Skip to main content
NCBI home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1990 Jan;136(1):61–70.

Plasma membrane GPIIb/IIIa. Evidence for a cycling receptor pool.

J D Wencel-Drake 1
PMCID: PMC1877449  PMID: 2404419

Abstract

The author used immunofluorescence and digital image processing to investigate the dynamic distribution of GPIIb/IIIa in living platelets. Resting cells were incubated with AP-2, a complex-specific, monoclonal, anti-GPIIb/IIIa antibody. Examination of intact cells demonstrated a rim pattern for GPIIb/IIIa consistent with a surface localization. Permeabilization revealed a time-dependent increase in the labeling of apparent intracellular vacuoles. This pattern is distinct from the "patch-cap" pattern observed when unfixed platelets were incubated with fluoresceinated concanavalin A. Additionally, labeling of this vacuolar pool of GPIIb/IIIa was inhibited by treatment with 2% sodium azide or by incubation at 4 degrees C. Identical staining patterns were obtained with Fab fragments of AP-2. Ultrastructural examination confirmed the presence of labeled intracellular vacuolar structures. Parallel studies performed with AP-1, a monoclonal anti-GPIb antibody, failed to demonstrate internalization of GPIb. Finally, thrombin stimulation of resting platelets, which had been preincubated with AP-2, resulted in the clearing of this newly internalized pool of GPIIb/IIIa; presumably via translocation to the surface. These data suggest the presence of an actively cycling pool of GPIIb/IIIa that has not been described previously. The dynamic distribution of this pool may be important in the regulation of platelet adhesiveness.

Full text

PDF
61

Images in this article

62Figure 1
on p.62
63Figure 2
on p.63
64Figure 3
on p.64
65Figure 4
on p.65
65Figure 5
on p.65
66Figure 6
on p.66
67Figure 7
on p.67
68Figure 8
on p.68

Selected References

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

  1. Ackerman G. A., Wolken K. W. Histochemical evidence for the differential surface labeling, uptake, and intracellular transport of a colloidal gold-labeled insulin complex by normal human blood cells. J Histochem Cytochem. 1981 Oct;29(10):1137–1149. doi: 10.1177/29.10.6271866. [DOI] [PubMed] [Google Scholar]
  2. Albrecht R. M., Goodman S. L., Simmons S. R. Distribution and movement of membrane-associated platelet glycoproteins: use of colloidal gold with correlative video-enhanced light microscopy, low-voltage high-resolution scanning electron microscopy, and high-voltage transmission electron microscopy. Am J Anat. 1989 Jun-Jul;185(2-3):149–164. doi: 10.1002/aja.1001850208. [DOI] [PubMed] [Google Scholar]
  3. Behnke O., Bray D. Surface movements during the spreading of blood platelets. Eur J Cell Biol. 1988 Jun;46(2):207–216. [PubMed] [Google Scholar]
  4. Behnke O. Surface membrane clearing of receptor-ligand complexes in human blood platelets. J Cell Sci. 1987 Apr;87(Pt 3):465–472. doi: 10.1242/jcs.87.3.465. [DOI] [PubMed] [Google Scholar]
  5. Bourguignon L. Y. Receptor capping in platelet membranes. Cell Biol Int Rep. 1984 Jan;8(1):19–26. doi: 10.1016/0309-1651(84)90177-2. [DOI] [PubMed] [Google Scholar]
  6. Fitzgerald L. A., Phillips D. R. Calcium regulation of the platelet membrane glycoprotein IIb-IIIa complex. J Biol Chem. 1985 Sep 15;260(20):11366–11374. [PubMed] [Google Scholar]
  7. 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]
  8. Ginsberg M. H., Painter R. G., Forsyth J., Birdwell C., Plow E. F. Thrombin increases expression of fibronectin antigen on the platelet surface. Proc Natl Acad Sci U S A. 1980 Feb;77(2):1049–1053. doi: 10.1073/pnas.77.2.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Ginsberg M. H., Taylor L., Painter R. G. The mechanism of thrombin-induced platelet factor 4 secretion. Blood. 1980 Apr;55(4):661–668. [PubMed] [Google Scholar]
  10. Hoshikawa N., Ashihara Y., Shirasawa K. Studies on the initial reactive sites for platelet activation. Cationized ferritin- and wheat germ agglutinin-binding sites on the cells. Acta Pathol Jpn. 1985 Sep;35(5):1069–1085. doi: 10.1111/j.1440-1827.1985.tb00999.x. [DOI] [PubMed] [Google Scholar]
  11. Kakaiya R. M., Kiraly T. L., Cable R. G. Concanavalin A induces patching/capping of the platelet membrane glycoprotein IIb/IIIa complex. Thromb Haemost. 1988 Apr 8;59(2):281–283. [PubMed] [Google Scholar]
  12. Kawakami H., Hirano H. Rearrangement of the open-canalicular system of the human blood platelet after incorporation of surface-bound ligands. A high-voltage electron-microscopic study. Cell Tissue Res. 1986;245(3):465–469. doi: 10.1007/BF00218545. [DOI] [PubMed] [Google Scholar]
  13. Kemona H., Andrzejewska A., Prokopowicz J., Nowak H., Mantur M. Phagocytic activity of human blood platelets examined by electron microscopy. Folia Haematol Int Mag Klin Morphol Blutforsch. 1986;113(5):696–702. [PubMed] [Google Scholar]
  14. Kunicki T. J., Nugent D. J., Piotrowicz R. S., Lai C. S. Covalent attachment of sulfhydryl-specific, electron spin resonance spin-labels to Fab' fragments of murine monoclonal antibodies that recognize human platelet membrane glycoproteins. Development of membrane protein specific spin probes. Biochemistry. 1986 Sep 9;25(18):4979–4983. doi: 10.1021/bi00366a001. [DOI] [PubMed] [Google Scholar]
  15. Lewis J. C., Maldonado J. E., Mann K. G. Phagocytosis in human platelets: localization of acid phosphatase-positive phagosomes following latex uptake. Blood. 1976 May;47(5):833–840. [PubMed] [Google Scholar]
  16. Loftus J. C., Albrecht R. M. Redistribution of the fibrinogen receptor of human platelets after surface activation. J Cell Biol. 1984 Sep;99(3):822–829. doi: 10.1083/jcb.99.3.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Martin-Nizard F., Richard B., Torpier G., Nouvelot A., Fruchart J. C., Duthilleul P., Delbart C. Analysis of phospholipid transfer during HDL binding to platelets using a fluorescent analog of phosphatidylcholine. Thromb Res. 1987 Jun 15;46(6):811–825. doi: 10.1016/0049-3848(87)90073-9. [DOI] [PubMed] [Google Scholar]
  18. Montgomery R. R., Kunicki T. J., Taves C., Pidard D., Corcoran M. Diagnosis of Bernard-Soulier syndrome and Glanzmann's thrombasthenia with a monoclonal assay on whole blood. J Clin Invest. 1983 Feb;71(2):385–389. doi: 10.1172/JCI110780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Morgenstern E., Neumann K., Patscheke H. The exocytosis of human blood platelets. A fast freezing and freeze-substitution analysis. Eur J Cell Biol. 1987 Apr;43(2):273–282. [PubMed] [Google Scholar]
