Agents and Actions

, Volume 21, Issue 1–2, pp 62–65 | Cite as

Hyposmotic swelling leads to the expression of trypsin-like activity by rat peritoneal fluid mast cells

  • C. S. Vassimon
  • E. T. Gomes
  • A. M. Rothschild
Histamine and Kinins


Rat peritoneal fluid mast cell present parallel increases in cell area (swelling), and in hydrolytic activity on the trypsin substrate p-tosyl arginine methyl ester (TAME) when placed in Tris buffers of concentrations between 0.15 and 0.03M. Under these conditions, cells do not degranulate and preserve their trypsin-like enzyme activity after low speed centrifugation. Exposure to more dilute Tris buffers, between 0.015 and 0.003M, leads to cell rupture accompanied by progressive degranulation and loss of activity on TAME. Protamine, a heparin antagonist prevented this loss when added to mast cells prior to hyposmotic lysis, or lysis by sonication or repeated periods of freezing and thawing. Enzyme activity released in the presence of protamine was fully recovered in supernates of cell lysates submitted to low speed centrifugation. Controlled swelling of mast cells propitiates the expression of trypsin-like activity, possibly by facilitating enzyme-substrate interaction. Cell lysis on the contrary, leads to inactivation of such activity, possibly by enzyme binding to heparin in exposed mast cell granules.


Heparin Mast Cell Methyl Ester Cell Granule Tris Buffer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    T. Ishizaka,Analysis of triggering events in mast cells for immunoglobulin E-mediated histamine release. J. Aller. Clin. Immunol.67, 90–96 (1981).Google Scholar
  2. [2]
    G. G. Glenner, V. K. Hopsu and L. A. Cohen,Histochemical demonstration of trypsin-like esterase activity in mast cells. J. Histochem. Cytochem.10, 109–114 (1962).Google Scholar
  3. [3]
    Y. Katayama, J. V. Auditore and N. Endo,Purification and characterization of a mouse mast cell esteroprotease. Exp. Med. Pathol.14, 228–235 (1971).Google Scholar
  4. [4]
    B. Schwartz, R. A. Lewis and K. F. Austen,Tryptase from human pulmonary mast cells. J. Biol. Chem.256, 11939–11949 (1981).Google Scholar
  5. [5]
    A. M. Rothschild,Hydrolysis of arginine and tyrosine esters in mast cells exposed to epinephrine. Biochem. Pharmac.29, 419–427 (1980).Google Scholar
  6. [6]
    H. Kido, N. Fukusen and N. Katunuma,Chymotrypsin and trypsin type serine proteases in rat mast cells: Properties and functions. Arch. Biochem. Biophys.239, 436–443 (1985).Google Scholar
  7. [7]
    D. Lagunoff and E. P. Benditt,Proteolytic enzymes of mast cells. Ann. N.Y. Acad. Sci.103, 185–198 (1963).Google Scholar
  8. [8]
    J. Kawiak, W. H. Vensel, J. Komender and E. A. Barnard,Non-pancreatic proteases of the chymotrypsin family. I. A. Chymotrypsin-like protease from rat mast cell. Biochem. Biophys. Acta235, 172–181 (1971).Google Scholar
  9. [9]
    S. Okuno-Kaneda, T. Saito, T. Kawasaki, A. Ichikawa and K. Tomita,Properties of protease in mast cell granules. Biochem. Pharmac.29, 1715–1722 (1980).Google Scholar
  10. [10]
    Z. Darzynkewicz and E. A. Barnard,Specific proteases of the rat mast cell. Nature213, 1198–1204 (1967).Google Scholar
  11. [11]
    E. L. Tamburus and A. M. Rothschild,Reversible swelling evoked by epinephrine, 8-bromo cyclic 3′5′-guanosine monophosphate and feeding in the mast cells of the rat. Biochem. Pharmac.30, 469–479 (1981).Google Scholar
  12. [12]
    S. Grinstein, A. Rothstein, A. Sarkadi and E. W. Gelfand,Responses of lymphocytes to anisotonic media: volume regulating behavior. Am. J. Physiol.246, c204-c215 (1984).Google Scholar
  13. [13]
    G. D. Bloom and O. Haegermark,Studies on morphological changes and histamine release induced by bee venom, n-decylamine and hypotonic solutions in rat peritoneal mast cells. Acta. Physiol. Scan.71, 257–269 (1967).Google Scholar
  14. [14]
    B. C. W. Hummel,A modified spectrophotometric determination of chymotrypsin, trypsin, and thrombin. Can. J. Biochem. Physiol.37, 1393–1398 (1959).Google Scholar
  15. [15]
    L. Enebrack and I. Svensson,Isolation of rat peritoneal mast cells by centrifugation on density gradients of Percoll. J. Immunol. Meth.39, 135–145 (1980).Google Scholar
  16. [16]
    L. B. Schwartz, C. Riedel, J. P. Caulfield, S. I. Wasserman and K. F. Austen,Cell association of complexes of chymase, heparin proteoglycan and protein after degranulation by rat mast cells. J. Immunol.126, 2071–2078 (1981).Google Scholar
  17. [17]
    L. B. Schwartz, C. Riedel, J. J. Schratz and K. F. Austen,Localization of carboxypeptidase A to the macromolecular heparin proteoglycan-protein complex in secretory granules of rat serosal mast cells. J. Immunol.128, 1128–1133 (1982).Google Scholar
  18. [18]
    B. Uvnäs,The role of the heparin protein complex in the storage of histamine in mast cells. InStructure, Cellular Functions and Clinical Applications (Ed. N. M. McDuffie), Academic Press, New York (1979).Google Scholar
  19. [19]
    G. D. Bloom and N. Chakravarty,Time course of anaphylatic histamine release and morphological changes in rat peritoneal mast cells. Acta. Physiol. Scand.78, 410–416 (1970).Google Scholar
  20. [20]
    P. Anderson, S. A. Slorach and B. Uvnäs,Sequential exocytosis of storage granules during antigen-induced histmaine release form sensitized and mast cells in vitro. An electron microscopic study. Acta. Physiol. Scand.88, 359–365 (1973).Google Scholar
  21. [21]
    M. P. Oliveira and A. M. Rothschild,Effects of catecholamines on rat mesentery mast cells. Nature218, 382 (1968).Google Scholar
  22. [22]
    A. M. Rothschild,Plasma kallikrein-generating activity evoked by rat peritoneal fluid mast cells following treatment with epinephrine, 8-bromo-cyclic 3′5′-guanosine monophosphate or compound 48/80. Biochem. Pharmacol.30, 481–487 (1981).Google Scholar
  23. [23]
    A. M. Rothschild and E. L. T. Gomes,Stimulation by epinephrine of histamine synthesis by rat peritoneal fluid mast cells in vitro. Life Sci.35, 1645–1652 (1984).Google Scholar

Copyright information

© Birkhäuser Verlag 1987

Authors and Affiliations

  • C. S. Vassimon
    • 1
  • E. T. Gomes
    • 1
  • A. M. Rothschild
    • 1
  1. 1.Department of Pharmacology, School of Medicine of Ribeiräo PretoUniversity of São PauloRibeirão PretoBrasil

Personalised recommendations