Download PDF
Thromb Haemost 1976; 35(03): 717-736
DOI: 10.1055/s-0038-1647970
Original Article
Schattauer GmbH

Aggregation of Platelets and Inert Particles Induced by Thrombin

Frangis C. Chao
1   Center for Blood Research; Department of Medicine, New England Deaconess Hospital and Harvard Medical School, and Department of Physics, Boston College, Boston, Massachusetts, U.S.A.
,
James L. Tullis
1   Center for Blood Research; Department of Medicine, New England Deaconess Hospital and Harvard Medical School, and Department of Physics, Boston College, Boston, Massachusetts, U.S.A.
,
Gail S. Conneely
1   Center for Blood Research; Department of Medicine, New England Deaconess Hospital and Harvard Medical School, and Department of Physics, Boston College, Boston, Massachusetts, U.S.A.
,
John W. Lawler
1   Center for Blood Research; Department of Medicine, New England Deaconess Hospital and Harvard Medical School, and Department of Physics, Boston College, Boston, Massachusetts, U.S.A.
› Author Affiliations
Further Information

Publication History

Received 10 December 1974

Accepted 20 August 1975

Publication Date:
02 July 2018 (online)

Also available at
    Permissions and Reprints

Summary

Thrombin-induced platelet aggregation and release were investigated in washed platelet suspensions and in suspensions of inert particles in order to evaluate the role of fibrinogen-fi-brin transformation in aggregometer tracings. Thrombin (0.25–2.0 U/ml) produced two waves of light transmission increase (LTI) in both platelet and inert particle suspensions containing fibrinogen, and concomittantly aggregates were observed under phase microscopy. Without fibrinogen, thrombin induced rapid release of platelet ADP but failed to cause second wave of LTI. The kinetics of LTI in platelet and inert particle systems were related to both thrombin and fibrinogen concentrations. A rapid second wave of LTI could be produced by direct interaction of thrombin-treated platelets or inert particles with polymerizing fibrin, and was inhibited by sodium sulfite and low pH of 5.1 which prevent fibrin monomer polymerization. No fibrin strands were noted in platelet aggregates fixed at the completion of the second wave of LTI. Apyrase and PGE1 inhibited the rate of first but not that of second wave LTI. The results suggest that the release of platelet ADP induced by thrombin primarily affects the first phase aggregation, and the second phase may result from interaction of thrombin-exposed platelets and polymerizing fibrin. Thus, the blood coagulation mechanism may be directly involved in platelet aggregation.

 

