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Characteristics of endogenous γ-aminobutyric acid (GABA) in human platelets: functional studies of a novel collagen glycoprotein VI inhibitor

Journal of Molecular Medicine volume 92, pages 603–614 (2014)Cite this article

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/106 cells. GABA (0.5 μM) specifically inhibited collagen-induced platelet activation accompanied by [Ca2+]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 α2β1 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.

Key messages

  • 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.

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Acknowledgments

We wish to thank Prof. Tur-Fu Huang in the Department of Pharmacology (College of Medicine, National Taiwan University) for providing convulxin and aggretin for our experiments. This work was supported by grants from the National Science Council, Taiwan (NSC97-2320-B-038-016-MY3, NSC100-2320-B-038-021-MY3, NSC102-2320-B-341-001-MY3, and NSC102-2811-B-038-026), Taipei Medical University Hospital (100 TMU-TMUH-03), Shin Kong Wu Ho-Su Memorial Hospital (SKH-8302-102-NDR-04 and SKH-8302-103-NDR-05), and Cathay General Hospital (100CGH-TMU-12).

Conflict of interest

The authors declare no competing financial interests.

Author information

Affiliations

  1. Central Laboratory, Shin-Kong Wu Ho-Su Memorial Hospital, 95 Wen-Chang Rd., Shih-Lin, Taipei, 11101, Taiwan

    Kuan-Hung Lin

  2. Graduate Institute of Medical Sciences and Department of Pharmacology, College of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei, 11031, Taiwan

    Kuan-Hung Lin, Wan-Jung Lu, Shwu-Huey Wang, Duen-Suey Chou, Yung-Chen Chiang & Joen-Rong Sheu

  3. Department of Anatomy, College of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei, 11031, Taiwan

    Tsorng-Harn Fong

  4. Department of Cardiology, Cathay General Hospital, Taipei, 11687, Taiwan

    Chao-Chien Chang

  5. Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, 250 Wu-Hsing St., Taipei, 11031, Taiwan

    Nen-Chung Chang

  6. School of Nutrition and Health Sciences, Taipei Medical University, 250 Wu-Hsing St., Taipei, 11031, Taiwan

    Wan-Jung Lu & Shih-Yi Huang

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  1. Kuan-Hung Lin
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  2. Wan-Jung Lu
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Correspondence to Joen-Rong Sheu.

Additional information

K.H. Lin and W.J. Lu contributed equally to this manuscript.

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Lin, KH., Lu, WJ., Wang, SH. et al. Characteristics of endogenous γ-aminobutyric acid (GABA) in human platelets: functional studies of a novel collagen glycoprotein VI inhibitor. J Mol Med 92, 603–614 (2014). https://doi.org/10.1007/s00109-014-1140-7

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  • DOI: https://doi.org/10.1007/s00109-014-1140-7

Keywords

  • γ-Aminobutyric acid
  • Collagen
  • Convulxin
  • Glycoprotein VI antagonist
  • Platelet activation