  20. 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]
  21. Pidard D., Montgomery R. R., Bennett J. S., Kunicki T. J. Interaction of AP-2, a monoclonal antibody specific for the human platelet glycoprotein IIb-IIIa complex, with intact platelets. J Biol Chem. 1983 Oct 25;258(20):12582–12586. [PubMed] [Google Scholar]
  22. Plow E. F., Loftus J. C., Levin E. G., Fair D. S., Dixon D., Forsyth J., Ginsberg M. H. Immunologic relationship between platelet membrane glycoprotein GPIIb/IIIa and cell surface molecules expressed by a variety of cells. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6002–6006. doi: 10.1073/pnas.83.16.6002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Salisbury J. L., Condeelis J. S., Satir P. Receptor-mediated endocytosis: machinery and regulation of the clathrin-coated vesicle pathway. Int Rev Exp Pathol. 1983;24:1–62. [PubMed] [Google Scholar]
  24. Santoso S., Zimmermann U., Neppert J., Mueller-Eckhardt C. Receptor patching and capping of platelet membranes induced by monoclonal antibodies. Blood. 1986 Feb;67(2):343–349. [PubMed] [Google Scholar]
  25. Shattil S. J., Hoxie J. A., Cunningham M., Brass L. F. Changes in the platelet membrane glycoprotein IIb.IIIa complex during platelet activation. J Biol Chem. 1985 Sep 15;260(20):11107–11114. [PubMed] [Google Scholar]
  26. Sixma J. J., van den Berg A., Schiphorst M., Geuze H. J., McDonagh J. Immunocytochemical localization of albumin and factor XIII in thin cryo sections of human blood platelets. Thromb Haemost. 1984 Jul 29;51(3):388–391. [PubMed] [Google Scholar]
  27. Spycher M. O., Nydegger U. E. Part of the activating cross-linked immunoglobulin G is internalized by human platelets to sites not accessible for enzymatic digestion. Blood. 1986 Jan;67(1):12–18. [PubMed] [Google Scholar]
  28. Suzuki H., Tanoue K., Yamazaki H. Endocytosis by platelets during cationized ferritin-induced aggregation. Cell Tissue Res. 1985;240(3):513–517. doi: 10.1007/BF00216339. [DOI] [PubMed] [Google Scholar]
  29. Wencel-Drake J. D., Painter R. G., Zimmerman T. S., Ginsberg M. H. Ultrastructural localization of human platelet thrombospondin, fibrinogen, fibronectin, and von Willebrand factor in frozen thin section. Blood. 1985 Apr;65(4):929–938. [PubMed] [Google Scholar]
  30. Wencel-Drake J. D., Plow E. F., Kunicki T. J., Woods V. L., Keller D. M., Ginsberg M. H. Localization of internal pools of membrane glycoproteins involved in platelet adhesive responses. Am J Pathol. 1986 Aug;124(2):324–334. [PMC free article] [PubMed] [Google Scholar]
  31. Wencel-Drake J. D., Plow E. F., Zimmerman T. S., Painter R. G., Ginsberg M. H. Immunofluorescent localization of adhesive glycoproteins in resting and thrombin-stimulated platelets. Am J Pathol. 1984 May;115(2):156–164. [PMC free article] [PubMed] [Google Scholar]
  32. White J. G. A search for the platelet secretory pathway using electron dense tracers. Am J Pathol. 1970 Jan;58(1):31–49. [PMC free article] [PubMed] [Google Scholar]
  33. White J. G., Clawson C. C. Effects of large latex particle uptake of the surface connected canalicular system of blood platelets: a freeze-fracture and cytochemical study. Ultrastruct Pathol. 1981 Jul-Sep;2(3):277–287. doi: 10.3109/01913128109048311. [DOI] [PubMed] [Google Scholar]
  34. White J. G., Clawson C. C. Effects of small latex particle uptake on the surface connected canalicular system of blood platelets: a freeze-fracture and cytochemical study. Diagn Histopathol. 1982 Jan-Mar;5(1):3–10. [PubMed] [Google Scholar]
  35. White J. G. Electron microscopic studies of platelet secretion. Prog Hemost Thromb. 1974;2(0):49–98. [PubMed] [Google Scholar]
  36. White J. G., Krumwiede M. Further studies of the secretory pathway in thrombin-stimulated human platelets. Blood. 1987 Apr;69(4):1196–1203. [PubMed] [Google Scholar]
  37. White J. G. The transfer of thorium particles from plasma to platelets and platelet granules. Am J Pathol. 1968 Oct;53(4):567–575. [PMC free article] [PubMed] [Google Scholar]
  38. Yamazaki H., Suzuki H., Yamamoto N., Tanoue K. Electron microscopic observations on platelet aggregation induced by cationized ferritin. Blood. 1984 Feb;63(2):439–447. [PubMed] [Google Scholar]
  39. Zucker-Franklin D. Endocytosis by human platelets: metabolic and freeze-fracture studies. J Cell Biol. 1981 Dec;91(3 Pt 1):706–715. doi: 10.1083/jcb.91.3.706. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES