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. 1979 Aug 1;82(2):347–368. doi: 10.1083/jcb.82.2.347

Changing patterns of plasma membrane-associated filaments during the initial phases of polymorphonuclear leukocyte adherence

PMCID: PMC2110465  PMID: 383726

Abstract

By utilizing a combination of several ultrastructural techniques, we have been able to demonstrate differences in filament organization on the adherent plasma membranes of spreading and mobile PMN as well as within the extending lamellipodia. To follow the subplasmalemmal filaments of this small amoeboid cell during these kinetic events, we sheared off the upper portions of cells onto glass and carbon surfaces for 30 s--5 min. The exposed adherent membranes were immediately fixed and processed for high-resolution SEM or TEM. Whole cells were also examined by phase contrast microscopy, SEM, and oriented thin sections. Observed by SEM, the inner surface of nonadherent PMN membranes is free of filaments, but within 30 s of attachment to the substrate a three- dimensional, interlocking network of globular projections and radiating microfilaments--i.e., a subplasmalemmal filament complex--is consistently demonstrable (with or without postfixation in OsO4). Seen by TEM, extending lamellipodia contain a felt of filamentous and finely granular material, distinct from the golbule/filament complex of the adjacent adherent membrane. In the spread cell, this golbule-filament complex covers the entire lower membrane and increases in filament- density over the next 2--3 min. By 3--5 min after plating, as the PMN rounds up before the initiation of amoeboid movements, another pattern emerges--circumferential bands of anastomosing filament bundles in which thick, short filaments resembling myosin are found. This work provides structural evidence on the organization of polymerized contractile elements associated with the plasma membrane during cellular adherence.

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Selected References

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

  1. Abercrombie M., Heaysman J. E., Pegrum S. M. The locomotion of fibroblasts in culture. IV. Electron microscopy of the leading lamella. Exp Cell Res. 1971 Aug;67(2):359–367. doi: 10.1016/0014-4827(71)90420-4. [DOI] [PubMed] [Google Scholar]
  2. Boxer L. A., Hedley-Whyte E. T., Stossel T. P. Neutrophil action dysfunction and abnormal neutrophil behavior. N Engl J Med. 1974 Nov 21;291(21):1093–1099. doi: 10.1056/NEJM197411212912101. [DOI] [PubMed] [Google Scholar]
  3. Boxer L. A., Richardson S., Floyd A. Identification of actin-binding protein in membrane of polymorphonuclear leukocytes. Nature. 1976 Sep 16;263(5574):249–251. doi: 10.1038/263249a0. [DOI] [PubMed] [Google Scholar]
  4. Boxer L. A., Stossel T. P. Interactions of actin, myosin, and an actin-binding protein of chronic myelogenous leukemia leukocytes. J Clin Invest. 1976 Apr;57(4):964–976. doi: 10.1172/JCI108373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buckley I. K., Raju T. R. Form and distribution of actin and myosin in non-muscle cells: a study using cultured chick embryo fibroblasts. J Microsc. 1976 Jul;107(2):129–149. doi: 10.1111/j.1365-2818.1976.tb02431.x. [DOI] [PubMed] [Google Scholar]
  6. Buckley I. K. Three dimensional fine structure of cultured cells: possible implications for subcellular motility. Tissue Cell. 1975;7(1):51–72. doi: 10.1016/s0040-8166(75)80007-3. [DOI] [PubMed] [Google Scholar]
  7. Chang C. M., Goldman R. D. The localization of actin-like fibers in cultured neuroblastoma cells as revealed by heavy meromyosin binding. J Cell Biol. 1973 Jun;57(3):867–874. doi: 10.1083/jcb.57.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clarke M., Schatten G., Mazia D., Spudich J. A. Visualization of actin fibers associated with the cell membrane in amoebae of Dictyostelium discoideum. Proc Natl Acad Sci U S A. 1975 May;72(5):1758–1762. doi: 10.1073/pnas.72.5.1758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Condeelis J. S., Taylor D. L. The contractile basis of amoeboid movement. V. The control of gelation, solation, and contraction in extracts from Dictyostelium discoideum. J Cell Biol. 1977 Sep;74(3):901–927. doi: 10.1083/jcb.74.3.901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Goldman R. D. The use of heavy meromyosin binding as an ultrastructural cytochemical method for localizing and determining the possible functions of actin-like microfilaments in nonmuscle cells. J Histochem Cytochem. 1975 Jul;23(7):529–542. doi: 10.1177/23.7.1095652. [DOI] [PubMed] [Google Scholar]
  11. Heaysman J. E., Pegrum S. M. Early contacts between fibroblasts. An ultrastructural study. Exp Cell Res. 1973 Mar 30;78(1):71–78. doi: 10.1016/0014-4827(73)90039-6. [DOI] [PubMed] [Google Scholar]
  12. Heggeness M. H., Wang K., Singer S. J. Intracellular distributions of mechanochemical proteins in cultured fibroblasts. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3883–3887. doi: 10.1073/pnas.74.9.3883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hitchcock S. E. Regulation of motility in nonmuscle cells. J Cell Biol. 1977 Jul;74(1):1–15. doi: 10.1083/jcb.74.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Korn E. D. Biochemistry of actomyosin-dependent cell motility (a review). Proc Natl Acad Sci U S A. 1978 Feb;75(2):588–599. doi: 10.1073/pnas.75.2.588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lazarides E., Burridge K. Alpha-actinin: immunofluorescent localization of a muscle structural protein in nonmuscle cells. Cell. 1975 Nov;6(3):289–298. doi: 10.1016/0092-8674(75)90180-4. [DOI] [PubMed] [Google Scholar]
  16. Lazarides E. Immunofluorescence studies on the structure of actin filaments in tissue culture cells. J Histochem Cytochem. 1975 Jul;23(7):507–528. doi: 10.1177/23.7.1095651. [DOI] [PubMed] [Google Scholar]
  17. Maupin-Szamier P., Pollard T. D. Actin filament destruction by osmium tetroxide. J Cell Biol. 1978 Jun;77(3):837–852. doi: 10.1083/jcb.77.3.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McNutt N. S., Culp L. A., Black P. H. Contact-inhibited revertant cell lines isolated from SV40-transformed cells. II. Ultrastructural study. J Cell Biol. 1971 Sep;50(3):691–708. doi: 10.1083/jcb.50.3.691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pegrum S. M., Maroudas N. G. Early events in fibroblast adhesion to glass. An electron microscopic study. Exp Cell Res. 1975 Dec;96(2):416–422. doi: 10.1016/0014-4827(75)90277-3. [DOI] [PubMed] [Google Scholar]
  20. Pollard T. D. Cytoskeletal functions of cytoplasmic contractile proteins. J Supramol Struct. 1976;5(3):317–334. doi: 10.1002/jss.400050306. [DOI] [PubMed] [Google Scholar]
  21. Pollard T. D. The role of actin in the temperature-dependent gelation and contraction of extracts of Acanthamoeba. J Cell Biol. 1976 Mar;68(3):579–601. doi: 10.1083/jcb.68.3.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pollard T. D., Weihing R. R. Actin and myosin and cell movement. CRC Crit Rev Biochem. 1974 Jan;2(1):1–65. doi: 10.3109/10409237409105443. [DOI] [PubMed] [Google Scholar]
  23. Rikihisa Y., Mizuno D. Demonstration of myosin on the cytoplasmic side of plasma membranes of guinea pig polymorphonuclear leukocytes with immunoferritin. Exp Cell Res. 1977 Nov;110(1):87–92. doi: 10.1016/0014-4827(77)90273-7. [DOI] [PubMed] [Google Scholar]
  24. Sanger J. W. Intracellular localization of actin with fluorescently labelled heavy meromyosin. Cell Tissue Res. 1975 Aug 27;161(4):431–434. doi: 10.1007/BF00224134. [DOI] [PubMed] [Google Scholar]
  25. Sawada H., Ishikawa H., Yamada E. High resolution scanning electron microscopy of frog sartorius muscle. Tissue Cell. 1978;10(1):179–190. doi: 10.1016/0040-8166(78)90016-2. [DOI] [PubMed] [Google Scholar]
  26. Schreiner G. F., Fujiwara K., Pollard T. D., Unanue E. R. Redistribution of myosin accompanying capping of surface Ig. J Exp Med. 1977 May 1;145(5):1393–1398. doi: 10.1084/jem.145.5.1393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Senda N., Tamura H., Shibata N., Yoshitake J., Konko K., Tanaka K. The mechanism of the movement of leucocytes. Exp Cell Res. 1975 Mar 15;91(2):393–407. doi: 10.1016/0014-4827(75)90120-2. [DOI] [PubMed] [Google Scholar]
  28. Shibata N., Tatsumi N., Tanaka K., Okamura Y., Senda N. Leucocyte myosin and its location in the cell. Biochim Biophys Acta. 1975 Aug 19;400(2):222–243. doi: 10.1016/0005-2795(75)90177-4. [DOI] [PubMed] [Google Scholar]
  29. Stossel T. P., Hartwig J. H. Interactions of actin, myosin, and a new actin-binding protein of rabbit pulmonary macrophages. II. Role in cytoplasmic movement and phagocytosis. J Cell Biol. 1976 Mar;68(3):602–619. doi: 10.1083/jcb.68.3.602. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Stossel T. P., Pollard T. D. Myosin in polymorphonuclear leukocytes. J Biol Chem. 1973 Dec 10;248(23):8288–8294. [PubMed] [Google Scholar]
  31. Taylor D. L., Rhodes J. A., Hammond S. A. The contractile basis of ameboid movement. II. Structure and contractility of motile extracts and plasmalemma-ectoplasm ghosts. J Cell Biol. 1976 Jul;70(1):123–143. doi: 10.1083/jcb.70.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vacquier V. D. The isolation of intact cortical granules from sea urchin eggs: calcium lons trigger granule discharge. Dev Biol. 1975 Mar;43(1):62–74. doi: 10.1016/0012-1606(75)90131-1. [DOI] [PubMed] [Google Scholar]
  33. Wang K., Singer S. J. Interaction of filamin with f-actin in solution. Proc Natl Acad Sci U S A. 1977 May;74(5):2021–2025. doi: 10.1073/pnas.74.5.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wolosewick J. J., Porter K. R. Stereo high-voltage electron microscopy of whole cells of the human diploid line, WI-38. Am J Anat. 1976 Nov;147(3):303–323. doi: 10.1002/aja.1001470305. [DOI] [PubMed] [Google Scholar]
  35. Yamada K. M., Spooner B. S., Wessells N. K. Ultrastructure and function of growth cones and axons of cultured nerve cells. J Cell Biol. 1971 Jun;49(3):614–635. doi: 10.1083/jcb.49.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zucker-Franklin D. Electron microscopic studies of human granulocytes: structural variations related to function. Semin Hematol. 1968 Apr;5(2):109–133. [PubMed] [Google Scholar]

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