Abstract
When macrophages and neutrophils are allowed to settle onto an appropriate surface, they attach and spread in a frustrated attempt to phagocytose the substrate. Spreading is associated with extensive rearrangements of the actin cytoskeleton which resemble those occurring during phagocytosis. We have previously shown that spreading in human neutrophils is preceded by an increase in cytosolic-free calcium concentration [( Ca2+]i) (Kruskal, B. A., S. Shak, and F. R. Maxfield. 1986. Proc. Natl. Acad. Sci. USA. 83:2919-2923). To assess the generality of this signal, we measured [Ca2+]i in single thioglycollate- elicited mouse peritoneal macrophages as they spread on an immune complex-coated surface, using fura-2 microspectrofluorometry. A [Ca2+]i increase always precedes spreading. This increase can involve several (up to 8) [Ca2+]i spikes, with an average peak value of 387 +/- 227 nM (mean +/- SD, n = 92 peaks in 24 cells), before spreading is detected. Neither spreading nor the magnitude of these spikes is significantly altered by removal of extracellular calcium. Many of the spreading macrophages exhibit periodic [Ca2+]i increases before and during spreading. The proportion which does so varies among experiments from 0 to 90%, but it is frequently greater than 40%. The largest number of cells (approximately 25%) exhibited only a single peak. In 13 cells that showed more than 10 peaks, the median period was 29 s (range 19-69 s). The average peak [Ca2+]i was 385 +/- 266 nM (mean +/- SD, n = 208 peaks in 14 cells). The calcium producing these increases is derived from intracellular pools. The oscillations occur with spreading on either opsonized or nonopsonized surfaces. The function of these oscillations is not clear, but the large number of cells which exhibit them suggest that they may be important to macrophage function.
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Selected References
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- Boyles J., Bainton D. F. Changes in plasma-membrane-associated filaments during endocytosis and exocytosis in polymorphonuclear leukocytes. Cell. 1981 Jun;24(3):905–914. doi: 10.1016/0092-8674(81)90116-1. [DOI] [PubMed] [Google Scholar]
- Boyles J., Bainton D. F. Changing patterns of plasma membrane-associated filaments during the initial phases of polymorphonuclear leukocyte adherence. J Cell Biol. 1979 Aug;82(2):347–368. doi: 10.1083/jcb.82.2.347. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gallin E. K., Gallin J. I. Interaction of chemotactic factors with human macrophages. Induction of transmembrane potential changes. J Cell Biol. 1977 Oct;75(1):277–289. doi: 10.1083/jcb.75.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gallin E. K., Wiederhold M. L., Lipsky P. E., Rosenthal A. S. Spontaneous and induced membrane hyperpolarizations in macrophages. J Cell Physiol. 1975 Dec;86 (Suppl 2)(3 Pt 2):653–661. doi: 10.1002/jcp.1040860510. [DOI] [PubMed] [Google Scholar]
- 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]
- Ince C., Leijh P. C., Meijer J., Van Bavel E., Ypey D. L. Oscillatory hyperpolarizations and resting membrane potentials of mouse fibroblast and macrophage cell lines. J Physiol. 1984 Jul;352:625–635. doi: 10.1113/jphysiol.1984.sp015313. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ives H. E., Daniel T. O. Interrelationship between growth factor-induced pH changes and intracellular Ca2+. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1950–1954. doi: 10.1073/pnas.84.7.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kouri J., Noa M., Diaz B., Niubo E. Hyperpolarisation of rat peritoneal macrophages phagocytosing latex particles. Nature. 