Morphometric Analysis of Density Subpopulations of Normal Human Platelets

Summary

Platelets from seven normal subjects were fractionated on continuous Percoll density gradients and low density (LD), intermediate, and high density (HD) platelets were prepared for transmission electron microscopy followed by computerised morphometric analysis. Normal ultrastructural appearance and discoid shape were preserved by incubation of the platelets in nutrient medium at 37 °C immediately before fixation. HD platelet sections had a larger mean cross-sectional area but a lower ratio of the major to the minor axis compared to LD platelet sections. HD platelets contained more alpha granules, dense granules and mitochondria per square micron of section area than LD platelets. The percentage of section area occupied by open canalicular system was greater in the LD platelets while the percentage area occupied by glycogen fields was over ten-fold higher in the HD platelets. The mean cross-sectional areas of individual alpha granules and dense granules increased with density while the opposite trend was found for mitochondria. It is suggested that these ultrastructural differences mainly arise during thrombopoiesis and may indicate some functional specialization among platelets.

Key words

Platelet density - Platelet ultrastructure - Morphometric analysis

• References

• 1 Martin JF, Shaw T, Heggie J, Penington DG. Measurement of the density of human platelets and its relationship to volume. Br J Haematol 1983; 54: 337-352
• 2 Van Oost B, Timmermans AP M, Sixma JJ. Evidence that platelet density depends on the α-granule content in platelets. Blood 1984; 63: 482-485
• 3 Bolin RB, Cheney BA, Simpliciano OA, Smith DJ. Buoyant density of platelets stored at room temperature as platelet concentrates. J Lab Clin Med 1981; 98: 342-351
• 4 Rendu F, Nurden AT, Lebret M, Caen JP. Relationship between mepacrine-labelled dense body number, platelet capacity to accumulate ¹⁴C-5-HT and platelet density in the Bemard-Soulier and Hermansky-Pudlak syndromes. Thromb Haemostas 1979; 42: 694-704
• 5 Vicie WJ, Weiss HJ. Evidence that platelet α-granules are a major determinant of platelet density: studies in storage pool deficiency. Thromb Haemostas 1983; 50: 878-880
• 6 Caranobe C, Sie P, Nouvel C, Laurent G, Pris J, Boneu B. Platelets in myeloproliferative disorders. II. Serotonin uptake and storage: correlations with mepacrine labelled dense bodies and with platelet density. Scand J Haematol 1980; 25: 289-295
• 7 Van Oost B, van Hien-Hagg IR, Timmermans AP M, Sixma JJ. The effect of thrombin on the density distribution of blood platelets: detection of activated platelets in the circulation. Blood 1983; 62: 433-438
• 8 Boneu B, Sie P, Eche N, Caranobe C, Hugo B, Nouvel C. Platelet density analysis: a tool for the detection of acquired storage pool disease in man. Br J Haematol 1983; 55: 523-532
• 9 Charmatz A, Karpatkin S. Heterogeneity of rabbit platelets. I. Employment of an albumin density gradient for separation of a young platelet population identified with ⁷⁵Se-selenomethionine. Thromb Diath Haemorrh 1974; 31: 485-492
• 10 Rand ML, Greenberg JP, Packham MA, Mustard JF. Density subpopulations of rabbit platelets: size, protein, and sialic acid content, and specific radioactivity changes following labelling with ³⁵S-sulphate in vivo. Blood 1981; 57: 741-746
• 11 Corash L, Shafer B. Use of asplenic rabbits to demonstrate that platelet age and density are related. Blood 1982; 60: 166-171
• 12 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
• 13 Mezzano D, Aranda E, Rodriguez S, Foradori A, Lira P. Increase in density and accumulation of serotonin by human ageing platelets. Am J Hematol 1984; 17: 11-21
• 14 Boneu B, Sie P, Caranobe C, Nouvel C, Bierme R. Malondialdehyde (MDA) reappearance in human platelet density subpopulations after a single intake of aspirin. Thromb Res 1980; 19: 609-620
