Factors that affect the size of platelet aggregates in epinephrine-induced activation: a study using the particle counting method based upon light scattering.
Platelet aggregate size was determined with a newly-developed platelet aggregometer, PA-100 (KOWA), which can quantitatively evaluate the size and number of platelet aggregates by means of the particle counting method based upon light scattering. Epinephrine-induced platelet aggregation consists of two phases, the former characterized by the formation of small-sized aggregates (less than 100 cells), which is followed by the phase of large aggregate formation with concomitant decrease in the number of small aggregates. These findings suggest that small aggregates fuse to form large aggregates. Effects of various inhibitors and antibodies directed against platelet membrane glycoproteins were evaluated on the size of platelet aggregates induced by epinephrine. Cyclooxygenase inhibitors, thromboxane A2 receptor antagonists, and a Na+/H+ exchanger inhibitor (ethylisopropylamiloride) inhibited the formation of large aggregates (more than 100 cells) but not that of small aggregates. Cytochalasin B, which interferes with microfilaments, suppressed large aggregate formation, whereas taxol, which reacts with microtubules, had no effects. Anti-GPIIb/IIIa monoclonal antibody (MoAb) inhibited both the formation of small and large platelet aggregates, while antibodies directed against GPIb, thrombospondin, P-selectin, or PECAM-1 had no effects on platelet aggregate formation. These findings, taken together, suggest that intracellular alkalinization, thromboxane A2 formation and microfilament rearrangement are prerequisites for large platelet aggregate formation. GPIIb/IIIa is involved in the formation of small as well as large aggregates, but a membrane glycoprotein(s) responsible for the transition of small aggregates into large aggregates awaits to be determined.
Department of Clinical and Laboratory Medicine, Yamanashi Medical University, Tamaho, Nakakoma, Japan.
K
K
Satoh
Y
Y
Ozaki
R
R
Qi
L
L
Yang
N
N
Asazuma
Y
Y
Yatomi
S
S
Kume
D016718:Arachidonic Acid / Q000037:antagonists & inhibitors; D004837:Epinephrine / Q000494:pharmacology*; D006801:Humans; D008027:Light; D010316:Particle Size; D010974:Platelet Aggregation / Q000187:drug effects*; D010979:Platelet Function Tests / Q000379:methods; D010980:Platelet Membrane Glycoproteins / Q000037:antagonists & inhibitors; D012542:Scattering, Radiation; D017923:Sodium-Hydrogen Exchangers / Q000037:antagonists & inhibitors
D010980:Platelet Membrane Glycoproteins; D017923:Sodium-Hydrogen Exchangers; D016718:Arachidonic Acid; D004837:Epinephrine