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Thromb Haemost 1985; 54(03): 574-578
DOI: 10.1055/s-0038-1660073
Original Article
Schattauer GmbH Stuttgart

Freeze-Fracture Studies of Human Blood Platelets Activated by Thrombin Using Rapid Freezing

Hans Hols
1   The Department of Molecular Cell Biology, State University Utrecht, The Netherlands
,
Jan J Sixma
2   The Department of Haematology, State University Utrecht, The Netherlands
,
J Leunissen-Bijvelt
1   The Department of Molecular Cell Biology, State University Utrecht, The Netherlands
,
Arie Verkley
3   The Institute of Molecular Biology, State University Utrecht, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 02 February 1985

Accepted 27 June 1985

Publication Date:
19 July 2018 (online)

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Summary

In this study the influence of thrombin activation on human blood platelets has been followed by freeze-fracturing electron microscopy using rapid freezing in order to catch the initial changes in shape and the morphological alterations during the process of exocytosis of secretory granules. We found that isolation of the platelets by itself leads to some degree of shape change, which made it impossible to study the resting discoid platelet by rapid freezing.

Activation of the platelets by thrombin induced dilation of the “surface connecting system (SCS)” with formation of large vacuoles as a result of fusion of the secretory granules with SCS. No intermediary fusion stages or structures were observed even using rapid freezing. Volcano-like protrusions and the corresponding complementary pits were seen at the SCS. These structures were interpreted by us as fractures through protoplasmic channels crossing the SCS. These channels originate during the swelling of the SCS as a result of the fusion of secretory granules with the SCS.

Keywords

Blood platelets - Freeze - fracture - Secretion
 

  • References

  • 1 White JG. Electron microscopic studies of platelet secretion. In: Progress in Hemostasis and Thrombosis. Spaet T. (ed) Grune and Stratton; New York: 1974: 49-99
    Search in Google Scholar
  • 2 Behnke O, Kristensen BI, Engdahl Nielsen L. Electron-microscopical observations on actinoid and myosinoid filament in blood platelets. J Ultrastr Res 1971; 37: 351-369
    CrossrefPubMedSearch in Google Scholar
  • 3 Nachmias V, Gullander J, Asch A. Shape and cytoplasmic filaments ir control and lidocaine-treated human platelets. Blood 1977; 50: 39-53
    PubMedSearch in Google Scholar
  • 4 Behnke O. The morphology of platelet membrane system. Sei Haematol 1970; 3: 3
    PubMedSearch in Google Scholar
  • 5 Tavassoli M. Megakaryocyt-platelet axis and the process of platelei formation and release. Blood 1980; 55: 537-545
    PubMedSearch in Google Scholar
  • 5 White JG. Interaction of membrane systems in blood platelets. Am J Pathol 1972; 66: 295
    PubMedSearch in Google Scholar
  • 6 Gerrard JM, White JM, Rao G HR. Effects of the ionophore A2318' on blood platelets. II. Influence on ultrastructure. Am J Pathol 1974; 77: 151-166
    PubMedSearch in Google Scholar
  • 7 Gerrard JM, White JG, Rao G HR, Townsend D. Localization o: platelet prostaglandin production in the platelet dense tubular system. Am J Pathol 1976; 83: 283-298
    PubMedSearch in Google Scholar
  • 8 Behnke O. Electron microscopic observations of the membrane systems of the rat platelets. Anat Rec 1967; 158: 121-138
    CrossrefPubMedSearch in Google Scholar
  • 9 Behnke O. Electron microscopical observation on the surface coating of human blood platelets. J Ultrastruct Res 1968; 24: 51
    CrossrefPubMedSearch in Google Scholar
  • 10 Behnke O. An electron microscope study of the megakaryocyte of the rat marrow. The development of the demarcation system and the platelet surface coat. J Ultrastruc Res 1968; 24: 412
    CrossrefPubMedSearch in Google Scholar
  • 11 Shaklai M, Tavassoli M. Demarcation membrane system in ra megakaryocyte and the mechanism of platelet formation: A mem brane reorganization process. J Ultrastruct Res 1978; 62: 270-285
    CrossrefPubMedSearch in Google Scholar
  • 12 Ruska C, Schulz H. Elektronenmikroskopische Darstellung vor Thrombozyten mit der Gefrieratztechnik. Klin Wschr 1968; 46: 689-696
    CrossrefPubMedSearch in Google Scholar
  • 13 Hoak JC. Freeze-etching studies of human platelets. Blood 1972; 40: 514-522
    PubMedSearch in Google Scholar
  • 14 Reddick RL, Mason RG. Freeze etch observation on the plasim membrane and other structures of normal and abnormal platelets. An J Pathol 1973; 70: 437-488
    PubMedSearch in Google Scholar
  • 15 Chevalier J, Nurden AT, Thiery JW, Savariau E, Caen JP. Freeze facture studies on the plasma membranes of normal human, thrombas thenic and Bernard-Soulier platelets. J Lab Clin Med 1970; 94: 232-245
    PubMedSearch in Google Scholar
  • 16 Zucker-Franklin D. Endocytosis by human platelets. Metabolic anc freeze-fracture studies. J Cell Biol 1981; 91: 706-715
    CrossrefPubMedSearch in Google Scholar
  • 17 Bachmann L, Schmitt WW. Weniger Artefacte in der Gefrieratzunj durch erhohte Einfriergeschwindigkeit. Naturwissenschaften 1971; 58: 217-218
    CrossrefPubMedSearch in Google Scholar
  • 18 De Bruyn WC, van Buitenen J MH. X-ray microanalysis of aldehyde fixed glycogen contrast-stained OsVIFeII and OsVIRuIV complexes. Histochem Cytochem 1980; 281: 1242
    PubMedSearch in Google Scholar
  • 19 Massini P, Lüscher EF. The induction of the release reaction ii human blood platelets by close cell contact. Thrombos Diathe Haemorrh 1971; 25: 13-20
    PubMedSearch in Google Scholar
  • 20 Bunting S, Simmons PM, Moncada S. Inhibition of platelet activatioi by prostacyclin. Thromb Res 1981; 21: 89-102
    CrossrefPubMedSearch in Google Scholar
  • 21 Holme R, Sixma JJ, Mürer EH, Hovig T. Demonstration of platele fibrinogen secretion via the surface connecting system. Thromb Re 1973; 3: 347-356
    PubMedSearch in Google Scholar
  • 22 Breton-Gorius J, Vanhaeke D, Tabilio A, Vainchenker W, Deschamps JF, Xu FS. Glycoprotein lb. Identification during norma and pathological megakaryocytic maturation. Blood Cells 1983; 9: 275-291
    PubMedSearch in Google Scholar
  • 23 Verkleij AJ. Lipidic intramembrane particles. Biochim Biophys Act 1984; 779: 43-63
    CrossrefPubMedSearch in Google Scholar
  • 24 Chandler DE, Heuser JE. Arrest of membrane fusion events in mas cells by quick-freezing. J Cell Biol 1980; 86: 666-674
    CrossrefPubMedSearch in Google Scholar
  • 25 Milton JG, Frojmovie MM. Invaginated plasma membrane of humai platelets: evagination and measurement in normal and "giant" cell platelets. J Lab Clin Med 1979; 93: 162-170
    PubMedSearch in Google Scholar