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. 1971 Nov 1;51(2):465–483. doi: 10.1083/jcb.51.2.465

ELECTRON MICROSCOPE OBSERVATIONS ON COMPOUND 48/80-INDUCED DEGRANULATION IN RAT MAST CELLS

Evidence for Sequential Exocytosis of Storage Granules

Pál Röhlich 1, Per Anderson 1, Börje Uvnäs 1
PMCID: PMC2108143  PMID: 4107023

Abstract

In vitro degranulation of rat mast cells was studied at different intervals ranging from 10 to 60 sec after adding the histamine liberator, compound 48/80 (0.4 µg/ml, 17°C). The ultrastructural changes were followed by electron microscopy, and parallel assays were made to determine the histamine released. In addition, the extracellular tracers lanthanum and hemoglobin (demonstrated by its peroxidative activity) were applied to mast cells to follow communication of the extracellular space with the cavities formed during degranulation. After a lag period of 10 sec, degranulation started in the most peripherally located granules. The perigranular membrane fused with the plasma membrane, resulting in a pore bridged by a thin diaphragm. This was followed by rupture of the diaphragm and extrusion of the granule matrix (exocytosis). The process advanced towards the cell interior by fusion and opening of the deeper situated granules to the formerly opened granule cavities. At the end of the process, the cell was filled by a system of complicated cavities containing a number of altered granules. Extracellular tracers have shown that these intracellular cavities were in unbroken communication with the extracellular space from the very beginning of their formation. Both lanthanum and hemoglobin were found to be adsorbed to the limiting membrane of the cavities and bound to altered mast cell granules. In contrast, no tracer substance was present in nondegranulating mast cells. Degranulation of mast cells by compound 48/80 is regarded as a sequential exocytosis, a process similar to that described for some exocrine gland cells. All the "intracellular" cavities, formed by degranulation, were shown to communicate with the extracellular space; consequently, granules lying in these cavities must be considered as biologically extracellular. The present findings support the view that histamine is released from the granule matrix by the extracellular ionic milieu.

