Localization of fibrinogen during aggregation of avian thrombocytes.
The thrombocyte is the avian equivalent of the mammalian blood platelet and is involved in hemostasis through a fibrinogen-mediated process. Although fibrinogen has been implicated as a molecular bridge between activated cells during aggregation, the location of this molecule and its receptor on thrombocytes has not been characterized. Pigeon fibrinogen, isolated from plasma by precipitation with PEG-1000 and purified over Sepharose 4B, was used to study receptor-ligand interaction. Separation of pigeon fibrinogen on SDS-PAGE resulted in three peptides of molecular mass 62, 55, and 47 kDa, which were comparable to those of human fibrinogen. The role of fibrinogen and its receptor in thrombocyte function was established by turbidimetric aggregation using thrombin as an agonist under conditions requiring Ca2+ and fibrinogen. Maximum response occurred using 3 mM Ca2+ and 100 micrograms/ml fibrinogen. Fibrinogen-dependent aggregation was inhibited by an anti-GPIIb antibody, verifying a role for fibrinogen receptors in thrombocyte function. Fibrinogen-gold conjugates were used to describe receptor and ligand localization on aggregated cells. Computer reconstruction was used to verify relocalization of fibrinogen receptors following activation. Fibrinogen distribution changed from a dispersed state in preactivated cells to focal localization at points of cell contact and along pseudopods following activation. This selective positioning of fibrinogen suggests that a functional relocalization of the receptor occurs upon thrombocyte activation, and this relocation facilitates the role of fibrinogen as a molecular bridge. These studies establish similarities between the avian and the human systems and document the conserved nature of the hemostatic process.
O'Toole
E T
ET
Department of Pathology, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27157, USA.
Hantgan
R R
RR
Lewis
J C
JC
D000818:Animals; D001792:Blood Platelets / Q000737:chemistry* / Q000187:drug effects / Q000648:ultrastructure; D010856:Columbidae; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000032:analysis* / Q000737:chemistry; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D015394:Molecular Structure; D010974:Platelet Aggregation / Q000502:physiology*; D010980:Platelet Membrane Glycoproteins / Q000032:analysis*; D013917:Thrombin / Q000494:pharmacology
D010980:Platelet Membrane Glycoproteins; D005340:Fibrinogen; D013917:Thrombin