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Thromb Haemost 1999; 82(04): 1318-1321
DOI: 10.1055/s-0037-1614383
Review Article
Schattauer GmbH

Platelet Aging In Vivo Is Associated with Loss of Membrane Phospholipid Asymmetry

Jaime Pereira
1   From the Department of Hematology-Oncology, School of Medicine, Catholic University of Chile, Santiago
,
Iván Palomo
2   Department of Clinical Sciences, Faculty of Health Sciences, University of Talca, Chile
,
Mauricio Ocqueteau
1   From the Department of Hematology-Oncology, School of Medicine, Catholic University of Chile, Santiago
,
Mónica Soto
1   From the Department of Hematology-Oncology, School of Medicine, Catholic University of Chile, Santiago
,
Eduardo Aranda
1   From the Department of Hematology-Oncology, School of Medicine, Catholic University of Chile, Santiago
,
Diego Mezzano
1   From the Department of Hematology-Oncology, School of Medicine, Catholic University of Chile, Santiago
› Author Affiliations
Further Information

Publication History

Received 08 March 1999

Accepted after revision 06 May 1999

Publication Date:
08 December 2017 (online)

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Summary

The mechanism(s) involved in the clearance of senescent platelets are largely unknown. The loss of membrane phospholipid (PL) asymmetry, with phosphatidylserine (PS) exposure appears to be an important signal for the ingestion by macrophages of apoptotic nucleated cells and it has also been suggested as a signal for the removal of aged erythrocytes. Accordingly, it seems possible that the clearance of normal aged platelets from circulation might be triggered by PS exposure. To investigate this, we determined PS exposure in human aging platelets taking advantage of the relationship between platelet density and platelet age and in dog platelets in a model of platelet aging in vivo. PS exposure was determined in two experimental conditions: 1) human platelet density subpopulations obtained by centrifugation in arabinogalactan gradients; 2) circulating canine platelets during decline in platelet count after suppression of thrombopoiesis following estradiol injection. PS exposure was determined by flow cytometry after labeling the cells with FITC-conjugated annexin V. The proportion of human platelets with exposed PS was significantly higher in high density (HD) platelets compared to low density (LD) platelets (11.3 ± 8.0% vs 5.2 ± 3.7%; p <0.05, respectively). In dogs, the proportion of cells with exposed PS rises dramatically with age, from 3.1 ± 0.4% before to 17.7 ± 12.3% ten days after estradiol injection. These findings suggest that platelet aging is associated with loss of phospholipid asymmetry and PS exposure on the outer leaflet of cell membrane, which may play an important role in the recognition and subsequent removal of senescent platelets.

Key Words

Platelet age - phosphatidylserine - platelet removal
 

  • References

  • 1 George JN, Dale GL. Platelet kinetics. In: Hematology. Beutler E, Lichtman MA, Coller BS, Kipps TJ. eds. New York: McGraw-Hill Inc; 1995: 1202-5.
    Search in Google Scholar
  • 2 Mezzano D, Hwang KL, Catalano P, Aster RH. Evidence that platelet buoyant density, but not size, correlates with platelet age in man. Am J Hematol 1981; 11: 61-76.
    CrossrefPubMedSearch in Google Scholar
  • 3 Boneu B, Vigoni F, Boneu A, Caranobe C, Sie P. Further studies on the relationship between platelet buoyant density and platelet age. Am J Hematol 1982; 13: 239-46.
    CrossrefPubMedSearch in Google Scholar
  • 4 Mezzano D, Aranda E, Rodríguez S, Foradori A, Lira P. Increase in density and accumulation of serotonin by human aging platelets. Am J Hematol 1984; 17: 11-21.
    CrossrefPubMedSearch in Google Scholar
  • 5 Martin JF, Penington DG. The relationship between the age and density of circulating 51 Cr-labeled platelets in the subhuman primate. Thromb Res 1983; 30: 157-64.
    CrossrefPubMedSearch in Google Scholar
  • 6 Karpatkin S. Heterogeneity of rabbit platelets VI. Further resolution of changes in platelet density, volume, and radioactivity following cohort labeling with 75 Se-seleniomethionine. Br J Haematol 1978; 39: 459-69.
    CrossrefPubMedSearch in Google Scholar
  • 7 Corash L, Costa JL, Shafer B, Donlon JA, Murphy D. Heterogeneity of human whole blood platelet subpopulations. III. Density-dependent differences in subcellular constituents. Blood 1984; 64: 185-193.
    PubMedSearch in Google Scholar
  • 8 Packham MA, Guccione MA, O’Brien KM. Duration of the effect of aspirin on the synthesis of thromboxane by density subpopulations of rabbit platelets stimulated by thrombin. Blood 1985; 66: 287-90.
    PubMedSearch in Google Scholar
  • 9 Mezzano D, Aranda E, Foradori A, Rodríguez S, Lira P. Kinetics of platelet density subpopulations in splenectomized mongrel dogs. Am J Hematol 1984; 17: 373-82.
    CrossrefPubMedSearch in Google Scholar
  • 10 Mishory B, Danon D. Structural aspects of in vivo aging rabbit blood platelets. Thromb Res 1978; 12: 893-906.
    CrossrefPubMedSearch in Google Scholar
  • 11 Mezzano D, Aranda E, Foradori A. Comparative study of size, total protein, fibrinogen and 5-HT content of human and canine platelet density subpopulations. Thromb Haemost 1986; 56: 288-92.
    Thieme ConnectPubMedSearch in Google Scholar
  • 12 Kelton JG, Denomme G. The quantitation of platelet-associated IgG on cohorts of platelets separated from healthy individuals by buoyant density centrifugation. Blood 1982; 60: 136-9.
    PubMedSearch in Google Scholar
  • 13 Pfueller SL, Chesterman C, Illes I, Hussein S, Martin JF. Relationship of platelet-associated immunoglobulin G and platelet protein to platelet size and density in normal individuals and patients with thrombocytopenia. J Lab Clin Med 1986; 107: 299-305.
    PubMedSearch in Google Scholar
  • 14 Pereira J, Cretney C, Aster RH. Variation of class I HLA antigen expression among platelet density cohorts: A possible index of platelet age?. Blood 1988; 71: 516-9.
    PubMedSearch in Google Scholar
  • 15 Aranda E, Pereira J, Ajenjo C, Prieto C, Sepúlveda S, Mezzano D. Human intraplatelet 5-hydroxytryptamine is correlated with mean platelet survival time. Thromb Res 1996; 84: 67-72.
    CrossrefPubMedSearch in Google Scholar
  • 16 Aranda E, Pizarro M, Pereira J, Mezzano D. Accumulation of 5-hydroxy-tryptamine by aging platelets: studies in a model of supressed thrombopoiesis in dogs. Thromb Haemost 1994; 71: 488-92.
    Thieme ConnectPubMedSearch in Google Scholar
  • 17 Martin SJ, Reutelingsperger CPM, McGahon AJ, Rader JA, VanSchie RCAA, Laface DM, Green DR. Early redistribution of plasma membrane phosphatidylserine is a general feature of apoptosis regardless of the initiating stimulus: Inhibition by overexpression of Bcl-2 and abl. J Exp Med 1995; 182: 1545-56.
    CrossrefPubMedSearch in Google Scholar
  • 18 Zwaal RFA, Schroit AJ. Pathophysiologic implications of membrane phospholipid asymmetry in blood cells. Blood 1997; 89: 1121-32.
    PubMedSearch in Google Scholar
  • 19 Bevers EM, Comfurius P, Zwaal RFA. Changes in membrane phospholipid distribution during platelet activation. Biochim Biophys Acta 1983; 736: 57-66.
    CrossrefPubMedSearch in Google Scholar
  • 20 Blumenfeld N, Zachowski A, Galacteros F, Beuzard Y, Devaux PF. Transmembrane mobility of phospholipids in sickle erythrocytes: effect of de-oxygenation on diffusion and asymmetry. Blood 1991; 77: 849-54.
    PubMedSearch in Google Scholar
  • 21 Connor J, Pak CC, Schroit AJ. Exposure of phosphatidylserine in the outer leaflet of human red blood cells: Relationship to cell density, cell age, and clearance by mononuclear cells. J Biol Chem 1994; 269: 2399-404.
    PubMedSearch in Google Scholar
  • 22 Utsugi T, Schroit AJ, Connor J, Bucana CD, Fidler IJ. Elevated expression of phosphatidylserine in the outer membrane leaflet of human tumor cells and recognition by activated human monocytes. Cancer Res 1991; 51: 3062-6.
    PubMedSearch in Google Scholar
  • 23 Fadok VA, Savill JS, Haslett C, Bratton DL, Doherty DE, Campbell PA, Henson PM. Different populations of macrophages use either the vitronectin receptor or the phosphatidylserine receptor to recognize and remove apoptotic cells. J Immunol 1992; 149: 4029-35.
    PubMedSearch in Google Scholar
  • 24 Savill J, Fadok V, Henson P, Haslett C. Phagocyte recognition of cells undergoing apoptosis. Immunol Today 1993; 14: 131-6.
    CrossrefPubMedSearch in Google Scholar
  • 25 Dachary-Prigent J, Freyssinet JM, Pasquet JM, Carron JC, Nurden AT. Annexin V as a probe of aminophospholipid exposure and platelet membrane vesiculation: a flow cytometric study showing a role for free sulfhydryl groups. Blood 1993; 81: 2554-65.
    PubMedSearch in Google Scholar
  • 26 Vanags DM, Orrenius S, Aguilar-Santelices M. Alterations in Bcl-2/Bax protein levels in platelets form part of an ionomycin-induced process that resembles apoptosis. Br J Haematol 1997; 99: 824-31.
    CrossrefPubMedSearch in Google Scholar
  • 27 Chiu T. Studies on estrogen-induced proliferative disorders of hemopoietic tissue in dogs. St. Paul: University of Minnesota. Ph. D. Thesis 1974: 1-270.
    PubMedSearch in Google Scholar
  • 28 Kumar A, Kumar A, Deepika K, Fernández J, Eisdorfer C. A modified HPLC technique for simultaneous measurement of 5-hydroxytryptamine and 5-hydroxyindolacetic acid in cerebrospinal fluid, platelet and plasma. Life Sciences 1990; 47: 1751-9.
    CrossrefPubMedSearch in Google Scholar
  • 29 Hale AJ, Smith CA, Sutherland LC, Stoneman VE, Longthorne VL, Culhane AC, Williams GT. Apoptosis: molecular regulation of cell death. Eur J Biochem 1996; 236: 1-26.
    CrossrefPubMedSearch in Google Scholar
  • 30 Tonon G, Luo X, Chen W, Shi Y, Greco N, Cattaneo M, Jamieson GA. Expression of apoptotic markers in activated platelets is mediated in part by cathepsin. Blood 1998; 92: 30a.
    PubMedSearch in Google Scholar
  • 31 Berger G, Hartwell DW, Wagner DD. Platelet death. An apoptotic like process?. Blood 1998; 92: 347a.
    PubMedSearch in Google Scholar
  • 32 Susin SA, Zamzami N, Kroemer G. Mitochondria as regulators of apoptosis: doubt no more. Biochim Biophys Acta 1998; 1366: 151-65.
    CrossrefPubMedSearch in Google Scholar