Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1980 Aug 1;86(2):634–640. doi: 10.1083/jcb.86.2.634

Phorbol myristate acetate stimulates pinocytosis and membrane spreading in mouse peritoneal macrophages

PMCID: PMC2111483  PMID: 7400219

Abstract

Phorbol myristate acetate (PMA) at a concentration of 0.01 microgram/ml causes an approximately threefold increase in surface area of resident, proteose-peptone-elicited, and thioglycolate-broth-elicited mouse peritoneal macrophages. Resident and proteose-peptone-elicited macrophages, cultured for 24 h in the presence of PMA, increase their pinocytic rate twofold in response to addition of PMA (0.01 microgram/ml) to the medium. Thioglycolate-broth-elicited macrophages, cultured for 24 h in the absence of PMA, immediately increase their pinocytic rate 2- to 3.5-fold in response to a single challenge with PMA (0.01 microgram/ml). Cytochalasin B, colchicine, and podophyllotoxin have only modest inhibitory effects on the basal rate of pinocytosis and on PMA-induced cellular spreading, but completely block the stimulatory effects of PMA on pinocytosis in thioglycolate- broth-elicited macrophages. Cytochalasin D markedly inhibits both basal and PMA-stimulated pinocytosis in these cells. Thus, PMA is a useful tool for studying mechanisms of macrophage spreading and for enhancing the overall rate of pinosome formation.

Full Text

The Full Text of this article is available as a PDF (886.1 KB).