1980 Feb 28;283(5750):868–869. doi: 10.1038/283868a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Kruskal B. A., Shak S., Maxfield F. R. Spreading of human neutrophils is immediately preceded by a large increase in cytoplasmic free calcium. Proc Natl Acad Sci U S A. 1986 May;83(9):2919–2923. doi: 10.1073/pnas.83.9.2919. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Michl J., Pieczonka M. M., Unkeless J. C., Silverstein S. C. Effects of immobilized immune complexes on Fc- and complement-receptor function in resident and thioglycollate-elicited mouse peritoneal macrophages. J Exp Med. 1979 Sep 19;150(3):607–621. doi: 10.1084/jem.150.3.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nelson P. G., Henkart M. P. Oscillatory membrane potential changes in cells of mesenchymal origin: the role of an intracellular calcium regulating system. J Exp Biol. 1979 Aug;81:49–61. doi: 10.1242/jeb.81.1.49. [DOI] [PubMed] [Google Scholar]
- North R. J. The uptake of particulate antigens. J Reticuloendothel Soc. 1968 Jun;5(3):203–229. [PubMed] [Google Scholar]
- Okada Y., Doida Y., Roy G., Tsuchiya W., Inouye K., Inouye A. Oscillations of membrane potential in L cells. I. Basic characteristics. J Membr Biol. 1977 Aug 4;35(4):319–335. doi: 10.1007/BF01869957. [DOI] [PubMed] [Google Scholar]
- Okada Y., Tsuchiya W., Yada T. Calcium channel and calcium pump involved in oscillatory hyperpolarizing responses of L-strain mouse fibroblasts. J Physiol. 1982 Jun;327:449–461. doi: 10.1113/jphysiol.1982.sp014242. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Okada Y., Tsuchiya W., Yada T., Yano J., Yawo H. Phagocytic activity and hyperpolarizing responses in L-strain mouse fibroblasts. J Physiol. 1981;313:101–119. doi: 10.1113/jphysiol.1981.sp013653. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Poenie M., Alderton J., Steinhardt R., Tsien R. Calcium rises abruptly and briefly throughout the cell at the onset of anaphase. Science. 1986 Aug 22;233(4766):886–889. doi: 10.1126/science.3755550. [DOI] [PubMed] [Google Scholar]
- 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]
- Rabinovitch M., DeStefano M. J. Macrophage spreading in vitro. I. Inducers of spreading. Exp Cell Res. 1973 Mar 15;77(1):323–334. doi: 10.1016/0014-4827(73)90584-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Siffert W., Akkerman J. W. Activation of sodium-proton exchange is a prerequisite for Ca2+ mobilization in human platelets. 1987 Jan 29-Feb 4Nature. 325(6103):456–458. doi: 10.1038/325456a0. [DOI] [PubMed] [Google Scholar]
- Southwick F. S., Stossel T. P. Contractile proteins in leukocyte function. Semin Hematol. 1983 Oct;20(4):305–321. [PubMed] [Google Scholar]
- Trotter J. A. The organization of actin in spreading macrophages. The actin-cytoskeleton of peritoneal macrophages is linked to the substratum via transmembrane connections. Exp Cell Res. 1981 Apr;132(2):235–248. doi: 10.1016/0014-4827(81)90099-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Tsien R. Y., Rink T. J., Poenie M. Measurement of cytosolic free Ca2+ in individual small cells using fluorescence microscopy with dual excitation wavelengths. Cell Calcium. 1985 Apr;6(1-2):145–157. doi: 10.1016/0143-4160(85)90041-7. [DOI] [PubMed] [Google Scholar]
- Ueda S., Oiki S., Okada Y. Oscillations of cytoplasmic concentrations of Ca2+ and K+ in fused L cells. J Membr Biol. 1986;91(1):65–72. doi: 10.1007/BF01870215. [DOI] [PubMed] [Google Scholar]
- Watanabe S., Smith C. R., Phillips M. J. Coordination of the contractile activity of bile canaliculi. Evidence from calcium microinjection of triplet hepatocytes. Lab Invest. 1985 Sep;53(3):275–279. [PubMed] [Google Scholar]
- Woods N. M., Cuthbertson K. S., Cobbold P. H. Repetitive transient rises in cytoplasmic free calcium in hormone-stimulated hepatocytes. Nature. 1986 Feb 13;319(6054):600–602. doi: 10.1038/319600a0. [DOI] [PubMed] [Google Scholar]
- 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]