Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 Mar;85(6):1854–1858. doi: 10.1073/pnas.85.6.1854

Imaging asynchronous changes in intracellular Ca2+ in individual stimulated tumor mast cells.

P J Millard 1, D Gross 1, W W Webb 1, C Fewtrell 1
PMCID: PMC279879  PMID: 3162312

Abstract

Changes in the level of intracellular free calcium ([Ca2+]i) are associated with the secretion of mediators of immediate hypersensitivity by rat basophilic leukemia (RBL) cells, a mast cell line. Digital fluorescence ratio imaging of fura-2 was used to measure [Ca2+]i in individual RBL cells. Changes in [Ca2+]i that occurred in response to crosslinking of IgE receptors on the cell surface or to application of the Ca2+ ionophore ionomycin were studied. Stimulation of RBL cells with antigen resulted in rapid increases in [Ca2+]i following lag times that varied widely from cell to cell. Simple averaging of the Ca2+ responses of many cells yielded a gradual response profile that closely resembled that of suspensions of cells measured in the fluorometer. The results show that single cells can respond much more rapidly to antigen than has previously been suggested by studies on populations of cells. The lag time between addition of antigen and initiation of the increase in [Ca2+]i varied considerably between cells in the same field of view. Both the rise time and the variability and average duration of the lag time increased with decreasing antigen concentration.

Full text

PDF
1854

Images in this article

1856Image
on p.1856
1856Image
on p.1856

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Almers W., Neher E. The Ca signal from fura-2 loaded mast cells depends strongly on the method of dye-loading. FEBS Lett. 1985 Nov 11;192(1):13–18. doi: 10.1016/0014-5793(85)80033-8. [DOI] [PubMed] [Google Scholar]
  2. Barsumian E. L., Isersky C., Petrino M. G., Siraganian R. P. IgE-induced histamine release from rat basophilic leukemia cell lines: isolation of releasing and nonreleasing clones. Eur J Immunol. 1981 Apr;11(4):317–323. doi: 10.1002/eji.1830110410. [DOI] [PubMed] [Google Scholar]
  3. Beaven M. A., Rogers J., Moore J. P., Hesketh T. R., Smith G. A., Metcalfe J. C. The mechanism of the calcium signal and correlation with histamine release in 2H3 cells. J Biol Chem. 1984 Jun 10;259(11):7129–7136. [PubMed] [Google Scholar]
  4. Grynkiewicz G., Poenie M., Tsien R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem. 1985 Mar 25;260(6):3440–3450. [PubMed] [Google Scholar]
  5. Kruskal B. A., Keith C. H., Maxfield F. R. Thyrotropin-releasing hormone-induced changes in intracellular [Ca2+] measured by microspectrofluorometry on individual quin2-loaded cells. J Cell Biol. 1984 Sep;99(3):1167–1172. doi: 10.1083/jcb.99.3.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Liu F. T., Bohn J. W., Ferry E. L., Yamamoto H., Molinaro C. A., Sherman L. A., Klinman N. R., Katz D. H. Monoclonal dinitrophenyl-specific murine IgE antibody: preparation, isolation, and characterization. J Immunol. 1980 Jun;124(6):2728–2737. [PubMed] [Google Scholar]
  7. Mohr F. C., Fewtrell C. Depolarization of rat basophilic leukemia cells inhibits calcium uptake and exocytosis. J Cell Biol. 1987 Mar;104(3):783–792. doi: 10.1083/jcb.104.3.783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Neher E., Almers W. Fast calcium transients in rat peritoneal mast cells are not sufficient to trigger exocytosis. EMBO J. 1986 Jan;5(1):51–53. doi: 10.1002/j.1460-2075.1986.tb04176.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Rogers J., Hesketh T. R., Smith G. A., Beaven M. A., Metcalfe J. C., Johnson P., Garland P. B. Intracellular pH and free calcium changes in single cells using quene 1 and quin 2 probes and fluorescence microscopy. FEBS Lett. 1983 Sep 5;161(1):21–27. doi: 10.1016/0014-5793(83)80722-4. [DOI] [PubMed] [Google Scholar]
  10. Sawyer D. W., Sullivan J. A., Mandell G. L. Intracellular free calcium localization in neutrophils during phagocytosis. Science. 1985 Nov 8;230(4726):663–666. doi: 10.1126/science.4048951. [DOI] [PubMed] [Google Scholar]
  11. Taurog J. D., Fewtrell C., Becker E. L. IgE mediated triggering of rat basophil leukemia cells: lack of evidence for serine esterase activation. J Immunol. 1979 Jun;122(6):2150–2153. [PubMed] [Google Scholar]
  12. Tsien R. Y., Pozzan T., Rink T. J. Calcium homeostasis in intact lymphocytes: cytoplasmic free calcium monitored with a new, intracellularly trapped fluorescent indicator. J Cell Biol. 1982 Aug;94(2):325–334. doi: 10.1083/jcb.94.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Williams D. A., Fogarty K. E., Tsien R. Y., Fay F. S. Calcium gradients in single smooth muscle cells revealed by the digital imaging microscope using Fura-2. Nature. 1985 Dec 12;318(6046):558–561. doi: 10.1038/318558a0. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES