Abstract
The effects of Con A on free cytoplasmic calcium concentrations in the cloned murine mast cell, MC9, have been measured using the fluorescent calcium indicator quin 2. Con A causes a rapid, small yet sustained rise in free cytosolic calcium (up to 245 nM) followed closely by increased45calcium uptake and more slowly by histamine release. The increases in45calcium uptake and histamine release require extracellular calcium. However, the Ca2+ influx blockers, nifedipine and verapamil inhibit these responses only at concentrations significantly higher than those used in smooth muscle to oppose potential-dependent events, and diltiazem is inactive. These observations suggest that, in these mast cells, other types of channels control Ca2+ entry.
In contrast, the intracellular Ca2+ blocker, TMB-8, inhibits both the Con A-induced histamine release and the Ca2+ changes. The calmodulin antagonists calmidazolium, trifluoperazine and W-7 are also highly effective inhibitors of both the Ca2+ changes and histamine release in direct proportion to their potency against calmodulin-dependent phosphodiesterase, implicating calmodulin in the regulation of stimulus-secretion in MC9 cells. These data imply that histamine release follows increases in intracellular Ca2+ concentration. Free intracellular Ca2+ results from rapid release from internal stores and is followed by a slower but more sustained influx of extracellular Ca2+.
Similar content being viewed by others
References
J. R. White, T. Ishizaka, K. Ishizaka and R. I. Sháafi,Direct demonstration of increased intracellular concentration of free calcium as measured by quin 2 in stimulated rat peritoneal mast cells. Proc. Nat. Acad. Sci. U.S.A.81, 3978–3982 (1984).
T. J. Rink, A. Sanchez and T. J. Hallam,Diacylglycerol and phorbol ester stimulate secretion without raising chytosolic free calcium in human platelet. Nature305, 317–319 (1983).
J. A. Ware, P. C. Johnson, M. Smith and E. W. Salzman,Aequorin detects increased cytoplasmic calcium in platelets stimulated with phorbol ester or diacylglycerol. Biochem. Biophys. Res. Comm.133, 98–104 (1985).
M. W. Musch, R. W. Bryant, C. Coscolluela, R. F. Myers and M. I. Siegel,Ionophore-stimulated lipoxygenase activity and histamine release in a cloned murine mast cells, MC9. Prostaglandins29, 405–430 (1985).
D. Guy-Grand, C. Griscelli and P. Vassali,The mouse gut T lymphocyte. J. Exp. Med.148, 1661–1667 (1978).
R. Y. Tsien, T. Pozzan and T. J. Rink,Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellular trapped fluorescent indicator. J. Cell Biol.84, 325–334 (1982).
R. Siraganian,An automated continuous-flow system for the extraction and fluorometric analysis of histamine. Anal. Biochem.57, 383–394 (1974).
N. S. Ranadive and N. D. Lanani,Movement of calcium ions and release of histamine from rat mast cells. Int. Archs. Allergy Appl. Immun.61, 9–18 (1980).
F. L. Pearce and J. R. White,Calcium efflux and histamine secretion from rat peritoneal mast cells. Agents and Actions14, 392–396 (1984).
R. Pervin, B. I. Kenner, G. Marx and E. Razin,Thrombin-induced degranulation of cultured bone marrow-derived mast cells: effect on calcium uptake. Immunology56, 667–672 (1985).
Y. Tanzaki and R. G. Townley,Effect of BSA on Ca 2+ influx in mast cells stimulated by ovalbumin. Int. Arch. Allergy Appl. Immun.70, 143–145 (1983).
E. Wolde Mussie and N. C. Moran,Histamine release by compound 48/80: evidence for the depletion and repletion of calcium using chlortetracycline and 45 calcium. Agents and Actions15, 267–272 (1984).
R. Y. Tsien, T. Pozzan and T. J. Rink,T cell mitogens cause early changes in cytosolic free Ca 2+ and membrane potential in lymphocytes. Nature295, 68–71 (1982).
T. J. Rink and T. Pozzan,Using quin 2 in cell suspensions. Cell Calcium6, 133–144 (1985).
S. J. Galli, A. M. Dvorak, J. A. Marcom, T. Ishizaka, G. Nabel, H. der Simonian, K. Pyne, J. M. Goldin, R. D. Rosenberg, H. Cantor and H. F. Dvorak,Mast cell clones: a model for the analysis of cellular maturation. J. Cell Biol.95, 435–444 (1982).
A. M. Magro and H. Bennick,Concanavalin A induced histamine release from human basophils in vitro. Immunology33, 51–58 (1977).
M. W. Musch and M. I. Siegel,Antigenic stimulated release of arachidonic acid, lipoxygenase activity and histamine release in a cloned murine mast cell, MC9. Biochem. Biophys. Research Comm.126, 512–525 (1985).
F. L. Pearce, M. Ennis, A. Truneh and J. R. White,Role of intra- and extracellular calcium in histamine release of rat peritoneal mast cells. Agents and Actions11, 51–54 (1981).
J. H. Baxter and R. Adamik,Differences in requirements and actions of various histamine releasing agents. Biochem. Pharmac.27, 497–503 (1978).
M. Ennis, A. Truneh, J. R. White and F. L. Pearce,Calcium pools involved in histamine release from rat mast cells. Int. Archs. Allergy Appl. Immun.62, 467–471 (1980).
A. J. Shores and J. L. Mongar,Modulation of histamine secretion from concanavalin A-activated rat mast cells by phosphatidylserine, calcium cAMP, pH and metabolic inhibitors. Agents and Actions10, 131–137 (1980).
M. Lindau and J. M. Fernandez,IgE-mediated degranulation of mast cells does not require opening of ion channels. Nature319, 150–153 (1986).
M. A. Beaven, J. Roger, J. P. Moore, T. R. Hesketh, G. A. Smith and M. C. Metcalf,The mechanism of the calcium signal and correlation with histamine release in 2H3 cells. J. Biol. Chem.259, 7129–7136 (1984).
R. A. Janis and D. J. Triggle,New developments in calcium channel antagonists. J. Med. Chem.26, 772–785 (1983).
D. J. Triggle,Calcium antagonists: basic chemical and pharmacological aspects, InNew perspectives on calcium antagonists, pp. 1–18 (Ed. G. B. Weiss). American Physiological Society, Bethesda 1981.
M. Ennis, P. W. Ind, F. L. Pearce and C. T. Dollery,Calcium antagonist and histamine secretion from rat peritoneal mast cells. Agent and Actions13, 144–148 (1983).
J. Sierchio, D. Ritchie, C. Bishop and R. Capetola,Calcium channel blockers in immediate hypersensitivity. Pharmacologist25, 120–124 (1983).
N. Chand, W. Diamantis, J. Pillar and R. D. Sofia,Inhibition of allergic and non-allergic histamine secretion from rat peritoneal mast cells by calcium antagonists. Br. J. Pharmac.83, 899–902 (1984).
C. Y. Chiow and M. H. Malagodi,Studies on the mechanism of action of a new calcium antagonist, TMB-8 hydrochloride in smooth and skeletal muscle. Br. J. Pharmac.53, 279–285 (1975).
N. Chakravarty and W. Yu,Role of intracellular calcium in histamine release from mast cells. Agents and Action14, 386–391 (1984).
N. Grossman,Effect of TMB-8 on histamine release from isolated rat mast cells. Int. Archs. Allergy Appl. Immun.79, 253–258 (1986).
H. Van Belle,The effect of drugs on calmodulin and its interaction with phosphodiesterase. Advances in Cyclic Nucleotide and protein Phosphorylation17, 557–567 (1984).
P. T. Peachell and F. L. Pearce,Effect of calmodulin inhibition on histamine secretion from mast cells. Agents and Actions16, 43–44 (1985).
T. Suzuki, K. Ohishi and M. Uchida,Effects of calmodulin inhibitors on histamine release from rat peritoneal mast cells induced by concanavalin A and ionophore A 23187. Gen. Pharm.14, 273–275 (1983).
B. Nilius, P. Hess, J. B. Landsman and R. W. Tsien,A novel type of cardiac calcium channel in ventricular cells. Nature316, 443–446 (1985).
H. A. Pershadsingh, R. Dane Gale, D. M. Delfert and J. M. McDonald,A calmodulin dependent Ca 2+-activated K + channel in the adipocyte plasma membrane. Biochem. Biophys. Research Comm.135, 934–941 (1986).
M. Donowitz,Ca 2+ in the control of active intestinal Na and Cl transport: involvement in neurohumoral action. Amer. J. Physiol.245 G165–177 (1983).
M. Donowitz, M. E. Cohen, R. Gudewich, L. Faylor and G. W. Sharp,Ca 2+-calmodulin, cyclic AMP and cyclic GMP-induced phosphorylation of proteins in purified microvillus membranes of rabbit ileum. Biochem. J.219, 573–581 (1984).
B. M. Curtis and W. A. Catterall,Phosphorylation of the calcium antagonist receptor of the voltage sensitive calcium channel by CAMP-dependent protein kinase. Proc. Natl. Acad. Sci. USA82, 2528–2532 (1985).
M. J. Berridge and R. F. Irvine,Inositoltrisphosphate, a novel second messenger in cellular signal transduction. Nature312, 315–321 (1984).
M. W. Musch and M. I. Siegel,Antigen-stimulated metabolism of inositol phospholipids in the cloned murine mast-cell line MC9. Biochem. J.234, 205–212 (1986).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Gulbenkian, A., Myers, J., Egan, R.W. et al. The role of a Ca2+/calmodulin dependent plasma membrane Ca2+ channel during Concanavalin A activation of MC9 mast cells. Agents and Actions 22, 16–23 (1987). https://doi.org/10.1007/BF01968811
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01968811