title: Characteristics of endogenous γ-aminobutyric acid (GABA) in human platelets: functional studies of a novel collagen glycoprotein VI inhibitor.
abstract: gamma-Aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system, and it also appears in peripheral tissues. Platelets are anuclear blood cells that play a central role in hemostatic processes. Although platelets possess a GABA uptake system, the functional activity of GABA in platelets has remained unclear. We determined that GABA is abundantly distributed in the platelets at a concentration of approximately 1.03 ng/10(6) cells. GABA (0.5 μM) specifically inhibited collagen-induced platelet activation accompanied by [Ca(2+)]i mobilization, phospholipase Cγ2, protein kinase C, Akt phosphorylation, and hydroxyl radical formation. In addition, GABA interfered with fluorescein isothiocyanate-collagen binding to platelet membranes and produced a concentration-dependent shift in the collagen concentration-response curve and a Schild plot slope of -0.96 ± 0.11, indicating competitive inhibition. Platelet activation induced by convulxin, a glycoprotein VI agonist, was inhibited by GABA, whereas activation induced by the integrin α(2)β(1) agonist, aggretin, was not. Immunoprecipitation and surface plasmon resonance revealed that GABA binds directly to glycoprotein VI in human platelets with equilibrium dissociation (binding) constant (K(D)) of 41.4 nM. The closure time of whole blood and the occlusion time of platelet plug formation were significantly prolonged by GABA in vivo. In this study, GABA is a specific inhibitor of collagen glycoprotein VI and may be involved in an endogenous negative feedback mechanism for platelet activation. Thus, GABA may represent a potential target for the development of novel interventions for the treatment of cardiovascular diseases associated with platelet activation, such as stroke and myocardial infarction.
  
  
  CONCLUSIONS
  GABA is abundantly distributed in the platelets. GABA inhibited platelet activation stimulated by convulxin. GABA directly associated with glycoprotein VI in platelet membrane. GABA prolonged the closure time of whole blood and the occlusion time of platelet plug formation in vivo.
authors.affiliation: Central Laboratory, Shin-Kong Wu Ho-Su Memorial Hospital, 95 Wen-Chang Rd., Shih-Lin, Taipei, 11101, Taiwan.
authors.forename: Kuan-Hung
authors.initials: KH
authors.lastname: Lin
authors.forename: Wan-Jung
authors.initials: WJ
authors.lastname: Lu
authors.forename: Shwu-Huey
authors.initials: SH
authors.lastname: Wang
authors.forename: Tsorng-Harn
authors.initials: TH
authors.lastname: Fong
authors.forename: Duen-Suey
authors.initials: DS
authors.lastname: Chou
authors.forename: Chao-Chien
authors.initials: CC
authors.lastname: Chang
authors.forename: Nen-Chung
authors.initials: NC
authors.lastname: Chang
authors.forename: Yung-Chen
authors.initials: YC
authors.lastname: Chiang
authors.forename: Shih-Yi
authors.initials: SY
authors.lastname: Huang
authors.forename: Joen-Rong
authors.initials: JR
authors.lastname: Sheu
mesh_terms: D001792:Blood Platelets / Q000187:drug effects* / Q000378:metabolism*; D003435:Crotalid Venoms / Q000494:pharmacology; D005434:Flow Cytometry; D006801:Humans; D047468:Immunoprecipitation; D037181:Lectins, C-Type; D015539:Platelet Activation / Q000187:drug effects*; D010980:Platelet Membrane Glycoproteins / Q000819:agonists; D020349:Surface Plasmon Resonance; D053719:Tandem Mass Spectrometry; D005680:gamma-Aminobutyric Acid / Q000378:metabolism* / Q000494:pharmacology*
chemical_list: D003435:Crotalid Venoms; D037181:Lectins, C-Type; D010980:Platelet Membrane Glycoproteins; C098341:platelet membrane glycoprotein VI; C028890:convulxin; D005680:gamma-Aminobutyric Acid