• 15 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: 239-246
• 16 McDonald JW D, Ali M. Recovery of cyclo-oxygenase activity after aspirin in populations of platelets separated on Stractan density gradients. Prostaglandins, Leukotrienes Med 1983; 12: 245-252
• 17 Chamberlain KG, Chiu E, Penington DG. Relationship of platelet density to ageing. Cohort studies using monoamine oxidase activity Pathology 1984; 16: 355 (abstract)
• 18 Penington DG, Streatfield K, Roxburgh AE. Megakaryocytes and the heterogeneity of circulating platelets. Br J Haematol 1976; 34: 639-653
• 19 Corash L, Tan H, Gralnick HR. Heterogeneity of human whole blood platelet subpopulations. I. Relationship between buoyant density, cell volume, and ultrastructure. Blood 1977; 49: 71-87
• 20 Haver VM, Gear AR L. Functional fractionation of platelets. J Lab Clin Med 1981; 97: 187-204
• 21 Chamberlain KG, Penington DG. Monoamine oxidase and other mitochondrial enzymes in density subpopulations of human platelets. Thromb Haemostas 1988; 59: 29-33
• 22 Mayhew TM. A comparison of several methods for stereological determination of the numbers of organelles per unit volume of cytoplasm. J Microsc 1972; 96: 37-44
• 23 Zucker WH, Shermer RW, Mason RG. Ultrastructural comparison of human platelets separated from blood by various means. Am J Pathol 1974; 77: 255-268
• 24 Mustard JF, Perry DW, Ardlie NC, Packham MA. Preparation of suspensions of washed platelets from humans. Br J Haematol 1972; 22: 193-204
• 25 White JG. Fine structural alterations induced in platelets by adenosine diphosphate. Blood 1968; 31: 604-622
• 26 Corash L, Costa JL, Shafer B, Danlon JA, Murphy D. Heterogeneity of human blood platelet subpopulations. III. Density-dependent differences in subcellular constituents. Blood 1984; 64: 185-193
• 27 Lewis JC, Maldonado JE, Mann KG, Moertel CG. Ultrastructural cytochemistry of platelets and megakaryocytes in the carcinoid syndrome. Mayo Clin Proc 1976; 51: 585-593
• 28 Gerrard JM, Rao GH R, White JG. The influence of reserpine and ethylenediaminetetraacetic acid on serotonin storage organelles of blood platelets. Am J Pathol 1977; 87: 633-646
• 29 Mishory B, Danon D. Structural aspects of in vivo rabbit blood platelets. Thromb Res 1978; 12: 893-906
• 30 Lorez HP, Da Prada M, Rendu F, Pletscher A. Mepacrine, a tool for investigating the 5-hydroxytryptamine organelles of blood platelets by fluorescence microscopy. J Lab Clin Med 1977; 89: 200-206
• 31 Caranobe C, Sie P, Boneu B. Serotonin uptake and storage in human platelet density subpopulations. Br J Haematol 1982; 52: 253-258
• 32 Stahl K, Themann H, Dame WR. Ultrastructural morphometric investigations on normal human platelets. Haemostasis 1978; 7: 242-251
• 33 Kainuma S, Deguchi K, Kobayashi T, Murashima M, Nagano T, Shirakawa S. Morphometric analysis of platelet organelles in various hemorrhagic diathesis. J Clin Electron Microsc 1981; 14: 704-705
• 34 Payne CM, Glasser L. An ultrastructural morphometric analysis of platelet giant and fusion granules. Blood 1986; 67: 299-309
• 35 Karpatkin S, Charmatz A. Heterogeneity of human platelets. III. Glycogen metabolism in platelets of different sizes. Br J Haematol 1970; 19: 135-143
• 36 Jenkins WJ, Clarkson E, Milson J. et al Simple analytical subcellular fractionation of liver biopsies with Percoll. Clin Sci 1979; 57: 29 P (abstract)
• 37 Gogstad GO. A method for the isolation of α-granules from human platelets. Thromb Res 1980; 20: 669-681
• 38 Scott RB, Still WJ S. Glycogen in human blood platelets. Isolation by ultracentrifugation and characteristics of the isolated particles. Blood 1970; 35: 517-532
• 39 Gear AR L. Preaggregation reactions of platelets. Blood 1981; 58: 477-490
• 40 Karpatkin S. Studies on human platelet glycolysis. Effect of glucose, cyanide, insulin, citrate, and agglutination and concentration on platelet glycolysis. J Clin Invest 1967; 46: 409-417