Gold-Labeled Bovine Fibrinogen for Study of Human Platelets

Summary

Fibrinogen coupled to colloidal gold has proven to be a useful agent for probing the glycoprotein IIb-IIIa receptor on human and bovine platelets at the ultrastructural level. The reagent has helped to demonstrate fundamental differences in the reorganization of the fibrinogen receptors on human and cattle platelets following surface activation. However, commercial preparations of human fibrinogen have not yielded a stable reagent in our hands when coupled to colloidal gold. The present study has substituted bovine for human fibrinogen. Bovine fibrinogen gold proved to be a more stable reagent and could be substituted for human fibrinogen gold in all experiments on human and bovine platelets.

Keywords

GPIIb-IIIa - Fibrinogen binding - Electron microscopy - Gold labeling

• References

• 1 Loftus JC, Albrecht RV. Redistribution of the fibrinogen receptor of human platelets after surface activation. J Cell Biol 1984; 99: 822-829
• 2 Goodman SL, Albrecht RM. Correlative light and electron microscopy of platelet adhesion and fibrinogen receptor expression using colloidal-gold labeling. Scanning Microscopy 1987; 1: 727-734
• 3 Oliver JA, Albrecht RM. Colloidal gold labeling of fibrinogen receptors in epinephrine and ADP-activated platelet suspensions. Scanning Microscopy 1987; 1: 745-756
• 4 Leistikow EA, Barnhart MI, Albrecht MR, White JG. Redistribution of fibrinogen receptors on surface activated platelets. In: Fibrinogen 3. Biochemistry, Biological Functions, Gene Regulation and Expression. Mossesson MW. (ed.) Elsevier Science Publishers; New York: 1988: 215-220
• 5 Escolar G, Leistikow E, White JG. Fate of the platelet open canalicular system (OCS) during surface activation. Circulation 1988; 78: 11546
• 6 Escolar G, Leistikow EA, White JG. Human platelet fibrinogen receptors do not undergo redistribution during surface activation. Blood 1988; 72: 321 a
• 7 White JG, Escolar G, Leistikow EA. GPIIb-IIIa receptor mobility within membranes of surface and suspension activated platelets does not depend on assembly and contraction of cytoplasmic actin. Clin Res 1989; 37: 603 A
• 8 Grouse LH, Rao GH R, Weiss DJ, Perman V, White JG. Surface activated bovine platelets do not spread, they unfold. Am J Pathol (in press)
• 9 White JG. Fine structural alterations induced in platelets by adenosine diphosphate. Blood 1968; 31: 604-622
• 10 White JG, Sauk JJ. Microtubule coils in spread blood platelets. Blood 1984; 64: 470-478
• 11 Gerrard JM, Phillips DR, Rao GH R, Plow EF, Walz DA, Ross R, Harker LA, White JG. Biochemical studies of two patients with the gray platelet syndrome - selective deficiency of platelet alpha granules. J Clin Invest 1980; 109: 102-109
• 12 Rao GH R, Reddy KR, White JG. Modification of human platelet response to sodium arachidonate by membrane modulation. Prost Med 1981; 6: 75-90
• 13 Horisberger MJ, Rossel J, Bauer H. Colloidal gold granules as markers for cell surface receptors in the scanning electron microscope. Experientia 1975; 31: 1147-1151
• 14 Geoghegan W, Ackerman GA. Absorption of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of ConA, wheat germ agglutinin and goat antihuman immunoglobulin G and cell surfaces at the electron microscopic level: A new method, theory and application. J Histochem Cytochem 1977; 25: 1187-1192
• 15 White JG. An overview of platelet structural physiology. Scanning Microscopy 1987; 1: 1677-1700
• 16 Zucker MB, Borelli J. Reversible alterations in platelet morphology produced by anticoagulants and by cold. Blood 1954; 30: 625-630
• 17 Zucker MB, Nachmias VT. Platelet activation. Arteriosclerosis 1985; 5: 2-18
• 18 Nachmias VT. Platelet and megakaryocyte shape change: Triggered alterations in the cytoskeleton. Sem Hematol 1983; 20: 261-281