Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1989 Jul;84(1):73–82. doi: 10.1172/JCI114173

Incorporation of intravenously injected albumin, immunoglobulin G, and fibrinogen in guinea pig megakaryocyte granules.

P J Handagama 1, M A Shuman 1, D F Bainton 1
PMCID: PMC303955  PMID: 2738161

Abstract

In a previous study we provide evidence for a circuitous pathway by which circulating plasma proteins enter megakaryocyte granules by an endocytic mechanism and are returned to the circulation in platelets (1987. Proc. Natl. Acad. Sci. USA. 84:861-865). Horseradish peroxidase (40,000 mol wt) was injected into guinea pigs and its uptake into megakaryocyte organelles examined by electron microscopy and cytochemistry. In the present study we tested the ability of guinea pig megakaryocytes to take up intravenously injected albumin, IgG, and fibrinogen. We used two types of proteins to study the endocytic pathway: (a) heterologous human proteins, which were detected immunohistochemically using antibodies that do not crossreact with the native guinea pig counterparts; and (b) human and guinea pig proteins labeled with the small (250 mol wt), inert molecule, biotin, which were detected using an antibody against biotin. We detected all three of the injected proteins in bone marrow megakaryocytes in patterns identical to those of native counterparts. The injected protein consistently appeared in platelets 24 h later and was secreted in response to thrombin. We conclude that there are at least two mechanisms by which guinea pig megakaryocyte granules acquire proteins (a) endogenous synthesis, as demonstrated by others, and (b) endocytosis of plasma proteins synthesized by other types of cells.

