Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1991 Oct 1;115(1):59–66. doi: 10.1083/jcb.115.1.59

Calcium transients during Fc receptor-mediated and nonspecific phagocytosis by murine peritoneal macrophages

PMCID: PMC2289912  PMID: 1918139

Abstract

Studies with populations of macrophages have produced conflicting results concerning the possibility that the concentration of intracellular ionized calcium [( Ca2+]i) may act as an important mediator for phagocytosis. Since asynchronous changes in [Ca2+]i in individual cells undergoing phagocytosis may be averaged to undetectability in population studies, we studied single adhering murine macrophages using fura-2 and our previously described digital imaging system. The proportion of macrophages phagocytosing IgG-coated latex beads was greater than for uncoated beads (percent phagocytosing cells: 71 +/- 7 vs. 27 +/- 7, P less than 0.01). Phagocytosis of IgG- coated and uncoated beads was always associated with a calcium transient that preceded the initiation of phagocytosis. No calcium transients were detected in cells that bound but did not phagocytose beads. Four major differences between Fc receptor-mediated and nonspecific phagocytosis were detected: (a) the duration of calcium transients was longer for nonspecific phagocytosis compared with Fc receptor-mediated phagocytosis (69.9 +/- 10.2 vs. 48.7 +/- 4.7 s, P less than 0.05) and the magnitude of calcium transients was less for nonspecific phagocytosis (178 +/- 43 vs. 349 +/- 53 nM, P less than 0.05); (b) removal of extracellular calcium abolished the calcium transients associated with nonspecific phagocytosis but had no effect on those associated with receptor-mediated phagocytosis; (c) in the absence of extracellular calcium, buffering intracellular calcium with a chelator reduced Fc receptor-mediated phagocytosis but had no additive inhibitory effect on nonspecific phagocytosis; and (d) inhibition of protein kinase C (PKC) with staurosporine inhibited nonspecific phagocytosis but had no effect on receptor-mediated phagocytosis. Our observations suggest that despite both types of phagocytosis being associated with intracellular calcium transients, the role played by intracellular calcium in the signaling pathways may differ for Fc receptor-mediated and nonspecific phagocytosis by elicited murine macrophages.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. Albert P. R., Wolfson G., Tashjian A. H., Jr Diacylglycerol increases cytosolic free Ca2+ concentration in rat pituitary cells. Relationship to thyrotropin-releasing hormone action. J Biol Chem. 1987 May 15;262(14):6577–6581. [PubMed] [Google Scholar]
  2. Berridge M. J. Inositol trisphosphate and diacylglycerol: two interacting second messengers. Annu Rev Biochem. 1987;56:159–193. doi: 10.1146/annurev.bi.56.070187.001111. [DOI] [PubMed] [Google Scholar]
  3. Berridge M. J., Irvine R. F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. doi: 10.1038/312315a0. [DOI] [PubMed] [Google Scholar]
  4. Cheung J. Y., Constantine J. M., Bonventre J. V. Regulation of cytosolic free calcium concentration in cultured renal epithelial cells. Am J Physiol. 1986 Oct;251(4 Pt 2):F690–F701. doi: 10.1152/ajprenal.1986.251.4.F690. [DOI] [PubMed] [Google Scholar]
  5. Cheung J. Y., Tillotson D. L., Yelamarty R. V., Scaduto R. C., Jr Cytosolic free calcium concentration in individual cardiac myocytes in primary culture. Am J Physiol. 1989 Jun;256(6 Pt 1):C1120–C1130. doi: 10.1152/ajpcell.1989.256.6.C1120. [DOI] [PubMed] [Google Scholar]
  6. Cobbold P. H., Rink T. J. Fluorescence and bioluminescence measurement of cytoplasmic free calcium. Biochem J. 1987 Dec 1;248(2):313–328. doi: 10.1042/bj2480313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dedman J. R., Brinkley B. R., Means A. R. Regulation of microfilaments and microtubules by calcium and cyclic AMP. Adv Cyclic Nucleotide Res. 1979;11:131–174. [PubMed] [Google Scholar]
  8. Di Virgilio F., Meyer B. C., Greenberg S., Silverstein S. C. Fc receptor-mediated phagocytosis occurs in macrophages at exceedingly low cytosolic Ca2+ levels. J Cell Biol. 1988 Mar;106(3):657–666. doi: 10.1083/jcb.106.3.657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Hed J., Stendahl O. Differences in the ingestion mechanisms of IgG and C3b particles in phagocytosis by neutrophils. Immunology. 1982 Apr;45(4):727–736. [PMC free article] [PubMed] [Google Scholar]
  11. Kruskal B. A., Maxfield F. R. Cytosolic free calcium increases before and oscillates during frustrated phagocytosis in macrophages. J Cell Biol. 1987 Dec;105(6 Pt 1):2685–2693. doi: 10.1083/jcb.105.6.2685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lew D. P., Andersson T., Hed J., Di Virgilio F., Pozzan T., Stendahl O. Ca2+-dependent and Ca2+-independent phagocytosis in human neutrophils. Nature. 1985 Jun 6;315(6019):509–511. doi: 10.1038/315509a0. [DOI] [PubMed] [Google Scholar]
  13. McNeil P. L., Swanson J. A., Wright S. D., Silverstein S. C., Taylor D. L. Fc-receptor-mediated phagocytosis occurs in macrophages without an increase in average [Ca++]i. J Cell Biol. 1986 May;102(5):1586–1592. doi: 10.1083/jcb.102.5.1586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Miller B. A., Cheung J. Y., Tillotson D. L., Hope S. M., Scaduto R. C., Jr Erythropoietin stimulates a rise in intracellular-free calcium concentration in single BFU-E derived erythroblasts at specific stages of differentiation. Blood. 1989 Apr;73(5):1188–1194. [PubMed] [Google Scholar]
  15. Murphy E., Coll K., Rich T. L., Williamson J. R. Hormonal effects on calcium homeostasis in isolated hepatocytes. J Biol Chem. 1980 Jul 25;255(14):6600–6608. [PubMed] [Google Scholar]
  16. Nakadate T., Jeng A. Y., Blumberg P. M. Comparison of protein kinase C functional assays to clarify mechanisms of inhibitor action. Biochem Pharmacol. 1988 Apr 15;37(8):1541–1545. doi: 10.1016/0006-2952(88)90016-0. [DOI] [PubMed] [Google Scholar]
  17. Pollard T. D., Cooper J. A. Actin and actin-binding proteins. A critical evaluation of mechanisms and functions. Annu Rev Biochem. 1986;55:987–1035. doi: 10.1146/annurev.bi.55.070186.005011. [DOI] [PubMed] [Google Scholar]
  18. Scanlon M., Williams D. A., Fay F. S. A Ca2+-insensitive form of fura-2 associated with polymorphonuclear leukocytes. Assessment and accurate Ca2+ measurement. J Biol Chem. 1987 May 5;262(13):6308–6312. [PubMed] [Google Scholar]
  19. Silverstein S. C., Steinman R. M., Cohn Z. A. Endocytosis. Annu Rev Biochem. 1977;46:669–722. doi: 10.1146/annurev.bi.46.070177.003321. [DOI] [PubMed] [Google Scholar]
  20. Steinberg T. H., Newman A. S., Swanson J. A., Silverstein S. C. ATP4- permeabilizes the plasma membrane of mouse macrophages to fluorescent dyes. J Biol Chem. 1987 Jun 25;262(18):8884–8888. [PubMed] [Google Scholar]
  21. Stossel T. P. From signal to pseudopod. How cells control cytoplasmic actin assembly. J Biol Chem. 1989 Nov 5;264(31):18261–18264. [PubMed] [Google Scholar]
  22. Tamaoki T., Nomoto H., Takahashi I., Kato Y., Morimoto M., Tomita F. Staurosporine, a potent inhibitor of phospholipid/Ca++dependent protein kinase. Biochem Biophys Res Commun. 1986 Mar 13;135(2):397–402. doi: 10.1016/0006-291x(86)90008-2. [DOI] [PubMed] [Google Scholar]
  23. Yamaguchi D. T., Kleeman C. R., Muallem S. Protein kinase C-activated calcium channel in the osteoblast-like clonal osteosarcoma cell line UMR-106. J Biol Chem. 1987 Nov 5;262(31):14967–14973. [PubMed] [Google Scholar]
  24. Yelamarty R. V., Miller B. A., Scaduto R. C., Jr, Yu F. T., Tillotson D. L., Cheung J. Y. Three-dimensional intracellular calcium gradients in single human burst-forming units-erythroid-derived erythroblasts induced by erythropoietin. J Clin Invest. 1990 Jun;85(6):1799–1809. doi: 10.1172/JCI114638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Young J. D., Ko S. S., Cohn Z. A. The increase in intracellular free calcium associated with IgG gamma 2b/gamma 1 Fc receptor-ligand interactions: role in phagocytosis. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5430–5434. doi: 10.1073/pnas.81.17.5430. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES