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. 1991 Feb 1;112(3):407–425. doi: 10.1083/jcb.112.3.407

The cytoskeleton of the resting human blood platelet: structure of the membrane skeleton and its attachment to actin filaments

PMCID: PMC2288836  PMID: 1991790

Abstract

We used high-resolution EM and immunocytochemistry in combination with different specimen preparation techniques to resolve the ultrastructure of the resting platelet cytoskeleton. The periphery of the cytoskeleton, an electron-dense subplasmalemmal region in thin section electron micrographs, is a tightly woven planar sheet composed of a spectrin-rich network whose interstices contain GPIb/IX-actin-binding protein (ABP) complexes. This membrane skeleton connects to a system of curved actin filaments (F-actin) that emanate from a central oval core of F-actin cross-linked by ABP. The predominant interaction of the radial actin filaments with the membrane skeleton is along their sides, and the strongest connection between the membrane skeleton and F-actin is via ABP-GPIb ligands, although there is evidence for spectrin attaching to the ends of the radial actin filaments as well. Since a mechanical separation of the F-actin cores and radial F-actin-GPIb-ABP complexes from the underlying spectrin-rich skeleton leads to the latter's expansion, it follows that the spectrin-based skeleton of the resting cell may be held in a compressed form by interdigitating GPIb/IX complexes which are immobilized by radial F-actin-ABP anchors.