Selected References

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

  1. Brune K., Glatt M., Kälin H., Peskar B. A. Pharmacological control of prostaglandin and thromboxane release from macrophages. Nature. 1978 Jul 20;274(5668):261–263. doi: 10.1038/274261a0. [DOI] [PubMed] [Google Scholar]
  2. Driedger P. E., Blumberg P. M. Specific binding of phorbol ester tumor promoters. Proc Natl Acad Sci U S A. 1980 Jan;77(1):567–571. doi: 10.1073/pnas.77.1.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Edelson P. J., Zwiebel R., Cohn Z. A. The pinocytic rate of activated macrophages. J Exp Med. 1975 Nov 1;142(5):1150–1164. doi: 10.1084/jem.142.5.1150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Estensen R. D., White J. G., Holmes B. Specific degranulation of human polymorphonuclear leukocytes. Nature. 1974 Mar 22;248(446):347–348. doi: 10.1038/248347a0. [DOI] [PubMed] [Google Scholar]
  5. Goldstein J. L., Brown M. S. The low-density lipoprotein pathway and its relation to atherosclerosis. Annu Rev Biochem. 1977;46:897–930. doi: 10.1146/annurev.bi.46.070177.004341. [DOI] [PubMed] [Google Scholar]
  6. Granelli-Piperno A., Vassalli J. D., Reich E. Secretion of plasminogen activator by human polymorphonuclear leukocytes. Modulation by glucocorticoids and other effectors. J Exp Med. 1977 Dec 1;146(6):1693–1706. doi: 10.1084/jem.146.6.1693. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Griffin F. M., Jr, Bianco C., Silverstein S. C. Characterization of the macrophage receptro for complement and demonstration of its functional independence from the receptor for the Fc portion of immunoglobulin G. J Exp Med. 1975 Jun 1;141(6):1269–1277. doi: 10.1084/jem.141.6.1269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Griffin F. M., Jr, Griffin J. A., Silverstein S. C. Studies on the mechanism of phagocytosis. II. The interaction of macrophages with anti-immunoglobulin IgG-coated bone marrow-derived lymphocytes. J Exp Med. 1976 Sep 1;144(3):788–809. doi: 10.1084/jem.144.3.788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Haigler H. T., McKanna J. A., Cohen S. Direct visualization of the binding and internalization of a ferritin conjugate of epidermal growth factor in human carcinoma cells A-431. J Cell Biol. 1979 May;81(2):382–395. doi: 10.1083/jcb.81.2.382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Haigler H. T., McKanna J. A., Cohen S. Rapid stimulation of pinocytosis in human carcinoma cells A-431 by epidermal growth factor. J Cell Biol. 1979 Oct;83(1):82–90. doi: 10.1083/jcb.83.1.82. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoffstein S., Soberman R., Goldstein I., Weissmann G. Concanavalin A induces microtubule assembly and specific granule discharge in human polymorphonuclear leukocytes. J Cell Biol. 1976 Mar;68(3):781–787. doi: 10.1083/jcb.68.3.781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jacobson K., Wenner C. E., Kemp G., Papahadjopoulos D. Surface properties of phorbol esters and their interaction with lipid monolayers and bilayers. Cancer Res. 1975 Nov;35(11 Pt 1):2991–2995. [PubMed] [Google Scholar]
  13. Johnston R. B., Jr, Godzik C. A., Cohn Z. A. Increased superoxide anion production by immunologically activated and chemically elicited macrophages. J Exp Med. 1978 Jul 1;148(1):115–127. doi: 10.1084/jem.148.1.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Klaus G. G. Cytochalasin B. Dissociation of pinocytosis and phagocytosis by peritoneal macrophages. Exp Cell Res. 1973 Apr;79(1):73–78. doi: 10.1016/0014-4827(73)90490-4. [DOI] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Lee L. S., Weinstein I. B. Tumor-promoting phorbol esters inhibit binding of epidermal growth factor to cellular receptors. Science. 1978 Oct 20;202(4365):313–315. doi: 10.1126/science.308698. [DOI] [PubMed] [Google Scholar]
  17. Lin D. C., Tobin K. D., Grumet M., Lin S. Cytochalasins inhibit nuclei-induced actin polymerization by blocking filament elongation. J Cell Biol. 1980 Feb;84(2):455–460. doi: 10.1083/jcb.84.2.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Loike J. D., Horwitz S. B. Effects of podophyllotoxin and VP-16-213 on microtubule assembly in vitro and nucleoside transport in HeLa cells. Biochemistry. 1976 Dec 14;15(25):5435–5443. doi: 10.1021/bi00670a003. [DOI] [PubMed] [Google Scholar]
  19. Mahoney E. M., Hamill A. L., Scott W. A., Cohn Z. A. Response of endocytosis to altered fatty acyl composition of macrophage phospholipids. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4895–4899. doi: 10.1073/pnas.74.11.4895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Michl J., Ohlbaum D. J., Silverstein S. C. 2-Deoxyglucose selectively inhibits Fc and complement receptor-mediated phagocytosis in mouse peritoneal macrophages. I. Description of the inhibitory effect. J Exp Med. 1976 Dec 1;144(6):1465–1483. doi: 10.1084/jem.144.6.1465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Mizel S. B., Wilson L. Nucleoside transport in mammalian cells. Inhibition by colchicine. Biochemistry. 1972 Jul 4;11(14):2573–2578. doi: 10.1021/bi00764a003. [DOI] [PubMed] [Google Scholar]
  23. Nathan C. F., Brukner L. H., Silverstein S. C., Cohn Z. A. Extracellular cytolysis by activated macrophages and granulocytes. I. Pharmacologic triggering of effector cells and the release of hydrogen peroxide. J Exp Med. 1979 Jan 1;149(1):84–99. doi: 10.1084/jem.149.1.84. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Orenstein J. M., Shelton E. Membrane phenomena accompanying erythrophagocytosis. A scanning electron microscope study. Lab Invest. 1977 Apr;36(4):363–374. [PubMed] [Google Scholar]
  25. Phaire-Washington L., Silverstein S. C., Wang E. Phorbol myristate acetate stimulates microtubule and 10-nm filament extension and lysosome redistribution in mouse macrophages. J Cell Biol. 1980 Aug;86(2):641–655. doi: 10.1083/jcb.86.2.641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Polliack A., Gordon S. Scanning electron microscopy of murine macrophages. Surface characteristics during maturation, activation, and phagocytosis. Lab Invest. 1975 Nov;33(5):469–477. [PubMed] [Google Scholar]
  27. Schlessinger J., Shechter Y., Willingham M. C., Pastan I. Direct visualization of binding, aggregation, and internalization of insulin and epidermal growth factor on living fibroblastic cells. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2659–2663. doi: 10.1073/pnas.75.6.2659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Silverstein S. C., Christman J. K., Acs G. The reovirus replicative cycle. Annu Rev Biochem. 1976;45:375–408. doi: 10.1146/annurev.bi.45.070176.002111. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Steinman R. M., Brodie S. E., Cohn Z. A. Membrane flow during pinocytosis. A stereologic analysis. J Cell Biol. 1976 Mar;68(3):665–687. doi: 10.1083/jcb.68.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Steinman R. M., Cohn Z. A. The interaction of particulate horseradish peroxidase (HRP)-anti HRP immune complexes with mouse peritoneal macrophages in vitro. J Cell Biol. 1972 Dec;55(3):616–634. doi: 10.1083/jcb.55.3.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Steinman R. M., Cohn Z. A. The interaction of soluble horseradish peroxidase with mouse peritoneal macrophages in vitro. J Cell Biol. 1972 Oct;55(1):186–204. doi: 10.1083/jcb.55.1.186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Steinman R. M., Silver J. M., Cohn Z. A. Pinocytosis in fibroblasts. Quantitative studies in vitro. J Cell Biol. 1974 Dec;63(3):949–969. doi: 10.1083/jcb.63.3.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Vassalli J. D., Hamilton J., Reich E. Macrophage plasminogen activator: induction by concanavalin A and phorbol myristate acetate. Cell. 1977 Jul;11(3):695–705. doi: 10.1016/0092-8674(77)90086-1. [DOI] [PubMed] [Google Scholar]
  35. Wagner R., Rosenberg M., Estensen R. Endocytosis in Chang liver cells. Quantitation by sucrose- 3 H uptake and inhibition by cytochalasin B. J Cell Biol. 1971 Sep;50(3):804–817. doi: 10.1083/jcb.50.3.804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weihing R. R. Cytochalasin B inhibits actin-related gelation of HeLa cell extracts. J Cell Biol. 1976 Oct;71(1):303–307. doi: 10.1083/jcb.71.1.303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Wigler M., Weinstein I. B. Tumour promotor induces plasminogen activator. Nature. 1976 Jan 22;259(5540):232–233. doi: 10.1038/259232a0. [DOI] [PubMed] [Google Scholar]
  38. Wills E. J., Davies P., Allison A. C., Haswell A. D. Cytochalasin B fails to inhibit pinocytosis by macrophages. Nat New Biol. 1972 Nov 8;240(97):58–60. doi: 10.1038/newbio240058a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES