Assessment of Damage to Human Platelets after Aggregation and Other Injuries by Microscopic Observation and Estimation of Serotonin Uptake

G Zbinden; J. N Mehrishi; S Tomlin

doi:10.1055/s-0038-1654139

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1970; 23(02): 261-275
DOI: 10.1055/s-0038-1654139

Originalarbeiten – Original Articles – Travaux Originaux

Schattauer GmbH

Assessment of Damage to Human Platelets after Aggregation and Other Injuries by Microscopic Observation and Estimation of Serotonin Uptake

Authors

G Zbinden¹
J. N Mehrishi

^*Departments of Medicine and Radiotherapeutics*, University of Cambridge, Cambridge
S Tomlin

Abstract

Summary

The severity of platelet damage induced by hyper- and hypotonic NaCl solutions and freezing and thawing was assessed by microscopic evaluation and measuring inhibition of 5-HT uptake. The same techniques were used to quantitate the effects of aggregating agents. The positively charged macromolecules PS, Poly-L und Poly-O reduced the net negative surface charge as determined by microelectrophoresis, caused platelet aggregation and inhibited 5-HT uptake. The damaging effects of Poly-L and Poly-O were more severe and more closely related to concentration than that of PS. The negatively charged macromolecules Poly-IC and NaPS increased the anodic electrophoretic mobility. Poly-IC and heparin caused a low degree of platelet clumping and no inhibition of 5-HT uptake. NaPS produced severe platelet damage with extensive clumping and complete inhibition of 5-HT uptake. Na laurate had the same effect, but did not alter electrophoretic mobility. ADP caused concentration-dependent platelet aggregation and inhibition of 5-HT uptake. The effects of ADP and NaPS were compared in agitated and non-agitated platelet samples containing identical concentrations of the 2 compounds. Agitation was found to increase the degree of platelet clumping and to reduce 5-HT uptake.

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Referenzen

References
1 Bangham A. D, Pethica B. A, Seaman G. V. F. The charged groups at the interface of some blood cells. Biochem. J 69: 12-19 1958;
2 Baumgartner H. R, Tranzer J. P, Studer A. An electron microscopic study of platelet thrombus formation in the rabbit with particular regard to 5-hydroxytryptamine release. Thrombos. Diathes. haemorrh. (Stuttg.) 13: 592-604 1967;
3 Bettex-Galland M, Lüscher E. F, Simon G, Vasalli P. Induction of viscous metamorphosis in human blood platelets by means other than thrombin. Nature (Lond.) 200: 1109-1110 1963;
4 Betts J. J, Betts J. P, Nicholson J. T. Significance of ADP, plasma and platelet concentration in platelet electrophoretic studies. Nature (Lond.) 219: 1280-1282 1968;
5 Born G. V. R. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature (Lond.) 194: 927-929 1962;
6 Born G. V. R. Quantitative investigations into the aggregation of blood platelets. J. Physiol 162: 67P-68P 1962;
7 Born G. V. R. The platelet membrane and its function. In: Metabolism and Membrane Permeability of Erythrocytes and Thrombocytes. 1st International Symposium, Vienna June 17-20, 1968. Deutsch E, Gerlach D, Moser K. eds. 294-301 Georg Thieme; Stuttgart:
8 Born G. V. R, Gillson R. E. Studies on the uptake of 5-hydroxytryptamine by blood platelets. J. Physiol 146: 472-491 1959;
9 Boidlin D. J, O’Brien R. A. The binding of Imipramine to the outer membrane of blood platelets. J. Pharm. Pharmacol 20: 583-584 1968;
10 Breddin K. Reversible und irreversible Funktionsstörungen der Thrombocyten. In: Metabolism and Membrane Permeability of Erythrocytes and Thrombocytes. 1st International Symposium, Vienna June 17-20, 1968. Deutsch E, Gerlach E, Moser K. eds. 307-313 Georg Thieme; Stuttgart:
11 Bull S. B, Zucker M. B. C. Changes in platelet volume produced by temperature, metabolic inhibitors and aggregating agents. Proc. Soc. exp. Biol. (N. Y.) 120: 296-301 1965;
12 DaPrada M, Tranzer J. P, Pletscher A. Effect of drugs and cations on ultrastructure and 5-hydroxytryptamine content of blood platelets. J. Pharmacol, exp. Ther 158: 394-398 1967;
13 DaPrada M, Pletscher A. Isolated 5-hydroxytryptamine organelles of rabbit blood platelets : physiological properties and drug-induced changes. Brit. J. Pharmacol 34: 591-597 1968;
14 Gaarder A, Laland S. Hypothesis for the aggregation of platelets by nucleotides. Nature (Lond.) 202: 909-910 1964;
15 Grette K. Studies on the mechanism of thrombin-catalyzed reactions in blood platelets. Acta physiol. scand 56 (Suppl. 194) 1-93 1962;
16 Gröttum K. A. Platelet surface charge and aggregation. Effects of polyelectrolytes. Thrombos. Diathes. haemorrh. (Stuttg.) 21: 450-462 1969;
17 Hampton J. R, Mitchell J. R. A. Effect of aggregating agents on the electrophoretic mobility of human platelets. Brit. med. J I: 1074-1077 1966;
18 Hampton J. R, Mitchell J. R. A. Modification of the electrokinetic response of blood platelets to aggregating agents. Nature (Lond.) 210: 1000-1002 1966;
19 Haslam R. J. Role of adenosine diphosphate in the aggregation of human blood platelets by thrombin and by fatty acids. Nature (Lond.) 202: 765-768 1964;
20 Hellem A, Owren P. A. The mechanism of the hemostatic function of blood platelets. Acta haemat. (Basel) 31: 230-238 1964;
21 Hovig T. The ultrastructure of rabbit blood platelet aggregates. Thrombos. Diathes. haemorrh. (Stuttg.) 8: 455-471 1962;
22 Kirschmann G, Katchalsky A, de Vries A. Electrochemical study of the platelet surface. Arch. Biochem 80: 140-149 1959;
23 Lohr G. W. Der Stoffwechsel der normalen und pathologischen Thrombocyten. In: Metabolism and Membrane Permeability of Erythrocytes and Thrombocytes. 1st International Symposium, Vienna June 17-20, 1968. Deutsch E, Gerlach E, Moser K. eds. 196-205 Georg Thieme; Stuttgart:
24 Lüscher E. F. Viscous metamorphosis of blood platelets and clot retraction. Vox Sang 1: 133-154 1956;
25 Madoff M. A, Ebbe S, Baldini M. Sialic acid of human blood platelets. J. clin. Invest 43: 870-877 1964;
26 Mehrishi J. N. Positively charged amino groups on the surface of normal and cancer cells. Europ. J. Cancer 6: 127-137 1970;
27a Mehrishi J. N, Grassetti D. R. Sulphydryl groups on the surface of intact Ehrlich ascites tumour cells, human blood platelets and lymphocytes. Nature (Lond.) 224: 563-564 1969;
27b Mehrishi J. N. Phosphate groups (receptors) on the surface of human blood platelets. Nature (Lond.) 226: 452-453 1970;
28 Mehrishi J. N. Effect of lysine polypeptides on the surface charge of normal and cancer cells. Europ. J. Cancer 5: 427-435 1969;
29 Michelson A. M, Pochon F. Polynucleotide analogues. VII. Methylation of polynucleotides. Biochim. biophys. Acta (Amst.) 114: 469-480 1966;
30 Mills D. G. B, Robb I. A, Roberts G. G. K. The release of nucleotides, 5-hydroxytryptamine and enzymes from human blood platelets during aggregation. J. Physiol 195: 715-731 1968;
31 Ozge A, Baldini M, Goldstein R. Effect of plastic and glass surfaces on clot retraction and serotonin uptake of platelet-rich plasma stored at 4° C. J. Lab. clin. Med 63: 378-393 1964;
32 Reuter H, Niemeyer G, Gross R. Zut Differenzierung der Plattchenaggregation. In: Metabolism and Membrane Permeability of Erythrocytes and Thrombocytes. 1st International Symposium, Vienna June 17-20, 1968. Detitsch E, Gerlaeh E, Moser K. eds. 335-338 Georg Thieme; Stuttgart:
33 Salzman E. W. Studies on the mechanism of platelet aggregation by adenosine diphosphate. In: Physiology of Hemostasis and Thrombosis. Johnson S. A, Seegers W. H. eds. 140-153 Charles C. Thomas Pubi.; Springfield, Ill.:
34 Salzman E. W, Chambers D. A, Neri L. L. Possible mechanism of aggregation of blood platelets by adenosine diphosphate. Nature (Lond.) 210: 167-169 1966;
35 Salzman E. W, Neri L. L, Chambers D. A, Ashford T. P. Regulation of platelet volume. In: Metabolism and Membrane Permeability of Erythrocytes and Thrombocytes. 1st International Symposium Vienna June 17-20, 1968. Deutsch E, Gerlaeh E, Moser K. eds. 274-282 Georg Thieme; Stuttgart:
36 Seaman G. V. F. Surface potential and platelet aggregation. Thrombos. Diathes. haemorrh. (Stuttg.) Suppl. 26: 53-68 1967;
37 Seaman G. V. F, Vassar P. S. Changes in the electrokinetic properties of platelets during their aggregation. Arch. Biochem 117: 10-17 1966;
38 Sharp A. A. Viscous metamorphosis of blood platelets: a study of the relationship to coagulation factors and fibrin formation. Brit. J. Haemat 4: 28-37 1958;
39 Shore P. A, Alpers H. S. Parallel inhibition of spontaneous and antigen-induced platelet histamine release. Amer. J. Physiol 205: 348-350 1963;
40 Shore P. A, Alpers H. S. Platelet damage and release of amines by microgram quantities of certain fatty acids. Fed. Proc 22: 504 1963;
41 Solomon H. M, Ashley C, Barkins G, Spirt N. The effect of debrisoquin on the human platelet. Pharmacologist 10: 211 1968;
42 Solum N. O, Stormorken H. Influence of fibrinogen on the aggregation of washed human blood platelets induced by adenosine diphosphate, thrombin and adrenaline. Scand. J. clin. Lab. Invest 17: 170-182 1965;
43 Spaet T. H, Lejnicks I. Studies on the mechanism whereby platelets are clumped by adenosine diphosphate. Thrombos. Diathes. haemorrh. (Stuttg.) 15: 36-51 1966;
44 Stacey R. S. Uptake of 5-hydroxytryptamine by platelets. Brit. J. Pharmacol 16: 284-295 1961;
45 Szent-Györgyi A. Bioelectronics. A study in cellular regulations, defense and cancer. Academic Press; London: 1968
46 Telkää A, Nyholm M, Paasonen M. K. Effect of chlorpromazine and reserpine on the blood platelet of rabbit. An electron microscopic study. Experientia (Basel) 20: 27-28 1964;
47 Tranzer J. P, Pletscher A, DaPrada M. Speicherung von 5-Hydroxytryptamin in submikroskopischen Organellen der Blutplättchen. Helv. physiol. Acta 24: C108-C110 1966;
48 Tranzer J. P, DaPrada M, Pletscher A. Ultrastructural localisation of platelet 5-HT. Nature (Lond.) 212: 1574-1575 1966;
49 Weissbach H, Redfield B. G. Factors affecting the uptake of 5-hydroxytryptamine by human platelets in an inorganic medium. J. biol. Chem 235: 3287-3291 1960;
50 White J. G. Fine structural alterations induced in platelets by adenosine diphosphate. Blood 31: 604-622 1968;
51 Zbinden G, Tomlin S. Effect of debrisoquin, guanethidine and tyramine on platelet function and blood coagulation. Arch. int. Pharmacodyn. (In press.)
52 Zucker M. B, Borelli J. Viscous metamorphosis, clot retraction and other morphologic alterations of blood platelets. J. appl. Physiol 14: 575-580 1959;