  • References

  • 1 Ardlie N. G, Han P. 1974; Enzymatic basis for platelet aggregation and release: The significance of the ‘platelet atmosphere’ and the relationship between platelet function and blood coagulation. British Journal of Hematology 26: 331.
    CrossrefPubMedSearch in Google Scholar
  • 2 Ashcroft G, Crawford B, Binns J. K, MacDougall E. J. 1964; Estimation of 5-hydroxytryp-tamine in human blood. Clinica chimica acta 9: 364.
    CrossrefPubMedSearch in Google Scholar
  • 3 Baenziger N. L, Brodie G. N, Majerus P. W. 1971; A thrombinsensitive protein of human platelet membrane. Proceedings of the National Academy of Sciences 68: 240.
    CrossrefPubMedSearch in Google Scholar
  • 4 Boyer M. H, Shainoff J. R, Ratnoff O. D. 1972; Acceleration of fibrin polymerization by calcium ions. Blood 39: 382.
    PubMedSearch in Google Scholar
  • 5 Chao F. C, Tullis J. L. 1973; The in vitro effects of mithramycin on the aggregation and the calcium uptake of human platelets. Thrombosis et Diathesis Haemorrhagica 29: 712.
    PubMedSearch in Google Scholar
  • 6 Chao F. C, Tullis J. L, Kenney D. M, Conneely G. S, Doyle J. R. 1974 Concentration effects of platelets, fibrinogen and thrombin on platelet aggregation and fibrin clotting. Thrombosis et Diathesis Haemorrhagica. 32
    PubMedSearch in Google Scholar
  • 7 Doolittle R. F. 1973. Structural aspects of the fibrinogen to fibrin conversion. In: Anfinsen C. B, Edsall J. T, Richards F. M. (eds.) Advances in Protein Chemistry.. Academic Press; New York: 1.
    Search in Google Scholar
  • 8 Elias T. P, Iyers G. Y. N. 1967; Some inhibitors of the fibrinogen-fibrin conversion. Thrombosis et Diathesis Haemorrhagica 18: 499.
    PubMedSearch in Google Scholar
  • 9 Fang P, Chao F. C, Tullis J. L. 1974; Kinetic analysis of thrombin-induced aggregation of human platelets. I. Thrombin dependency. Microvascular Research 8: 70.
    PubMedSearch in Google Scholar
  • 10 Ganguly P. 1974; Binding of thrombin to human platelets. Nature 247: 306.
    CrossrefPubMedSearch in Google Scholar
  • 11 Glover G, Shaw E. 1971; The purification of thrombin and isolation of a peptide containing the active center histidine. The Journal of Biological Chemistry 246: 4594.
    PubMedSearch in Google Scholar
  • 12 Haslam R. J. 1967. Mechanism of blood platelet aggregation. In: Johnson S. A, Seegers W. H. (eds.) Physiology of Hemostasis and Thrombosis.. Charles C. Thomas; Springfield, Illinois: 88.
    Search in Google Scholar
  • 13 Holmsen H, Holmsen I, Bernhardsen A. 1966; Micro determination of adenosine dephosphate and adenosine triphosphate in plasma with the firefly luciferase system. Analytical Biochemistry 17: 456.
    CrossrefPubMedSearch in Google Scholar
  • 14 Horowitz H. I, Papayoanou M. F. 1968; Activation of platelet factor 3 by adenosine 5’ diphosphate. Thrombosis et Diathesis Haemorrhagica 19: 18.
    PubMedSearch in Google Scholar
  • 15 Hovig T. 1962; The ultrastructure of rabbit blood platelet aggregates. Thrombosis et Diathesis Haemorrhagica 8: 455.
    PubMedSearch in Google Scholar
  • 16 Hugues J. 1959; Agglutination precoce des plaquettes au cours de la formation du clou hemostatique. Thrombosis et Diathesis Haemorrhagica 3: 177.
    PubMedSearch in Google Scholar
  • 17 Johnson S. A. 1971. Platelets in hemostasis and thrombosis. In: Johnson S. A. (ed.) The Circulating Platelet.. Academic Press; New York: 355.
    CrossrefSearch in Google Scholar
  • 18 Klein P. D, Seegers W. E. 1950; The nature of plasma anti-thrombin activity. Blood 5: 742.
    PubMedSearch in Google Scholar
  • 19 Larrieu M. J, Rigollot C, Kubisz P. 1970; Platelet aggregation induced by soluble fibrin monomer complexes. Life Science 9: 1111.
    CrossrefPubMedSearch in Google Scholar
  • 20 Luft J. H. 1961; Improvements in epoxy resin embedding methods. Journal Biophysics Biochemistry Cytology 9: 409.
    PubMedSearch in Google Scholar
  • 21 Macfarlane R. G. 1972. Hemostasis. In: Biggs R. (ed.) Human Blood Coagulation, Hemostasis and Thrombosis.. Blackwell; Oxford: 543.
    Search in Google Scholar
  • 22 Mommaerts W. F. M. 1945; On the nature of forces operating in blood clotting. II. The clotting of fibrinogen as a two-step reaction. Journal of General Physiology 29: 113.
    PubMedSearch in Google Scholar
  • 23 Müller-Berghaus G, Heinrich D. 1972; Fibrin monomer and platelet aggregation in vitro and in vivo . British Journal of Haematology 23: 177.
    PubMedSearch in Google Scholar
  • 24 Mustard J. F, Packham M. A. 1970; Factors influencing platelet function: adhesion, release and aggregation. Pharmacological Review 22: 97.
    PubMedSearch in Google Scholar
  • 25 Niewiarowski S, Regoeczi E, Stewart G. J, Senyi A. F, Mustard J. F. 1973; Platelet interaction with polymerizing fibrin. Journal of Clinical Investigation 51: 685.
    PubMedSearch in Google Scholar
  • 26 Packham M. A, Guccione M. A, Chang P. L, Mustard J. F. 1973; Platelet aggregation and release: effects of low concentrations of thrombin or collagen. American Journal of Physiology 225: 38.
    PubMedSearch in Google Scholar
  • 27 Phillips D. R, Agin P. P. 1974; Thrombin substrates and the proteolytic site of thrombin action on human-platelet plasma membrane. Biochimica et Biophysica Acta 352: 218.
    CrossrefPubMedSearch in Google Scholar
  • 28 Reynolds E. S. 1963; The use of lead citrate at high pH as an electron opaque stain in electron microscopy. Journal of Cell Biology 17: 208.
    CrossrefPubMedSearch in Google Scholar
  • 29 Solum N. O. 1966; Platelet aggregation during fibrin polymerization. Scandinavian Journal of Clinical and Laboratory Investigation 18: 577.
    CrossrefPubMedSearch in Google Scholar
  • 30 Solum N. O, Stormorken H. 1965; Influence of fibrinogen on the aggregation of washed human blood platelets induced by adenosine diphosphate, thrombin, collagen and adrenaline. Scandinavian Journal of Clinical and Laboratory Investigation 17 (suppl 84) 170.
    PubMedSearch in Google Scholar
  • 31 Steiner M. 1973; Effect of thrombin on the platelet membrane. Biochimica et Biophysica Acta 323: 653.
    CrossrefPubMedSearch in Google Scholar
  • 32 Stemerman M. B. 1974. Platelet interaction with intimal connective tissue. In: Baldini M. G, Ebbe S. (eds.) Platelets: Production, Function and Storage.. Grune S. Stratton; New York: 157.
    Search in Google Scholar
  • 33 Tangen O, Berman H. J, Marfey P. 1971; Gel filtration. A new technique for separation of blood platelets from plasma. Thrombosis et Diathesis Haemorrhagica 25: 268.
    PubMedSearch in Google Scholar
  • 34 Thomas D. P. 1967; Effect of catecholamines on platelet aggregation caused by thrombin. Nature 215: 298.
    CrossrefPubMedSearch in Google Scholar
  • 35 Tollefsen D. M, Feagler J. R, Majerus P. W. 1974; The binding of thrombin to the surface of human platelets. The Journal of Biological Chemistry 249: 2646.
    PubMedSearch in Google Scholar
  • 36 Tullis J. L, Tinch R. J, Baudanza P, Gibson J. G, Diforte S, Conneely G, Murthy K. 1971; Plateletpheresis in a disposable system. Transfusion 11: 368.
    CrossrefPubMedSearch in Google Scholar
  • 37 Walsh P. N. 1972; The role of platelets in the contact phase of blood coagulation. British Journal of Haematology 22: 237.
    CrossrefPubMedSearch in Google Scholar
  • 38 Zucker M. B. 1972. Platelet function. In: Williams W. J, Beutler E, Erslev A. J, Rundles R. W. (eds.) Hematology.. McGraw-Hill; New York: 1014.
    Search in Google Scholar