Thromb Haemost 1969; 22(02): 304-315
DOI: 10.1055/s-0038-1651356
Originalarbeiten-Original Articles-Travaux Originaux
Schattauer GmbH

Platelet Incorporation of Labelled Adenosine and Adenosine Diphosphate

E. W Salzman
1   Currently at Columbia University, New York, New York.
,
T. P Ashford
1   Currently at Columbia University, New York, New York.
,
D. A Chambers
1   Currently at Columbia University, New York, New York.
,
Lena L. Neri
1   Currently at Columbia University, New York, New York.
› Author Affiliations
This investigation was supported by Grant No. HE-11127 and Grant No. HE-06316 from the National Heart Institute.
Further Information

Publication History

Publication Date:
10 June 2018 (online)

Summary

After incubation of platelet-rich plasma with labelled adenosine or ADP, platelet incorporation of radioactivity was assessed. Platelets were rapidly separated for counting by filtration through cellulose acetate Millipore. Inulin-H3 served as a plasma marker, and triple isotope techniques permitted simultaneous assessment of the behavior of the adenine and phosphate moieties of ADP without washing of platelets. In other experiments, electron microscopic radioautography was employed to trace the label after platelet incorporation.

The results were consistent with previous reports that ADP is dephosphorylated in plasma and is incorporated by platelets only as a dephosphorylated residue, probably adenosine. The label crossed the platelet membrane and entered the platelet, where it was distributed in platelet granules and the agranular cell sap. Concentration within granules occurred to a minor degree.

The results support the hypothesis that platelet aggregation by ADP occurs without a persistent bond of ADP to the platelet. Inhibition of aggregation by adenosine probably depends on a metabolic or transport process rather than on competition between adenosine and ADP for platelet binding sites.

 
  • References

  • 1 Baker R. V, Blaschko H, Born G. V. R. Isolation from blood platelets containing 5-hydroxy-tryptamine and adenosine triphosphate. J. Physiol. (Lond.) 149: 55 1959;
  • 2 Born G. V. R. Uptake of adenosine and andenosme diphosphate by human blood platelets. Nature (Lond.) 206: 1121 1965;
  • 3 Born G. V. R. Platelet membrane and its functions. XII Congr. Internat. Soc. Hematol, New York, 1968 ; Plenary Session Papers, p. 95.
  • 4 Born G. V. R, Cross M. J. The aggregation of blood platelets. J. Physiol. (Lond.) 168: 178 1963;
  • 5 Born G. V. R, Gillson R. E. Studies on the uptake of 5-hydroxy-tryptamine by blood platelets. J. Physiol. (Lond.) 146: 472 1959;
  • 6 Garo L. G. The problem of resolution in high resolution autoradiography. J. Cell. Biol. 15: 189 1962;
  • 7 Garo L. G, Van Tubergen R. P, Kolb J. A. High resolution autoradiography. I. Methods. J. Cell Biol. 15: 173 1962;
  • 8 Chambers D. A, Salzman E. W, Neri L. L, Sivertsen U. Platelet incorporation of fibrinogen I125 . XII Congr. Internat. Soc. Hematol., New York, 1968 ; Plenary Session Papers, p. 200.
  • 9 Gaarder A, Laland S. Hypothesis for aggregation of platelets by nucleotides. Nature (Lond.) 202: 909 1964;
  • 10 Grette K. Studies on the mechanism of thrombin-catalyzed hemostatic reactions in blood platelets. Acta physiol. scand. 56 (Suppl. 195) 1 1962;
  • 11 Hampton J. R, Mitchell J. R. A. Effect of aggregating agents on the electrophoretic mobility of human platelets. Brit. med. J. I: 1074 1966;
  • 12 Haslam R. J. Biochemical aspects of platelet function. XII Congr. Internat. Soc. Hematol., New York, 1968 ; Plenary Session Papers, p. 198.
  • 13 Holmsen H. Adenine nucleotide pools in platelets. XII Congr. Internat. Soc. Hematol., New York, 1968 ; Plenary Session Papers, p. 202.
  • 14 Hovig T. Release of a platelet-aggregating substance (adenosine diphosphate) from rabbit blood platelets induced by saline ‘extract’ of tendons. Thrombos. Diathes. haemorrh. (Stuttg.) 8: 264 1963;
  • 15 Ireland D. M. Effect of thrombin on radioactive nucleotides of human washed platelets. Biochem. J. 105: 857 1967;
  • 16 Luft J. H. Improvements in epoxy resin embedding methods. J. biophys. biochem. Cytol. 9: 409 1961;
  • 17 Markwardt F, Barthel W, Glusa E, Hoffman A. Untersuchungen über den Einfluß von Papaverin auf Reaktionen der Blutplättchen. Naunyn-Schmiedebergs Arch. Pharmak. exp. Path. 257: 420 1967;
  • 18 Millonig G. A modified procedure for lead staining of thin sections. J. biophys. biochem. Cytol. 11: 736 1961;
  • 19 Moses M. J. Application of autoradiography to electron microscopy. J. Histochem. Cyto-chem. 12: 115 1964;
  • 20 O’Brien J. R, Heywood J. B. Effect of aggregating agents and their inhibitors on the mean platelet shape. J. clin. Path. 19: 148 1966;
  • 21 Richardson K. C, Jarett L, Finke E. H. Embedding in epoxy resins for ultra-thin sectioning in electron microscopy. Stain Technol. 35: 313 1960;
  • 22 Rozenberg M. C, Holmsen H. Adenine nucleotide metabolism of blood platelets. II. Uptake of adenosine and inhibition of ADP-induced platelet aggregation. Biochim. biophys. Acta (Amst.) 155: 342 1968;
  • 23 Sabatini D. D, Bensch K. G, Barnett R. J. New means of fixation for electron microscopy and histochemistry. Anat. Ree. 142: 274 1962;
  • 24 Salzman E. W, Chambers D. A, Neri L. L. Incorporation of labelled nucleotides and aggregation of human blood platelets. Thrombos. Diathes. haemorrh. (Stuttg.) 15: 52 1966;
  • 25 Salzman E. W, Chambers D. A, Neri L. L. Possible mechanisms of aggregation of blood platelets by adenosine diphosphate. Nature (Lond.) 210: 167 1966;
  • 26 Salzman E. W, Neri L. L, Chambers D. A, Ashford T. P. Platelet volume: alteration by changes in temperature and agents affecting platelet aggregation. Amer. J. Physiol. (In press.)
  • 27 Skoza L, Zucker J, Jerushalmy Z, Grant R. Kinetic studies of platelet aggregation induced by adenosine diphosphate and its inhibition by chelating agents, guanidino compounds, and adenosine. Thrombos. Diathes. haemorrh. (Stuttg.) 18: 713 1967;
  • 28 Spaet T. H, Lejnieks I. Studies on the mechanism whereby platelets are clumped by adenosine diphosphate. Thrombos. Diathes. haemorrh. (Stuttg.) 15: 36 1966;