipfel WR, Maiti S, Webb WW. Mucosal mast cell secretion processes imaged using three-photon microscopy of 5-hydroxytryptamine autofluorescence.. Biophys J 1999 Apr;76(4):1835-46.
      doi: 10.1016/S0006-3495(99)77343-1pubmed: 10096882google scholar: lookup
    30. Thiel G, Battey N. Exocytosis in plants.. Plant Mol Biol 1998 Sep;38(1-2):111-25.
      pubmed: 9738963
    31. Scepek S, Coorssen JR, Lindau M. Fusion pore expansion in horse eosinophils is modulated by Ca2+ and protein kinase C via distinct mechanisms.. EMBO J 1998 Aug 3;17(15):4340-5.
      doi: 10.1093/emboj/17.15.4340pubmed: 9687502google scholar: lookup
    32. Lollike K, Borregaard N, Lindau M. Capacitance flickers and pseudoflickers of small granules, measured in the cell-attached configuration.. Biophys J 1998 Jul;75(1):53-9.
      doi: 10.1016/S0006-3495(98)77494-6pubmed: 9649367google scholar: lookup
    33. Oberhauser AF, Robinson IM, Fernandez JM. Simultaneous capacitance and amperometric measurements of exocytosis: a comparison.. Biophys J 1996 Aug;71(2):1131-9.
      doi: 10.1016/S0006-3495(96)79315-3pubmed: 8842250google scholar: lookup
    34. Hartmann J, Scepek S, Lindau M. Regulation of granule size in human and horse eosinophils by number of fusion events among unit granules.. J Physiol 1995 Feb 15;483 ( Pt 1)(Pt 1):201-9.
      doi: 10.1113/jphysiol.1995.sp020578pubmed: 7776232google scholar: lookup
    35. Lollike K, Borregaard N, Lindau M. The exocytotic fusion pore of small granules has a conductance similar to an ion channel.. J Cell Biol 1995 Apr;129(1):99-104.
      doi: 10.1083/jcb.129.1.99pubmed: 7535305google scholar: lookup