Full text

PDF
73

Images in this article

Selected References

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

  1. Bainton D. F., Miller L. J., Kishimoto T. K., Springer T. A. Leukocyte adhesion receptors are stored in peroxidase-negative granules of human neutrophils. J Exp Med. 1987 Dec 1;166(6):1641–1653. doi: 10.1084/jem.166.6.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beckstead J. H., Stenberg P. E., McEver R. P., Shuman M. A., Bainton D. F. Immunohistochemical localization of membrane and alpha-granule proteins in human megakaryocytes: application to plastic-embedded bone marrow biopsy specimens. Blood. 1986 Feb;67(2):285–293. [PubMed] [Google Scholar]
  3. Belloc F., Hourdille P., Fialon P., Boisseau M. R., Soria J. Fibrinogen synthesis by megakaryocyte rich human marrow cell concentrates. Thromb Res. 1985 May 15;38(4):341–351. doi: 10.1016/0049-3848(85)90133-1. [DOI] [PubMed] [Google Scholar]
  4. Castaldi P. A., Caen J. Platelet fibrinogen. J Clin Pathol. 1965 Sep;18(5):579–585. doi: 10.1136/jcp.18.5.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chiu H. C., Schick P. K., Colman R. W. Biosynthesis of factor V in isolated guinea pig megakaryocytes. J Clin Invest. 1985 Feb;75(2):339–346. doi: 10.1172/JCI111706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cramer E. M., Meyer D., le Menn R., Breton-Gorius J. Eccentric localization of von Willebrand factor in an internal structure of platelet alpha-granule resembling that of Weibel-Palade bodies. Blood. 1985 Sep;66(3):710–713. [PubMed] [Google Scholar]
  7. De Bruyn P. P., Michelson S., Bankston P. W. In-vivo endocytosis by bristle-coated pits and intracellular transport of endogenous albumin in the endothelium of the sinuses of liver and bone marrow. Cell Tissue Res. 1985;240(1):1–7. doi: 10.1007/BF00217551. [DOI] [PubMed] [Google Scholar]
  8. Doolittle R. F., Takagi T., Cottrell B. A. Platelet and plasma fibrinogens are identical gene products. Science. 1974 Jul 26;185(4148):368–370. doi: 10.1126/science.185.4148.368. [DOI] [PubMed] [Google Scholar]
  9. Dvorak A. M., Dvorak H. F., Karnovsky M. J. Uptake of horseradish peroxidase by guinea pig basophilic leukocytes. Lab Invest. 1972 Jan;26(1):27–39. [PubMed] [Google Scholar]
  10. Dvorak A. M., Klebanoff S. J., Henderson W. R., Monahan R. A., Pyne K., Galli S. J. Vesicular uptake of eosinophil peroxidase by guinea pig basophils and by cloned mouse mast cells and granule-containing lymphoid cells. Am J Pathol. 1985 Mar;118(3):425–438. [PMC free article] [PubMed] [Google Scholar]
  11. Ganguly P. Isolation and some properties of fibrinogen from human blood platelets. J Biol Chem. 1972 Mar 25;247(6):1809–1816. [PubMed] [Google Scholar]
  12. George J. N., Nurden A. T., Phillips D. R. Molecular defects in interactions of platelets with the vessel wall. N Engl J Med. 1984 Oct 25;311(17):1084–1098. doi: 10.1056/NEJM198410253111705. [DOI] [PubMed] [Google Scholar]
  13. George J. N., Saucerman S., Levine S. P., Knieriem L. K., Bainton D. F. Immunoglobulin G is a platelet alpha granule-secreted protein. J Clin Invest. 1985 Nov;76(5):2020–2025. doi: 10.1172/JCI112203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. George J. N., Saucerman S. Platelet IgG, IgA, IgM, and albumin: correlation of platelet and plasma concentrations in normal subjects and in patients with ITP or dysproteinemia. Blood. 1988 Jul;72(1):362–365. [PubMed] [Google Scholar]
  15. Handagama P. J., Bainton D. F. Incorporation of a circulating protein into alpha granules of megakaryocytes. Blood Cells. 1989;15(1):59–72. [PubMed] [Google Scholar]
  16. Handagama P. J., George J. N., Shuman M. A., McEver R. P., Bainton D. F. Incorporation of a circulating protein into megakaryocyte and platelet granules. Proc Natl Acad Sci U S A. 1987 Feb;84(3):861–865. doi: 10.1073/pnas.84.3.861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. James H. L., Ganguly P. Identity of human platelet fibrinogen. Biochem Biophys Res Commun. 1975 Apr 7;63(3):659–662. doi: 10.1016/s0006-291x(75)80434-7. [DOI] [PubMed] [Google Scholar]
  18. James H. L., Ganguly P., Jackson C. W. Characterization and origin of fibrinogen in blood platelets. A review with recent data. Thromb Haemost. 1977 Dec 15;38(4):939–954. [PubMed] [Google Scholar]
  19. Karpatkin M., Howard L., Karpatkin S. Studies of the origin of platelet-associated fibrinogen. J Lab Clin Med. 1984 Aug;104(2):223–237. [PubMed] [Google Scholar]
  20. Koch-Weser J., Sellers E. M. Drug therapy. Binding of drugs to serum albumin (second of two parts). N Engl J Med. 1976 Mar 4;294(10):526–531. doi: 10.1056/NEJM197603042941005. [DOI] [PubMed] [Google Scholar]
  21. Kunicki T. J., Newman P. J., Amrani D. L., Mosesson M. W. Human platelet fibrinogen: purification and hemostatic properties. Blood. 1985 Oct;66(4):808–815. [PubMed] [Google Scholar]
  22. Leary J. J., Brigati D. J., Ward D. C. Rapid and sensitive colorimetric method for visualizing biotin-labeled DNA probes hybridized to DNA or RNA immobilized on nitrocellulose: Bio-blots. Proc Natl Acad Sci U S A. 1983 Jul;80(13):4045–4049. doi: 10.1073/pnas.80.13.4045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Leporrier M., Dighiero G., Auzemery M., Binet J. L. Detection and quantification of platelet-bound antibodies with immunoperoxidase. Br J Haematol. 1979 Aug;42(4):605–611. doi: 10.1111/j.1365-2141.1979.tb01173.x. [DOI] [PubMed] [Google Scholar]