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Selected References

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  1. Boyles J., Fox J. E., Phillips D. R., Stenberg P. E. Organization of the cytoskeleton in resting, discoid platelets: preservation of actin filaments by a modified fixation that prevents osmium damage. J Cell Biol. 1985 Oct;101(4):1463–1472. doi: 10.1083/jcb.101.4.1463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ezzell R. M., Kenney D. M., Egan S., Stossel T. P., Hartwig J. H. Localization of the domain of actin-binding protein that binds to membrane glycoprotein Ib and actin in human platelets. J Biol Chem. 1988 Sep 15;263(26):13303–13309. [PubMed] [Google Scholar]
  3. Fox J. E., Aggerbeck L. P., Berndt M. C. Structure of the glycoprotein Ib.IX complex from platelet membranes. J Biol Chem. 1988 Apr 5;263(10):4882–4890. [PubMed] [Google Scholar]
  4. Fox J. E., Boyles J. K., Berndt M. C., Steffen P. K., Anderson L. K. Identification of a membrane skeleton in platelets. J Cell Biol. 1988 May;106(5):1525–1538. doi: 10.1083/jcb.106.5.1525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fox J. E. Identification of actin-binding protein as the protein linking the membrane skeleton to glycoproteins on platelet plasma membranes. J Biol Chem. 1985 Oct 5;260(22):11970–11977. [PubMed] [Google Scholar]
  6. Fox J. E., Reynolds C. C., Morrow J. S., Phillips D. R. Spectrin is associated with membrane-bound actin filaments in platelets and is hydrolyzed by the Ca2+-dependent protease during platelet activation. Blood. 1987 Feb;69(2):537–545. [PubMed] [Google Scholar]
  7. George J. N., Pickett E. B., Saucerman S., McEver R. P., Kunicki T. J., Kieffer N., Newman P. J. Platelet surface glycoproteins. Studies on resting and activated platelets and platelet membrane microparticles in normal subjects, and observations in patients during adult respiratory distress syndrome and cardiac surgery. J Clin Invest. 1986 Aug;78(2):340–348. doi: 10.1172/JCI112582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Glenney J. R., Jr, Glenney P., Weber K. Erythroid spectrin, brain fodrin, and intestinal brush border proteins (TW-260/240) are related molecules containing a common calmodulin-binding subunit bound to a variant cell type-specific subunit. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4002–4005. doi: 10.1073/pnas.79.13.4002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hartwig J. H., Chambers K. A., Hopcia K. L., Kwiatkowski D. J. Association of profilin with filament-free regions of human leukocyte and platelet membranes and reversible membrane binding during platelet activation. J Cell Biol. 1989 Oct;109(4 Pt 1):1571–1579. doi: 10.1083/jcb.109.4.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hartwig J. H., Chambers K. A., Stossel T. P. Association of gelsolin with actin filaments and cell membranes of macrophages and platelets. J Cell Biol. 1989 Feb;108(2):467–479. doi: 10.1083/jcb.108.2.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hartwig J. H., Shevlin P. The architecture of actin filaments and the ultrastructural location of actin-binding protein in the periphery of lung macrophages. J Cell Biol. 1986 Sep;103(3):1007–1020. doi: 10.1083/jcb.103.3.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lassing I., Lindberg U. Evidence that the phosphatidylinositol cycle is linked to cell motility. Exp Cell Res. 1988 Jan;174(1):1–15. doi: 10.1016/0014-4827(88)90136-x. [DOI] [PubMed] [Google Scholar]
  14. Lind S. E., Janmey P. A., Chaponnier C., Herbert T. J., Stossel T. P. Reversible binding of actin to gelsolin and profilin in human platelet extracts. J Cell Biol. 1987 Aug;105(2):833–842. doi: 10.1083/jcb.105.2.833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Liu S. C., Derick L. H., Palek J. Visualization of the hexagonal lattice in the erythrocyte membrane skeleton. J Cell Biol. 1987 Mar;104(3):527–536. doi: 10.1083/jcb.104.3.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Loftus J. C., Choate J., Albrecht R. M. Platelet activation and cytoskeletal reorganization: high voltage electron microscopic examination of intact and Triton-extracted whole mounts. J Cell Biol. 1984 Jun;98(6):2019–2025. doi: 10.1083/jcb.98.6.2019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McGough A. M., Josephs R. On the structure of erythrocyte spectrin in partially expanded membrane skeletons. Proc Natl Acad Sci U S A. 1990 Jul;87(13):5208–5212. doi: 10.1073/pnas.87.13.5208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Michelson A. D., Barnard M. R. Thrombin-induced changes in platelet membrane glycoproteins Ib, IX, and IIb-IIIa complex. Blood. 1987 Nov;70(5):1673–1678. [PubMed] [Google Scholar]
  19. Morrow J. S. The spectrin membrane skeleton: emerging concepts. Curr Opin Cell Biol. 1989 Feb;1(1):23–29. doi: 10.1016/s0955-0674(89)80032-8. [DOI] [PubMed] [Google Scholar]
  20. Nachmias V. T. Cytoskeleton of human platelets at rest and after spreading. J Cell Biol. 1980 Sep;86(3):795–802. doi: 10.1083/jcb.86.3.795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nachmias V. T., Sullender J., Fallon J., Asch A. Observations on the "cytoskeleton" of human platelets. Thromb Haemost. 1980 Feb 29;42(5):1661–1666. [PubMed] [Google Scholar]
  22. Nakata T., Hirokawa N. Cytoskeletal reorganization of human platelets after stimulation revealed by the quick-freeze deep-etch technique. J Cell Biol. 1987 Oct;105(4):1771–1780. doi: 10.1083/jcb.105.4.1771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. O'Halloran T., Beckerle M. C., Burridge K. Identification of talin as a major cytoplasmic protein implicated in platelet activation. Nature. 1985 Oct 3;317(6036):449–451. doi: 10.1038/317449a0. [DOI] [PubMed] [Google Scholar]
  24. Okita J. R., Pidard D., Newman P. J., Montgomery R. R., Kunicki T. J. On the association of glycoprotein Ib and actin-binding protein in human platelets. J Cell Biol. 1985 Jan;100(1):317–321. doi: 10.1083/jcb.100.1.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pinder J. C., Weeds A. G., Gratzer W. B. Study of actin filament ends in the human red cell membrane. J Mol Biol. 1986 Oct 5;191(3):461–468. doi: 10.1016/0022-2836(86)90141-5. [DOI] [PubMed] [Google Scholar]
  26. Rosenberg S., Stracher A., Lucas R. C. Isolation and characterization of actin and actin-binding protein from human platelets. J Cell Biol. 1981 Oct;91(1):201–211. doi: 10.1083/jcb.91.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Safer D., Golla R., Nachmias V. T. Isolation of a 5-kilodalton actin-sequestering peptide from human blood platelets. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2536–2540. doi: 10.1073/pnas.87.7.2536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schollmeyer J. V., Rao G. H., White J. G. An actin-binding protein in human platelets. Interactions with alpha-actinin on gelatin of actin and the influence of cytochalasin B. Am J Pathol. 1978 Nov;93(2):433–446. [PMC free article] [PubMed] [Google Scholar]
  29. Shen B. W., Josephs R., Steck T. L. Ultrastructure of the intact skeleton of the human erythrocyte membrane. J Cell Biol. 1986 Mar;102(3):997–1006. doi: 10.1083/jcb.102.3.997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sixma J. J., van den Berg A., Jockusch B. M., Hartwig J. Immunoelectron microscopic localization of actin, alpha-actinin, actin-binding protein and myosin in resting and activated human blood platelets. Eur J Cell Biol. 1989 Apr;48(2):271–281. [PubMed] [Google Scholar]
  31. Slot J. W., Geuze H. J. A new method of preparing gold probes for multiple-labeling cytochemistry. Eur J Cell Biol. 1985 Jul;38(1):87–93. [PubMed] [Google Scholar]
  32. Stendahl O. I., Hartwig J. H., Brotschi E. A., Stossel T. P. Distribution of actin-binding protein and myosin in macrophages during spreading and phagocytosis. J Cell Biol. 1980 Feb;84(2):215–224. doi: 10.1083/jcb.84.2.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Takeuchi K., Kuroda K., Ishigami M., Nakamura T. Actin cytoskeleton of resting bovine platelets. Exp Cell Res. 1990 Feb;186(2):374–380. doi: 10.1016/0014-4827(90)90319-6. [DOI] [PubMed] [Google Scholar]
  35. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tyler J. M., Anderson J. M., Branton D. Structural comparison of several actin-binding macromolecules. J Cell Biol. 1980 May;85(2):489–495. doi: 10.1083/jcb.85.2.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. White J. G. Arrangements of actin filaments in the cytoskeleton of human platelets. Am J Pathol. 1984 Nov;117(2):207–217. [PMC free article] [PubMed] [Google Scholar]
  38. Yin H. L., Kwiatkowski D. J., Mole J. E., Cole F. S. Structure and biosynthesis of cytoplasmic and secreted variants of gelsolin. J Biol Chem. 1984 Apr 25;259(8):5271–5276. [PubMed] [Google Scholar]

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