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Thromb Haemost 1988; 60(02): 209-216
DOI: 10.1055/s-0038-1647031
Original Article
Schattauer GmbH Stuttgart

Conditions Affecting the Responses of Human Platelets to Epinephrine

Chantal Lalau Keraly
The Dept, of Pathology, McMaster University, Hamilton, Ontario, Canada and the Dept, of Biochemistry, University of Toronto, Toronto, Ontario, Canada
,
Raelene L Kinlough-Rathbone
The Dept, of Pathology, McMaster University, Hamilton, Ontario, Canada and the Dept, of Biochemistry, University of Toronto, Toronto, Ontario, Canada
,
Marian A Packham
The Dept, of Pathology, McMaster University, Hamilton, Ontario, Canada and the Dept, of Biochemistry, University of Toronto, Toronto, Ontario, Canada
,
Hidenori Suzuki
The Dept, of Pathology, McMaster University, Hamilton, Ontario, Canada and the Dept, of Biochemistry, University of Toronto, Toronto, Ontario, Canada
,
J Fraser Mustard
The Dept, of Pathology, McMaster University, Hamilton, Ontario, Canada and the Dept, of Biochemistry, University of Toronto, Toronto, Ontario, Canada
› Author Affiliations
Further Information

Publication History

Received 22 September 1987

Accepted after revision 20 May 1988

Publication Date:
28 June 2018 (online)

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Summary

Conditions affecting the responses of human platelets to epinephrine were examined. In platelet-rich plasma prepared from blood anticoagulated with hirudin or PPACK (D-pheny- lalanyl-L-prolyl-L-arginine chloromethyl ketone), epinephrine did not cause shape change or aggregation. In a Tyrode-albumin- apyrase solution containing a concentration of Ca2+ in the physiological range, and fibrinogen, epinephrine in concentrations as high as 40 μM did not induce platelet shape change, caused either no primary aggregation or very slight primary aggregation, and did not induce thromboxane formation, release of dense granule contents, or secondary aggregation. In contrast, in citrated platelet-rich plasma, epinephrine induced two phases of aggregation. This is not attributable to the generation of traces of thrombin since the same effects were evident when blood was taken into a combined citrate-hirudin anticoagulant or a combined citrate-PPACK anticoagulant. In a modified Tyrode-albu- min-apyrase solution containing approximately 20 μM Ca2+, 1 mM Mg2+, and fibrinogen, epinephrine induced extensive aggregation after a lag phase, but no primary phase was evident; thromboxane formation and release of dense granule contents accompanied the aggregation response. These responses were also observed when PPACK was included with the acid-citrate- dextrose anticoagulant, and in the washing and resuspending fluids. In the presence of aspirin or the thromboxane receptor blocker BM 13.177 a few small aggregates were detected by particle counting and by scanning electron microscopy; with the latter inhibitor, the platelets in the aggregates retained their disc shape; secondary aggregation and the responses associated with it did not occur. Thus thromboxane A2 formation is not necessary for the formation of these small aggregates, but is required for extensive aggregation and release. As with other weak agonists, the close platelet-to-platelet contact in the low Ca2+ medium appears to be necessary for full secondary aggregation. Omission of fibrinogen from the low Ca2+ medium prevented both primary and secondary aggregation in response to epinephrine. An antibody (10E5) to the glycoprotein Ilb/IIIa complex was completely inhibitory in the presence of fibrinogen. Thus the response of human platelets to epinephrine is influenced by the concentration of Ca2+ and the presence of fibrinogen in the medium in which they are suspended.