  24. Leven R. M., Schick P. K., Budzynski A. Z. Fibrinogen biosynthesis in isolated guinea pig megakaryocytes. Blood. 1985 Feb;65(2):501–504. [PubMed] [Google Scholar]
  25. Lichtman M. A., Chamberlain J. K., Simon W., Santillo P. A. Parasinusoidal location of megakaryocytes in marrow: a determinant of platelet release. Am J Hematol. 1978;4(4):303–312. doi: 10.1002/ajh.2830040402. [DOI] [PubMed] [Google Scholar]
  26. LoBuglio A. F., Court W. S., Vinocur L., Maglott G., Shaw G. M. Immune thrombocytopenic purpura. Use of a 125I-labeled antihuman IgG monoclonal antibody to quantify platelet-bound IgG. N Engl J Med. 1983 Aug 25;309(8):459–463. doi: 10.1056/NEJM198308253090804. [DOI] [PubMed] [Google Scholar]
  27. Milici A. J., Watrous N. E., Stukenbrok H., Palade G. E. Transcytosis of albumin in capillary endothelium. J Cell Biol. 1987 Dec;105(6 Pt 1):2603–2612. doi: 10.1083/jcb.105.6.2603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Moore A., Ross G. D., Nachman R. L. Interaction of platelet membrane receptors with von Willebrand factor, ristocetin, and the Fc region of immunoglobulin G. J Clin Invest. 1978 Nov;62(5):1053–1060. doi: 10.1172/JCI109210. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. NACHMAN R. L. IMMUNOLOGIC STUDIES OF PLATELET PROTEIN. Blood. 1965 May;25:703–711. [PubMed] [Google Scholar]
  30. Nachman R., Levine R., Jaffe E. A. Synthesis of factor VIII antigen by cultured guinea pig megakaryocytes. J Clin Invest. 1977 Oct;60(4):914–921. doi: 10.1172/JCI108846. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pham T. D., Kaplan K. L., Butler V. P., Jr Immunoelectron microscopic localization of platelet factor 4 and fibrinogen in the granules of human platelets. J Histochem Cytochem. 1983 Jul;31(7):905–910. doi: 10.1177/31.7.6343481. [DOI] [PubMed] [Google Scholar]
  32. Plow E. F., Edgington T. S. Unique immunochemical features and intracellular stability of platelet fibrinogen. Thromb Res. 1975 Nov;7(5):729–742. doi: 10.1016/0049-3848(75)90198-x. [DOI] [PubMed] [Google Scholar]
  33. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. ROTHSCHILD M. A., BAUMAN A., YALOW R. S., BERSON S. A. Tissue distribution of I131 labeled human serum albumin following intravenous administration. J Clin Invest. 1955 Sep;34(9):1354–1358. doi: 10.1172/JCI103183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rabellino E. M., Levene R. B., Leung L. L., Nachman R. L. Human megakaryocytes. II. Expression of platelet proteins in early marrow megakaryocytes. J Exp Med. 1981 Jul 1;154(1):88–100. doi: 10.1084/jem.154.1.88. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ryo R., Nakeff A., Huang S. S., Ginsberg M., Deuel T. F. New synthesis of a platelet-specific protein: platelet factor 4 synthesis in a megakaryocyte-enriched rabbit bone marrow culture system. J Cell Biol. 1983 Feb;96(2):515–520. doi: 10.1083/jcb.96.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sander H. J., Slot J. W., Bouma B. N., Bolhuis P. A., Pepper D. S., Sixma J. J. Immunocytochemical localization of fibrinogen, platelet factor 4, and beta thromboglobulin in thin frozen sections of human blood platelets. J Clin Invest. 1983 Oct;72(4):1277–1287. doi: 10.1172/JCI111084. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Shepard J. M., Goderie S. K., Brzyski N., Del Vecchio P. J., Malik A. B., Kimelberg H. K. Effects of alterations in endothelial cell volume on transendothelial albumin permeability. J Cell Physiol. 1987 Nov;133(2):389–394. doi: 10.1002/jcp.1041330226. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Solum N. O., Lopaciuk S. Bovine platelet proteins. 3. Some properties of platelet fibrinogen. Thromb Diath Haemorrh. 1969 Jun 15;21(3):428–440. [PubMed] [Google Scholar]
  41. Steinman R. M., Mellman I. S., Muller W. A., Cohn Z. A. Endocytosis and the recycling of plasma membrane. J Cell Biol. 1983 Jan;96(1):1–27. doi: 10.1083/jcb.96.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Stenberg P. E., Shuman M. A., Levine S. P., Bainton D. F. Redistribution of alpha-granules and their contents in thrombin-stimulated platelets. J Cell Biol. 1984 Feb;98(2):748–760. doi: 10.1083/jcb.98.2.748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Takeda Y. Studies of the metabolism and distribution of fibrinogen in healthy men with autologous 125-I-labeled fibrinogen. J Clin Invest. 1966 Jan;45(1):103–111. doi: 10.1172/JCI105314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Uzan G., Courtois G., Stanckovic Z., Crabtree G. R., Marguerie G. Expression of the fibrinogen genes in rat megakaryocytes. Biochem Biophys Res Commun. 1986 Oct 30;140(2):543–549. doi: 10.1016/0006-291x(86)90766-7. [DOI] [PubMed] [Google Scholar]
  45. Waldmann T. A., Strober W. Metabolism of immunoglobulins. Prog Allergy. 1969;13:1–110. doi: 10.1159/000385919. [DOI] [PubMed] [Google Scholar]
  46. Weisiger R., Gollan J., Ockner R. Receptor for albumin on the liver cell surface may mediate uptake of fatty acids and other albumin-bound substances. Science. 1981 Mar 6;211(4486):1048–1051. doi: 10.1126/science.6258226. [DOI] [PubMed] [Google Scholar]
  47. 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]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES