Thromb Haemost 1984; 51(02): 140-144
DOI: 10.1055/s-0038-1661045
Original Article
Schattauer GmbH Stuttgart

Effects of Tris on Responses of Human and Rabbit Platelets to Aggregating Agents

M A Packham
The Department of Biochemistry, University of Toronto, Toronto, and Department of Pathology, McMaster University, Hamilton, Ontario, Canada
,
M A Guccione
The Department of Biochemistry, University of Toronto, Toronto, and Department of Pathology, McMaster University, Hamilton, Ontario, Canada
,
M Nina
The Department of Biochemistry, University of Toronto, Toronto, and Department of Pathology, McMaster University, Hamilton, Ontario, Canada
,
R L Kinlough-Rathbone
The Department of Biochemistry, University of Toronto, Toronto, and Department of Pathology, McMaster University, Hamilton, Ontario, Canada
,
J F Mustard
The Department of Biochemistry, University of Toronto, Toronto, and Department of Pathology, McMaster University, Hamilton, Ontario, Canada
› Author Affiliations
Further Information

Publication History

Received 29 September 1983

Accepted 03 September 1983

Publication Date:
19 July 2018 (online)

Summary

Despite reports that Tris [tris(hydroxymethyl)aminomethane] affects platelets, it is often used to buffer suspending media. Human or rabbit platelets were washed and resuspended in Tyrode solution containing apyrase and 0.35% albumin. Addition of 15 mM Tris partially inhibited primary aggregation induced by 10 μM ADP and inhibited aggregation and release of 14C- serotonin from prelabelled platelets stimulated with low concentrations of thrombin (0.05-0.2 U/mL), or collagen. Platelets resuspended in 15 mM Tris, 0.15 M NaCl, 0.35% albumin, pH 7.5, did not aggregate in response to 10 μM ADP whereas platelets in Tyrode-albumin aggregated extensively. Ca2+ (5 mM) did not overcome the inhibition of thrombin-induced aggregation. Tris (15 or 1.5 mM) potentiated aggregation and release induced by sodium arachidonate (20-50 μM) or the ionophore A23187 (0.6-1 μM). Pretreatment of platelets with aspirin did not prevent potentiation by A23187, indicating that it is not mediated through activation of the arachidonate pathway. The inhibitory and potentiating effects of Tris are similar to those of amino sugars, lysine, arginine and primary amines such as methylamine and cadaverine, and may represent general effects of amines on platelets. Potentiation of the effects of some aggregating agents and inhibition of others re-emphasizes the concept that there are several different mechanisms through which aggregation can occur. Tris-based buffers are unsuitable for platelet suspending media and their use as solvents for aggregating agents or inhibitors should be limited.

 
  • References

  • 1 Haslam RJ, Rosson GM. Aggregation of human blood platelets by vasopressin. Am J Physiol 1972; 223: 958-967
  • 2 Baenziger NL, Majerus PW. Isolation of human platelets and platelet surface membranes. Fleischer S, Packer L. (eds) In Methods in enzymology. New York: Academic Press; 1974. 31 A 149-155
  • 3 Tangen O, Berman HJ, Marfey P. Gel filtration: a new technique for separation of blood platelets from plasma. Thrombos Diathes Haemorrh 1971; 25: 268-278
  • 4 Hawiger J, Hawiger A, Steckley S, Timmons S, Cheng C. Membrane changes in human platelets by lipopolysaccharide endotoxin. Br J Haematol 1977; 35: 285-299
  • 5 Ganguly P, Sonnichsen WJ. Binding of thrombin to human platelets and its possible significance. Br J Haematol 1976; 34: 291-301
  • 6 Zucker WH, Shermer RW, Mason RG. Ultrastructural comparison of human platelets separated from blood by various means. Am J Pathol 1974; 77: 255-262
  • 7 Lages B, Scrutton MC, Holmsen H. Studies on gel-filtered human platelets: isolation and characterization in a medium containing no added Ca2+, Mg2+, or K+ . J Lab Clin Med 1975; 85: 811-825
  • 8 Cazenave J-P, Packham MA, Mustard JF. Adherence of platelets to a collagen-coated surface: development of a quantitative method. J Lab Clin Med 1973; 82: 978-990
  • 9 Kinlough-Rathbone RL, Reimers HJ, Mustard JF, Packham MA. Sodium arachidonate can induce platelet shape change and aggregation which are independent of the release reaction. Science 1976; 192: 1011-1012
  • 10 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
  • 11 Molnar J, Lorand L. Studies on apyrases. Arch Biochem Biophys 1961; 93: 353-363
  • 12 Mustard JF, Perry DW, Ardlie NG, Packham MA. Preparation of suspensions of washed platelets from humans. Br J Haematol 1972; 22: 193-204
  • 13 Ardlie NG, Packham MA, Mustard JF. Adenosine diphosphate- induced platelet aggregation in suspensions of washed rabbit platelets. Br J Haematol 1970; 19: 7-17
  • 14 Ardlie NG, Perry DW, Packham MA, Mustard JF. Influence of apyrase on stability of suspensions of washed rabbit platelets. Proc Soc Exp Biol Med 1971; 136: 1021-1023
  • 15 Harfenist EJ, Packham MA, Mustard JF. Reversibility of the association of fibrinogen with rabbit platelets exposed to ADP. Blood 1980; 56: 189-198
  • 16 Greenberg J, Packham MA, Cazenave J-P, Reimers H-J, Mustard JF. Effects on platelet function of removal of platelet sialic acid by neuraminidase. Lab Invest 1975; 32: 476-484
  • 17 Ganguly P, Bradford HR. Inhibition of platelet aggregation by primary amines. Evidence for a possible role of membrane-associated calcium Biochim Biophys Acta 1982; 714: 192-199
  • 18 Gartner TK, Williams DC, Minion FC, Phillips DR. Thrombin- induced platelet aggregation is mediated by a platelet plasma membrane-bound lectin. Science 1978; 200: 1281-1283
  • 19 Kinlough-Rathbone RL, Chahil A, Perry DW, Packham MA, Mustard JF. Effect of amine sugars that block platelet lectin activity on fibrinogen binding to washed rabbit or human platelets. Blood 1979; 54 Suppl (Suppl. 01) 249 a
  • 20 Nachman RL, Leung LL. Complex formation of platelet membrane glycoproteins IIb and IIIa with fibrinogen. J Clin Invest 1982; 69: 263-269
  • 21 Houston DS, Gerrard JM, McCrea JM, Glover S, Butler AM. The influence of amines on various platelet functions. Biochim Biophys Acta 1983; 734: 267-273
  • 22 Ts’ao C, Hart SJ, Krajewski DV, Sorensen PG. Opposite effect of lysine on platelet aggregation induced by arachidonate and by other aggregants. Thromb Haemostas 1982; 48: 78-83
  • 23 Livne A, Gartner TK, Agam G. Inhibition of platelet aggregation and activity of the endogenous platelet lectin by oligoamines. Thromb Haemostas 1983; 50: 132 (Abstr)
  • 24 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
  • 25 Kinlough-Rathbone RL, Mustard JF, Packham MA, Perry DW, Reimers H-J, Cazenave J-P. Properties of washed human platelets. Thromb Haemostas 1977; 37: 291-308
  • 26 Kinlough-Rathbone RL, Packham MA, Mustard JF. Synergism between platelet aggregating agents: the role of the arachidonate pathway. Thromb Res 1977; 11: 567-580
  • 27 Vargaftig BB, Chignard M, Benveniste J. Present concepts on the mechanisms of platelet aggregation. Biochem Pharmacol 1981; 30: 263-271
  • 28 Henson PM, Ginsberg MH. Immunological reactions of platelets. Gordon JL. (ed) In: Platelets in biology and pathology. Oxford, Elsevier/North Holland; 1981. 2 265-308
  • 29 Cazenave J-P, 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
  • 30 Pinckard RN, McManus LM, Halonen M, Hanahan DJ. Acetyl glyceryl ether phosphorylcholine: platelet-activating factor. Int Arch Allergy Appl Immunol 1981; 66 Suppl (Suppl. 01) 127-136
  • 31 Wörner P, Brossmer R. Effect on human platelets of ionophores for divalent cations. Naturwissenschaften 1974; 61: 508-509
  • 32 Feinman RD, Detwiler TC. Absence of requirement for extracellular calcium for secretion from platelets. Thromb Res 1975; 7: 677-679
  • 33 Rink TJ, Smith SW, Tsien RY. Intracellular free calcium in platelet shape change and aggregation. J Physiol 1982; 324: 53P-54P
  • 34 Gerrard JM, Peterson DA, White JG. Calcium mobilization. Gordon JL. (ed) In Platelets in biology and pathology. Oxford, Elsevier/North Holland; 1981. 2 407-436