Keywords

Epinephrine - Human platelets - Scanning electron microscopy - Calcium - Fibrinogen - Platelet aggregation
 

  • References

  • 1 Mustard JF, Perry DW, Kinlough-Rathbone RL, Packham MA. Factors responsible for ADP-induced release reaction of human platelets. Am J Physiol 1975; 228: 1757-1765
    PubMedGoogle Scholar
  • 2 Macfarlane DE, Walsh PN, Mills DCB, Holmsen H, Day HJ. The role of thrombin in ADP-induced platelet aggregation and release. A critical evaluation. Br J Haematol 1975; 30: 457-463
    CrossrefPubMedGoogle Scholar
  • 3 Glusa E, Markwardt F. Adrenaline-induced reactions of human platelets in hirudin plasma. Haemostasis 1980; 9: 188-192
    PubMedGoogle Scholar
  • 4 Lages B, Weiss HJ. Dependence of human platelet functional responses on divalent cations: aggregation and secretion in heparin- and hirudin-anticoagulated platelet-rich plasma and the effects of chelating agents. Thromb Haemostas 1981; 45: 173-179
    PubMedGoogle Scholar
  • 5 Kinlough-Rathbone RL, Mustard JF, Packham MA, Perry DW, Reimers HJ, Cazenave JP. Properties of washed human platelets. Thromb Haemostas 1977; 37: 291-308
    PubMedGoogle Scholar
  • 6 Massini P. The role of calcium in the stimulation of platelets. In: Platelets and Thrombosis. Mills DC B, Pareti FI. (eds) Academic Press; London : 1977. pp 33-43
    Google Scholar
  • 7 Cazenave JP, Sutter A, Hemmendinger S, Wiesel ML, Lanza F, Daver J. Adrenaline activates human platelets but does not cause primary aggregation if thrombin generation is inhibited by hirudin. Thromb Haemostas 1981; 46: 95 (Abstr).
    PubMedGoogle Scholar
  • 8 Lanza F, Beretz A, Stierle A, Hanau D, Kubina M, Cazenave JP. Adrenaline potentiates human platelet activation but is not an aggregating agent. Am J Physiol (In press).
    Google Scholar
  • 9 O’Brien JR. Some effects of adrenaline and antiadrenaline compounds on platelets in vitro and in vivo. Nature (London) 1963; 200: 763-764
    CrossrefPubMedGoogle Scholar
  • 10 Haslam RJ. Mechanisms of blood platelet aggregation.. In: Physiology of Hemostasis and Thrombosis. Johnson SA, Seegers WH. (eds) Thomas; Springfield: 1967. pp 88-112
    Google Scholar
  • 11 Figures WR, Scearce LM, Wachtfogel Y, Chen J, Colman RF, Colman RW. Platelet ADP receptor and a2-adrenoreceptor interaction. Evidence for an ADP requirement for epinephrine-induced platelet activation and an influence of epinephrine on ADP binding. J Biol Chem 1986; 261: 5981-5986
    PubMedGoogle Scholar
  • 12 Plow EF, Marguerie GA. Induction of the fibrinogen receptor on human platelets by epinephrine and the combination of epinephrine and ADP. J Biol Chem 1980; 255: 10971-10977
    PubMedGoogle Scholar
  • 13 Bennett JS, Vilaire G. Exposure of platelet fibrinogen receptors by ADP and epinephrine. J Clin Invest 1979; 64: 1393-1401
    CrossrefPubMedGoogle Scholar
  • 14 Di MinnoG, Capitanio AM, Thiagarajan P, Martinez J, Murphy S. Exposure of fibrinogen receptors on fresh and stored platelets by ADP and epinephrine as single agents and as a pair. Blood 1983; 61: 1054-1059
    PubMedGoogle Scholar
  • 15 Peerschke EI. Induction of human platelet fibrinogen receptors by epinephrine in the absence of released ADP. Blood 1982; 60: 71-77
    PubMedGoogle Scholar
  • 16 Mills DC B. Changes in the adenylate energy charge in human blood platelets induced by adenosine diphosphate. Nature New Biol 1973; 243: 220-222
    CrossrefPubMedGoogle Scholar
  • 17 Mustard JF, Packham MA. Factors influencing platelet function: adhesion, release, and aggregation. Pharmacol Rev 1970; 22: 97-187
    PubMedGoogle Scholar
  • 18 Milton JG, Frojmovic MM. Adrenaline and adenosine-diphosphate- induced platelet aggregation require shape change. Importance of pseudopods. J Lab Clin Med 1984; 104: 805-815
    PubMedGoogle Scholar
  • 19 O’Brien JR, Woodhouse MA. Platelets: Their size, shape and stickiness in vitro: Degranulation and propinquity. Exp Biol Med 1968; 3: 99-102
    PubMedGoogle Scholar
  • 20 Mills DC B, Robb IA, Roberts GC K. The release of nucleotides, 5- hydroxytryptamine and enzymes from human blood platelets. J Physiol 1968; 195: 715-729
    CrossrefPubMedGoogle Scholar
  • 21 Gerrard JM, White JG. The influence of aspirin and indomethacin on the platelet contractile wave. Am J Pathol 1976; 82: 513-526
    PubMedGoogle Scholar
  • 22 O’Brien JR. A comparison of platelet aggregation produced by seven compounds and a comparison of their inhibitors. J Clin Pathol 1964; 17: 275-281
    CrossrefPubMedGoogle Scholar
  • 23 Ardlie NG, Glew G, Schwartz CJ. Influence of catecholamines on nucleotide induced platelet aggregation. Nature 1966; 212: 415-417
    CrossrefPubMedGoogle Scholar
  • 24 Mills DC B, Roberts GC K. Effects of adrenaline on human blood platelets. J Physiol 1967; 193: 443-453
    CrossrefPubMedGoogle Scholar
  • 25 Grant JA, Scrutton MC. Positive interaction between agonists in the aggregation of human blood platelets: interaction between ADP, adrenaline and vasopressin. Br J Haematol 1980; 44: 109-125
    CrossrefPubMedGoogle Scholar
  • 26 Huang EM, Detwiler TC. Characteristics of platelet agonists. Blood 1981; 57: 685-691
    PubMedGoogle Scholar
  • 27 Kettner C, Shaw E. D-Phe-Pro-ArgCH2Cl - A selective affinity label for thrombin. Thromb Res 1979; 14: 969-973
    CrossrefPubMedGoogle Scholar
  • 28 Kinlough-Rathbone RL, Packham MA, Mustard JF. Platelet aggregation.. In: Methods in Hematology. Measurements of Platelet Function Harker LA, Zimmerman TS. (eds) Churchill Livingstone; Edinburgh : 1983. pp 64-91
    Google Scholar
  • 29 Molnar J, Lorand L. Studies on apyrases. Arch Biochem Biophys 1961; 93: 353-363
    CrossrefPubMedGoogle Scholar
  • 30 Lawrie JS, Ross J, Kemp GD. Purification of fibrinogen and the separation of its degradation products in the presence of calcium ions. Biochem Soc Trans 1979; 7: 693-694
    CrossrefPubMedGoogle Scholar
  • 31 Davies JA, Essien E, Cazenave JP, Kinlough-Rathbone RL, Gent M, Mustard JF. The influence of red blood cells on the effects of aspirin or sulphinpyrazone on platelet adherence to damaged rabbit aorta. Br J Haematol 1979; 42: 283-291
    CrossrefPubMedGoogle Scholar
  • 32 Mustard JF, Perry DW, Ardlie NG, Packham MA. Preparation of suspensions of washed platelets from humans. Br J Haematol 1972; 22: 193-204
    CrossrefPubMedGoogle Scholar
  • 33 Greenberg J, Packham MA, Cazenave JP, Reimers HJ, Mustard JF. Effects on platelet function of removal of platelet sialic acid by neuraminidase. Lab Invest 1975; 32: 476-484
    PubMedGoogle Scholar
  • 34 Packham MA, Guccione MA, Greenberg JP, Kinlough-Rathbone RL, Mustard JF. Release of 14C-serotonin during initial platelet changes induced by thrombin, collagen, or A 23187. Blood 1977; 50: 915-926
