title: Difference of (Ca2+)i movements in platelets stimulated by thrombin and TRAP: the involvement of alpha(IIb)beta3-mediated TXA2 synthesis.
abstract: This study investigated the difference of [Ca2+]i movement in platelets in response to thrombin and TRAP. The involvement of alpha(IIb)beta3 in this signaling was also studied. Stimulation of platelets with thrombin at 0.03 U/ml caused platelet aggregation and a two-peak increase in [Ca2+]i. The second peak of [Ca2+]i, but not the first peak was abolished by the inhibition of platelet aggregation with alpha(IIb)beta3 antagonists or by scavenging endogenous ADP with apyrase. A cyclooxygenase inhibitor, aspirin, and a TXA2 receptor antagonist, BM 13505, also abolished the second peak of [Ca2+]i but not the first peak, although these regents did not inhibit aggregation. Under the same assay conditions, measurement of TXB2 demonstrated that alpha(IIb)beta3 antagonists and aspirin almost completely inhibited the production of TXB2. In contrast to thrombin-stimulation, TRAP caused only a single peak of [Ca2+]i even in the presence of platelet aggregation, and a high level of [Ca2+]i increase was needed for the induction of platelet aggregation. The inhibition of aggregation with alpha(IIb)beta3 antagonists had no effect on [Ca2+]i change and TXB2 production induced by TRAP. Inhibition studies using anti-GPIb antibodies suggested that GPIb may be involved in the thrombin response, but not in the TRAP. Our findings suggest that low dose thrombin causes a different [Ca2+]i response and TXA2 producing signal from TRAP. Endogenous ADP release and fibrinogen binding to alpha(IIb)beta3 are responsible for the synthesis of TXA2 which results in the induction of the second peak of [Ca2+]i in low thrombin- but not TRAP-stimulated platelets.
authors.affiliation: Department of Pharmacology, Medicinal Biology Research Laboratories, Fujisawa Pharmaceutical Co., Ltd., Osaka, Japan. toshiaki_aoki@po.fujisawa.co.jp
authors.forename: T
authors.initials: T
authors.lastname: Aoki
authors.forename: Y
authors.initials: Y
authors.lastname: Tomiyama
authors.forename: S
authors.initials: S
authors.lastname: Honda
authors.forename: K
authors.initials: K
authors.lastname: Senzaki
authors.forename: A
authors.initials: A
authors.lastname: Tanaka
authors.forename: M
authors.initials: M
authors.lastname: Okubo
authors.forename: F
authors.initials: F
authors.lastname: Takahashi
authors.forename: H
authors.initials: H
authors.lastname: Takasugi
authors.forename: J
authors.initials: J
authors.lastname: Seki
mesh_terms: D000244:Adenosine Diphosphate / Q000037:antagonists & inhibitors / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D001081:Apyrase / Q000494:pharmacology; D001241:Aspirin / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D004151:Dipeptides / Q000494:pharmacology; D005340:Fibrinogen / Q000378:metabolism; D006801:Humans; D017136:Ion Transport; D010446:Peptide Fragments / Q000494:pharmacology*; D010648:Phenylacetates / Q000494:pharmacology; D015539:Platelet Activation / Q000187:drug effects; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000037:antagonists & inhibitors / Q000502:physiology*; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000037:antagonists & inhibitors; D018179:Receptors, Thrombin / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013449:Sulfonamides / Q000494:pharmacology; D013917:Thrombin / Q000494:pharmacology*; D013928:Thromboxane A2 / Q000096:biosynthesis*; D013929:Thromboxane B2 / Q000032:analysis; D000077466:Tirofiban; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D016861:Cyclooxygenase Inhibitors; D004151:Dipeptides; D010446:Peptide Fragments; D010648:Phenylacetates; D010975:Platelet Aggregation Inhibitors; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019038:Platelet Glycoprotein GPIb-IX Complex; D018179:Receptors, Thrombin; D013449:Sulfonamides; C082835:thrombin receptor peptide (42-47); C103087:((4-(4-amidinophenoxy)butanoyl)aspartyl)valine; D014443:Tyrosine; D013929:Thromboxane B2; D013928:Thromboxane A2; D000244:Adenosine Diphosphate; D005340:Fibrinogen; D013917:Thrombin; D001081:Apyrase; D000077466:Tirofiban; D001241:Aspirin; C051762:daltroban; D002118:Calcium