Download PDF
Thromb Haemost 1982; 48(01): 067-071
DOI: 10.1055/s-0038-1657218
Original Article
Schattauer GmbH Stuttgart

Degranulation of Rabbit Platelets with PAF-Acether: A New Procedure for Unravelling the Mode of Action of Platelet-Activating Substances

B Boris Vargaftig
1   The Unité des Venins, Département de Physiopathologie expérimentale, Institut Pasteur, Paris, France
,
Danielel Joseph
1   The Unité des Venins, Département de Physiopathologie expérimentale, Institut Pasteur, Paris, France
,
Guy Marlas
1   The Unité des Venins, Département de Physiopathologie expérimentale, Institut Pasteur, Paris, France
,
L-G Chevance
2   Station de Microscopie électronique, Département de Physiopathologie expérimentale, Institut Pasteur, Paris, France
› Author Affiliations
Further Information

Publication History

Received 08 March 1982

Accepted 27 May 1982

Publication Date:
13 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

Aggregation and secretion of ATP induced by thrombin, collagen, the snake venom component convulxin and platelet-activating factor (PAF-acether) were studied after the exposure of rabbit platelets to 1 μM of PAF-acether. This concentration, which is around 6 orders of magnitude above the concentration needed to induce full aggregation, was required to remove most of the releasable ATP from the platelets. The depleted platelets aggregated to PAF-acether, to thrombin and to convulxin under conditions where only very low amounts of ATP were secreted, confirming that these agents do not require the release of dense body components to trigger aggregation. Furthermore, when exposure to PAF-acether was associated to inactivation of platelet cyclooxygenase with aspirin, aggregation to thrombin persisted, validating the claim that thrombin induces aggregation by a third pathway unrelated to ADP and to thromboxane A2. Aggregation by collagen was markedly reduced by exposure of the platelets to PAF-acether or to aspirin; when both procedures were associated, aggregation was suppressed. Failure to desensitize the rabbit platelets to PAF-acether upon exposure to high amounts of it indicates the absence of irreversible membrane changes due to PAF-acether, and allows its use as a depleting procedure for the dense body materials, which does not affect platelet membrane components as is the case for thrombin.

Keywords

Aggregation - Aspirin - ATP - Collagen - Convulxin - Platelet-activating factor (PAF-acether) - Thrombin
 

  • References

  • 1 Charo IF, Feinman RD, Detwiller TC, Smith JB, Ingerman CM, Silver MJ. Prostaglandin endoperoxides and thromboxane A2 can induce platelet aggregation in the absence of secretion. Nature 1977; 269: 66-69
    CrossrefPubMedGoogle Scholar
  • 2 Kinlough-Rathbone RL, Reimers HJ, Mustard JF, Packham MA. Sodium arachidonate can induce shape change and aggregation which are independent of the release reaction. Science 1976; 192: 1011-1012
    CrossrefPubMedGoogle Scholar
  • 3 Kinlough-Rathbone RL, Packham MA, Reimers HJ, Cazenave JP, Mustard JF. Mechanisms of platelet shape change, aggregation and release induced by collagen, thrombin, or A23, 187. J Lab Clin Med 1977; 90: 707-719
    PubMedGoogle Scholar
  • 4 Detwiller TC. Hypothetical models for the thrombin-platelet interaction. Ann N Y Acad Sci 1981; 370: 67-71
    CrossrefPubMedGoogle Scholar
  • 5 Godfroid JJ, Heymans F, Michel E, Redeuilh C, Steiner E, Benveniste J. Platelet-activating factor (PAF-acether): total synthesis of 1-0-octadecyl 2-0-acetyl-sn-glycero-3-phosphorylcholine. FEBS Lett 1980; 116: 161-164
    CrossrefPubMedGoogle Scholar
  • 6 Vargaftig BB, Prado-Franceschi J, Chignard M, Lefort J, Marlas G. Activation of guinea-pig platelets induced by convulxin, a substance extracted from the venom of Crotalus durissus cascavella . Europ J Pharmacol 1980; 68: 451-464
    CrossrefPubMedGoogle Scholar
  • 7 Marlas G. Purification and preliminary structure of a potent platelet-aggregating glycoprotein isolated from the venom of Crotalus durissus cascavella . Toxicon 1982; 20: 289-290
    CrossrefPubMedGoogle Scholar
  • 8 Vargaftig BB. Platelet-activation by non-coagulant snake venom components. Toxicon 1982; 20: 279-287
    CrossrefPubMedGoogle Scholar
  • 9 Chignard M, LeCouedic JP, Vargaftig BB, Benveniste J. Platelet-activating factor (PAF-acether) secretion from platelets: effect of aggregating agents. Br J Haematol 1980; 46: 455-464
    CrossrefPubMedGoogle Scholar
  • 10 Reimers HJ, Kinlough-Rathbone RL, Cazenave JP, Senyi AF, Hirsh J, Packham A, Mustard JF. In Vitro and in Vivo Functions of Thrombin-Treated Platelets. Thromb Haemostas 1976; 35: 151-165
    PubMedGoogle Scholar
  • 11 Vargaftig BB. Alternative pathways for platelet aggregation. In Yoshida H, Hagihara Y, Ebashi E. (eds.). Advances in Pharmacology and Therapeutics II . Vol. 6. Pergamon Press; Oxford: 1982: 49
    Google Scholar
  • 12 Cazenave JP, Benveniste J, Mustard JF. Aggregation of Rabbit Platelets by Platelet-Activating Factor Is Independent of the Release Reaction and the Arachidonate Pathway and Inhibited by Membrane-Active Drugs. Lab Invest 1979; 41: 275-285
    PubMedGoogle Scholar
  • 13 Lalau-Keraly C, Benveniste J. Specific desensitization of rabbit platelets by platelet-activating factor (PAF-acether) and derivatives. Br J Haematol 1982; 51: 313-322
    CrossrefPubMedGoogle Scholar
  • 14 Malmsten C, Hamberg M, Svensson J, Samuelsson B. Physiological role of an endoperoxide in human platelets: hemostatic defect due to platelet cyclooxygenase deficiency. Proc Natl Acad Sci USA 1975; 72: 1446-1450
    CrossrefPubMedGoogle Scholar
  • 15 Lagarde M, Byron PA, Vargaftig BB, Dechavanne M. Impairment of platelet thromboxane generation and of the platelet release reaction in two patients with congenital deficiency of platelet cyclo-oxygenase. Br J Haematol 1978; 38: 251-266
    CrossrefPubMedGoogle Scholar