    PubMedGoogle Scholar
  • 35 Packham MA, Kinlough-Rathbone RL, Mustard JF. Thromboxane A2 causes feedback amplification involving extensive thromboxane A 2formation upon close contact of human platelets in media with a low concentration of ionized calcium. Blood 1987; 70: 647-651
    PubMedGoogle Scholar
  • 36 O’Brien JR. Aspirin and platelet aggregation. Lancet 1968; 1: 204-205
    PubMedGoogle Scholar
  • 37 Weiss HJ, Aledort LM, Kochiva S. The effect of salicylates on the hemostatic properties of platelets in man. J Clin Invest 1968; 47: 2169-2180
    CrossrefPubMedGoogle Scholar
  • 38 Zucker MB, Peterson J. Effect of acetylsalicylic acid, other nonsteroidal anti-inflammatory agents, and dipyridamole on human blood platelets. J Lab Clin Med 1970; 76: 66-75
    PubMedGoogle Scholar
  • 39 Best LC, Holland TK, Jones PB B, Russell RG G. The interrelationship between thromboxane biosynthesis, aggregation and 5-hydroxytryptamine secretion in human platelets in vitro. Thromb Haemostas 1980; 43: 38-40
    PubMedGoogle Scholar
  • 40 Patscheke H, Stegmeier K. Investigations on a selective non-prostanoic thromboxane antagonist, BM 13.177, in human platelets. Thromb Res 1984; 33: 277-288
    CrossrefPubMedGoogle Scholar
  • 41 Huang EM, Detwiler TC. Stimulus-response coupling mechanisms.. In: Biochemistry of Platelets. Phillips DR, Shuman MA. (eds). Academic Press; New York : 1986. pp 1-67
    Google Scholar
  • 42 Bull BS, Zucker MB. Changes in platelet volume produced by temperature, metabolic inhibitors, and aggregating agents. Proc Soc Exp Biol Med 1965; 120: 296-301
    CrossrefPubMedGoogle Scholar
  • 43 Oliver JA, Albrecht RM. Colloidal gold labelling of fibrinogen receptors in epinephrine- and ADP-activated platelet suspensions. Scanning Microscopy 1987; 1: 745-756
    PubMedGoogle Scholar
  • 44 Rao GHR, Johnson GJ, White JG. Influence of epinephrine on the aggregation response of aspirin treated platelets. Prostaglandins Med 1980; 5: 45-58
    CrossrefPubMedGoogle Scholar
  • 45 Cameron HA, Ardlie NG. The facilitating effects of adrenaline on platelet aggregation. Prostaglandins Leukotrienes Med 1982; 9: 117-128
    CrossrefPubMedGoogle Scholar
  • 46 Peerschke EIB. Effect of epinephrine on fibrinogen receptor exposure by aspirin-treated platelets and platelets from concentrates in response to ADP and thrombin. Am J Hematol 1984; 16: 335-445
    CrossrefPubMedGoogle Scholar
  • 47 Mustard JF, Packham MA, Kinlough-Rathbone RL, Perry DW, Regoeczi E. Fibrinogen and ADP-induced platelet aggregation. Blood 1978; 52: 453-466
    PubMedGoogle Scholar
  • 48 Cattaneo M, Kinlough-Rathbone RL, Lecchi A, Bevilacqua C, Packham MA, Mustard JF. Fibrinogen-independent aggregation and deaggregation of human platelets: studies in two afibrinogenemic patients. Blood 1987; 70: 221-226
    PubMedGoogle Scholar
  • 49 Scrutton MC, Wallis RB. Catecholamine receptors.. In: Platelets in Biology and Pathology (Vol.; 2). Gordon JL. (ed) Elsevier/North-Holland Biomedical Press; Amsterdam : 1981. pp 179-210
    Google Scholar
  • 50 Ardlie NG, McGuiness JA, Garrett JJ. Effect on human platelets of catecholamines at levels achieved in the circulation. Atherosclerosis 1985; 58: 251-259
    CrossrefPubMedGoogle Scholar