Src family kinases are essential for primary aggregation by G(i) -coupled receptors.
BACKGROUND
Adrenaline stimulates biphasic aggregation in plasma through the G(i) -coupled α(2A) -adrenoreceptor. In the present study, we demonstrate that both primary and secondary wave aggregation induced by adrenaline in plasma is blocked by two structurally distinct inhibitors of Src family kinases, dasatinib and PD0173952.


RESULTS
In contrast, primary aggregation is partially inhibited or unaffected in the presence of inhibitors of cyclo-oxygenase, phosphoinositide (PI) 3-kinases, and P2Y(1) and P2Y(12) ADP receptors, although secondary aggregation is abolished. The ability of adrenaline to inhibit adenylyl cyclase and to synergize with platelet agonists in mediating platelet activation in plasma is retained in the presence of Src family kinase inhibition. Moreover, adrenaline does not activate Src family kinases, as determined by western blotting of their regulatory tyrosines, suggesting that constitutive signaling from Src family kinases may underlie their role in activation. Adrenaline is widely used in clinical laboratories for investigation of patients with suspected bleeding disorders. In a group of 90 unrelated patients with a clinically diagnosed platelet bleeding disorder, we identified four who did not exhibit primary wave aggregation in response to adrenaline, although the catecholamine potentiated the response to other agonists, and five who failed to undergo secondary wave aggregation. In contrast, adrenaline stimulated biphasic aggregation in 60 controls. All of the patients with a defective response to adrenaline had impaired ADP-induced platelet activation.


CONCLUSIONS
The present results indicate a previously unappreciated role for Src family kinases in mediating G(i) signaling in plasma, and demonstrate heterogeneity in response to adrenaline in patients with a clinically diagnosed platelet disorder.
Nash
C A
CA
Centre for Cardiovascular Sciences, Institute of Biomedical Research, College of Medical and Dental Sciences, University of Birmingham, UK. can562@bham.ac.uk
Séverin
S
S
Dawood
B B
BB
Makris
M
M
Mumford
A
A
Wilde
J
J
Senis
Y A
YA
Watson
S P
SP
D001372:Aza Compounds / Q000494:pharmacology; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D000069439:Dasatinib; D004837:Epinephrine / Q000378:metabolism; D019206:GTP-Binding Protein alpha Subunits, Gi-Go / Q000378:metabolism*; D006801:Humans; D009025:Morpholines / Q000494:pharmacology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D015539:Platelet Activation; D053657:Platelet-Rich Plasma / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D058925:Receptors, Purinergic P2Y12 / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology; D014443:Tyrosine / Q000737:chemistry; D019061:src-Family Kinases / Q000502:physiology*
C511991:8-(2,6-dichlorophenyl)-10-methyl-3-((4-morpholin-4-ylphenyl)amino)-2,4,10-triazabicyclo(4.4.0)deca-1,3,5,7-tetraen-9-one; D001372:Aza Compounds; D009025:Morpholines; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D058925:Receptors, Purinergic P2Y12; D013844:Thiazoles; D014443:Tyrosine; D000242:Cyclic AMP; D019869:Phosphatidylinositol 3-Kinases; D019061:src-Family Kinases; D019206:GTP-Binding Protein alpha Subunits, Gi-Go; D000069439:Dasatinib; D004837:Epinephrine