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Selected References

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  1. Aborg C. H., Novotný J., Uvnäs B. Ionic binding of histamine in mast cell granules. Acta Physiol Scand. 1967 Apr;69(4):276–283. doi: 10.1111/j.1748-1716.1967.tb03523.x. [DOI] [PubMed] [Google Scholar]
  2. Amsterdam A., Ohad I., Schramm M. Dynamic changes in the ultrastructure of the acinar cell of the rat parotid gland during the secretory cycle. J Cell Biol. 1969 Jun;41(3):753–773. doi: 10.1083/jcb.41.3.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bloom G. D., Chakravarty N. Time course of anaphylactic histamine release and morphological changes in rat peritoneal mast cells. Acta Physiol Scand. 1970 Mar;78(3):410–419. doi: 10.1111/j.1748-1716.1970.tb04677.x. [DOI] [PubMed] [Google Scholar]
  4. Bloom G. D., Fredholm B., Haegermark O. Studies on the time course of histamine release and morphological changes induced by histamine liberators in rat peritoneal mast cells. Acta Physiol Scand. 1967 Dec;71(4):270–282. doi: 10.1111/j.1748-1716.1967.tb03734.x. [DOI] [PubMed] [Google Scholar]
  5. Bloom G. D., Haegermark O. A study on morphological changes and histamine release induced by compound 48/80 in rat peritoneal mast cells. Exp Cell Res. 1965 Dec;40(3):637–654. doi: 10.1016/0014-4827(65)90241-7. [DOI] [PubMed] [Google Scholar]
  6. Bloom G. D., Haegermark O. Studies on morphological changes and histamine release induced by bee venom, n-decylamine and hypotonic solutions in rat peritoneal mast cells. Acta Physiol Scand. 1967 Dec;71(4):257–269. doi: 10.1111/j.1748-1716.1967.tb03733.x. [DOI] [PubMed] [Google Scholar]
  7. Carlsson S. A., Ritzén M. Mast cells and 5-HT. Intracellular release of 5-hydroxytryptamine (5-HT) from storage granules during anaphylaxis or treatment with compound 48-80. Acta Physiol Scand. 1969 Dec;77(4):449–464. doi: 10.1111/j.1748-1716.1969.tb04589.x. [DOI] [PubMed] [Google Scholar]
  8. Chakravarty N., Gustafson G. T., Pihl E. Ultrastructural changes in rat mast cells during anaphylactic histamine release. Acta Pathol Microbiol Scand. 1967;71(2):233–244. doi: 10.1111/j.1699-0463.1967.tb05161.x. [DOI] [PubMed] [Google Scholar]
  9. Cohen A. I. New evidence supporting the linkage to extracellular space of outer segment saccules of frog cones but not rods. J Cell Biol. 1968 May;37(2):424–444. doi: 10.1083/jcb.37.2.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Combs J. W. Maturation of rat mast cells. An electron microscope study. J Cell Biol. 1966 Dec;31(3):563–575. doi: 10.1083/jcb.31.3.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cotran K. S., Karnovsky M. J. Vascular leakage induced by horseradish peroxidase in the rat. Proc Soc Exp Biol Med. 1967 Nov;126(2):557–561. doi: 10.3181/00379727-126-32504. [DOI] [PubMed] [Google Scholar]
  12. FAWCETT D. W. An experimental study of mast cell degranulation and regeneration. Anat Rec. 1955 Jan;121(1):29–51. doi: 10.1002/ar.1091210104. [DOI] [PubMed] [Google Scholar]
  13. Fillion G. M., Slorach S. A., Uvnäs B. The release of histamine, heparin and granule protein from rat mast cells treated with compound 48-80 in vitro. Acta Physiol Scand. 1970 Apr;78(4):547–560. doi: 10.1111/j.1748-1716.1970.tb04691.x. [DOI] [PubMed] [Google Scholar]
  14. Goldfischer S., Novikoff A. B., Albala A., Biempica L. Hemoglobin uptake by rat hepatocytes and its breakdown within lysosomes. J Cell Biol. 1970 Mar;44(3):513–529. doi: 10.1083/jcb.44.3.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  16. Hand A. R. The fine structure of von Ebner's gland of the rat. J Cell Biol. 1970 Feb;44(2):340–353. doi: 10.1083/jcb.44.2.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Horsfield G. I. The effect of compound 48/80 on the rat mast cell. J Pathol Bacteriol. 1965 Oct;90(2):599–605. doi: 10.1002/path.1700900228. [DOI] [PubMed] [Google Scholar]
  18. ICHIKAWA A. FINE STRUCTURAL CHANGES IN RESPONSE TO HORMONAL STIMULATION OF THE PERFUSED CANINE PANCREAS. J Cell Biol. 1965 Mar;24:369–385. doi: 10.1083/jcb.24.3.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Johnson A. R., Moran N. C. Release of histamine from rat mast cells: a comparison of the effects of 48-80 and two antigen systems. Fed Proc. 1969 Sep-Oct;28(5):1716–1720. [PubMed] [Google Scholar]
  20. Kluge T. The permeability of mesothelium to horseradish peroxidase. A light and electron microscopic study, with special reference to the morphology of pericardial mast cells. Acta Pathol Microbiol Scand. 1969;75(2):257–269. [PubMed] [Google Scholar]
  21. LAGUNOFF D., BENDITT E. P. Proteolytic enzymes of mast cells. Ann N Y Acad Sci. 1963 Feb 26;103:185–198. doi: 10.1111/j.1749-6632.1963.tb53698.x. [DOI] [PubMed] [Google Scholar]
  22. LAGUNOFF D., PHILLIPS M. T., ISERI O. A., BENDITT E. P. ISOLATION AND PRELIMINARY CHARACTERIZATION OF RAT MAST CELL GRANULES. Lab Invest. 1964 Nov;13:1331–1344. [PubMed] [Google Scholar]
  23. Mann P. R. An electron microscope study of the degranulation of rat peritoneal mast cells brought about by four different agents. Br J Dermatol. 1969 Dec;81(12):926–936. doi: 10.1111/j.1365-2133.1969.tb15975.x. [DOI] [PubMed] [Google Scholar]
  24. Nósal R., Slorach S. A., Uvnäs B. Quantitative correlation between degranulation and histamine release following exposure of rat mast cells to compound 48-80 in vitro. Acta Physiol Scand. 1970 Oct;80(2):215–221. doi: 10.1111/j.1748-1716.1970.tb04785.x. [DOI] [PubMed] [Google Scholar]
  25. Padawer J., Fruhman G. J. Phagocytosis of zymosan particles by mast cells. Experientia. 1968 May 15;24(5):471–472. doi: 10.1007/BF02144399. [DOI] [PubMed] [Google Scholar]
  26. Padawer J. Ingestion of colloidal gold by mast cells. Proc Soc Exp Biol Med. 1968 Dec;129(3):905–907. doi: 10.3181/00379727-129-33455. [DOI] [PubMed] [Google Scholar]
  27. Padawer J. Uptake of colloidal thorium dioxide by mast cells. J Cell Biol. 1969 Mar;40(3):747–760. doi: 10.1083/jcb.40.3.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. SHORE P. A., BURKHALTER A., COHN V. H., Jr A method for the fluorometric assay of histamine in tissues. J Pharmacol Exp Ther. 1959 Nov;127:182–186. [PubMed] [Google Scholar]
  29. Setoguti T. Electron microscopic study on the newt nmast cell, especially its granule-extrusion mechanism. J Ultrastruct Res. 1969 Jun;27(5):377–395. doi: 10.1016/s0022-5320(69)80038-9. [DOI] [PubMed] [Google Scholar]
  30. Singleton E. M., Clark S. L., Jr The response of mast cells to compound 48/80 studied with the electron microscope. Lab Invest. 1965 Oct;14(10):1744–1763. [PubMed] [Google Scholar]
  31. Thon I. L., Uvnäs B. Degranulation and histamine release, two consecutive steps in the response of rat mast cells to compound 48-80. Acta Physiol Scand. 1967 Dec;71(4):303–315. doi: 10.1111/j.1748-1716.1967.tb03737.x. [DOI] [PubMed] [Google Scholar]
  32. Thon I. L., Uvnäs B. Mode of storage of histamine in mast cells. Acta Physiol Scand. 1966 Jul-Aug;67(3):455–470. doi: 10.1111/j.1748-1716.1966.tb03332.x. [DOI] [PubMed] [Google Scholar]
  33. UVNAES B. RELEASE PROCESSES IN MAST CELLS AND THEIR ACTIVATION BY INJURY. Ann N Y Acad Sci. 1964 Aug 27;116:880–890. doi: 10.1111/j.1749-6632.1964.tb52554.x. [DOI] [PubMed] [Google Scholar]
  34. UVNAS B. Mechanism of action of a histamine-liberating principle in jellyfish (Cyanea capillata). Ann N Y Acad Sci. 1960 Nov 17;90:751–759. doi: 10.1111/j.1749-6632.1960.tb26419.x. [DOI] [PubMed] [Google Scholar]
  35. Uvnäs B., Wold J. K. Isolation of a mast cell degranulating polypeptide from Ascaris suis. Acta Physiol Scand. 1967 Jul-Aug;70(3):269–276. doi: 10.1111/j.1748-1716.1967.tb03625.x. [DOI] [PubMed] [Google Scholar]
  36. Wolff J., Merker H. J. Ultrastruktur und Bildung von Poren im Endothel von porösen und geschlossenen Kapillaren. Z Zellforsch Mikrosk Anat. 1966;73(2):174–191. [PubMed] [Google Scholar]

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