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Doxycycline reduction of f-actin content of human neutrophils and fibroblasts

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Abstract

We and others have shown that tetracyclines inhibit leukocyte adherence, migration, and phagocytosis, functions presumed to involve actin microfilament metabolism. In this study we investigated the influence of a tetracycline (doxycycline, Dc) on neutrophil and fibroblast actin metabolism. Human neutrophils and fibroblasts were pretreated with Dc or cytochalasin B (cB), stimulated with either the chemotactic peptide FMLP or phorbol myristate acetate (PMA), and the changes in phalloidin conjugate, associated F-actin were followed microscopically or quantified fluorometrically. Doxycycline suppressed, in a dose-related manner, the rise in F-actin content of neutrophils that normally follows their activation with either FMLP or PMA. Cytochalasin B had a similar affect on actin microfilament synthesis. Incubation of fibroblasts in Dc led to a loss of actin microfilaments and caused flattened adherent cells to round up and detach. Both Dc and cB also inhibited Con A-induced acceptor capping on neutrophils, a phenomenon known to be dependent on the presence of intact actin microfilaments. The data show that both Dc and cB influence actin metabolism and suggest they do so by differing mechanisms.

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References

  1. Bray, D., andWhite, J. G. 1988. Cortical flow in animal cells.Science 239:883.

    PubMed  Google Scholar 

  2. Omann, G. M., R. A. Allen, G. M. Bokoch, R. G. Painter, A. E. Traynor, andL. A. Sklar. 1987. Signal transduction and cytoskeletal activation in the neutrophil.Physiol. Rev. 67:285.

    PubMed  Google Scholar 

  3. White, J. R., P. H. Naccache, andR. I. Sha Afi. 1983. Stimulation by chemotactic factor of actin association with the cytoskeleton in rabbit neutrophils.J. Biol. Chem. 258:14041.

    PubMed  Google Scholar 

  4. Cassimeris, L., H. McNeill, andS. H. Zigmond. 1990. Chemoattractant-stimulated polymorphonuclear leukocytes contain two populations of actin filaments that differ in their spacial distribution and relative stabilities.J. Cell Biol. 110:1067.

    PubMed  Google Scholar 

  5. Gabler, W. L., and H. R.Creamer. Suppression of human neutrophil functions by tetracyclines.J. Periodont. Res. 26:52.

  6. Gabler, W. L., J. Smith, andN. Tsukuda. 1992. Comparison of doxycycline and a chemically modified tetracycline inhibition of leukocyte functions.Res. Commun. Chem. Pathol. Pharmacol. 78:151.

    PubMed  Google Scholar 

  7. Tsukada, N., and W. L.Gabler. 1993. The influence of doxycycline on the attachment of fibroblasts to gelatin coated surfaces and its cytotoxicity.J. Periodontol. (in press).

  8. Belsheim, J., C. Blomquist, J. Lofberg, andH. Gnarpe. 1983. Tetracycline influence on leukocyte functions.Acta Oto-Rhino-Laryngol. Belg. 37:635.

    Google Scholar 

  9. Gabler, W. L., W. W. Bullock, andH. R. Creamer. 1983. The endogenous component chemotactic assay (ECCA).Inflammation 7:321.

    PubMed  Google Scholar 

  10. Deguchi, S., S. Hori, H. R. Creamer, andW. L. Gabler. 1990. Neutrophil-mediated damage to human periodontal derived fibroblasts: Role of lipopolysaccharide.J. Periodont. Res. 25:293.

    PubMed  Google Scholar 

  11. Howard, T. H., andC. O. Oresajo. 1985. A method for quantifying F-actin in chemotactic peptide activated neutrophils: Study of the effect of fBOC peptide.Cell Motil. 5:545.

    PubMed  Google Scholar 

  12. Sklar, L. A., G. M. Omann, andR. G. Painter. 1985. Relationship of actin polymerization and depolymerization to light scattering in human neutrophils: Dependence on receptor occupancy and intracellular calcium.J. Cell Biol. 101:1161.

    PubMed  Google Scholar 

  13. Yuli, I., andR. Synderman. 1984. Light scattering by polymorphonuclear leukocytes exposed to chemoattractants. Discrete response correlate with chemotactic and secretory functions.J. Clin. Invest. 73:1408.

    PubMed  Google Scholar 

  14. May, S. W., N. Sahyoune, M. Wolf, andP. Custrecases. 1985. Role of intracellular calcium mobilization in the regulation of membrane processes.Nature 317:549.

    PubMed  Google Scholar 

  15. Svitkana, T. M., andA. A. Neyfaklr. 1986. Actin cytoskeleton of spread fibroblasts appears to assemble at the cell edge.J. Cell Sci. 82:235.

    PubMed  Google Scholar 

  16. Oliver, J. M., R. Lalchandani, andE. L. Becker. 1977. Actin redistribution during concanavalin A cap formation in rabbit neutrophils.J. Reticuloendothel. Soc. 21:359.

    PubMed  Google Scholar 

  17. Gabler, W. L. 1991. Fluxes and accumulation of tetracyclines in human blood cells.Res. Commun. Chem. Pathol. Pharmacol. 72:39.

    PubMed  Google Scholar 

  18. Ibsen, K. H., andM. R. Wist. 1964. The biochemistry and the physiology of the tetracyclines.Clin. Orthoped. Rel. Res. 32:143.

    Google Scholar 

  19. Pollard, T. D. 1990.Actin. Curr. Opin. Cell Biol. 2:33.

    Google Scholar 

  20. Van Meter, J. C., A. Spector, J. J. Oleson, andJ. H. Williams. 1952. In vitro action of aureomycin on oxidative phosphorylation in animal tissues.Proc. Soc. Exp. Biol. Med. 81:215.

    PubMed  Google Scholar 

  21. Loomis, W. F. 1950. On the action of aureomycin.Science 111:474.

    Google Scholar 

  22. Brody, T. M., R. Hurwitz, andJ. A. Bain. 1954. Magnesium and the effect of the tetracycline antibiotics on oxidative processes in mitochondria.Antibot. Chemother. 4:864.

    Google Scholar 

  23. Millonig, R., H. Salvo, andU. Aebi. 1986. Probing actin polymerization by intermolecular cross-linking.J. Cell Biol. 106:785.

    Google Scholar 

  24. Sklar, L. A., andZ. G. Oades. 1985. Signal transduction and ligand-receptor dynamics in the neutrophil. Ca2+ modulation and restoration.J. Biol. Chem. 260:11468.

    PubMed  Google Scholar 

  25. Pontremoli, S., E. Melloni, M. Michette, B. Sparatore, F. Salamino, O. Sacco, andB. L. Horecker. 1987. Phosphorylation and proteolytic modification of specific cytoskeletal proteins in human neutrophils stimulated by phorbol 12-myristate 12-acetate.Proc. Natl. Acad. Sci. U.S.A. 84:3604.

    PubMed  Google Scholar 

  26. Sheterline, P., J. E. Rickard, B. Boothroyd, andR. C. Richards. 1986. Phorbol ester induces rapid actin assembly in neutrophil leukocytes independently of changes in [Ca2+]i and pHi.J. Muscle Res. Cell Motil. 7:405.

    PubMed  Google Scholar 

  27. Gabler, W. L., W. W. Bullock, andH. R. Creamer. 1993. Phorbol myristate acetate induction of chemotactic migration of human polymorphonuclear leukocytes.Inflammation.17:521.

    PubMed  Google Scholar 

  28. Gabler, W. L. 1990. Potential roles of Mg2+ and Ca2+ in NADPH-oxidase dependent superoxide anion synthesis by human neutrophils.Res. Commun. Chem. Pathol. Pharmacol. 70:213.

    PubMed  Google Scholar 

  29. Yandekerchkhove, J. 1990. Actin-binding proteins.Curr. Opin. Cell Biol. 2 (No. 1):41.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Oral Molecular Biology, Oregon Health Sciences University, Portland, Oregon

    W. L. Gabler, J. Smith & N. Tsukuda

  2. Department of Biological Structure and Function, Oregon Health Sciences University, Portland, Oregon

    W. L. Gabler

  3. School of Dentistry, Oregon Health Sciences University, Portland, Oregon

    J. Smith

  4. Department of Periodontology, School of Dentistry, Hokkaido University, Sapporo, Japan

    N. Tsukuda

Authors
  1. W. L. Gabler
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  2. J. Smith
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  3. N. Tsukuda
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This research was supported by grants from the Kettering Family Foundation and the Medical Research Foundation of Oregon.

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Gabler, W.L., Smith, J. & Tsukuda, N. Doxycycline reduction of f-actin content of human neutrophils and fibroblasts. Inflammation 18, 107–118 (1994). https://doi.org/10.1007/BF01534603

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