Role for the guanine nucleotide exchange factor phosphatidylinositol-3,4,5-trisphosphate-dependent rac exchanger 1 in platelet secretion and aggregation

Arterioscler Thromb Vasc Biol. 2012 Mar;32(3):768-77. doi: 10.1161/ATVBAHA.111.243675. Epub 2011 Dec 29.

Abstract

Objective: Recent studies have shown a role for Rac1 in regulating platelet functions, but how Rac1 is activated in platelets remains unclear. Phosphatidylinositol-3,4,5-trisphosphate-dependent Rac exchanger 1 (P-Rex1) was originally identified in neutrophils that regulates phagocyte functions. We sought to examine whether P-Rex1 plays a role in platelet activation.

Methods and results: Western blotting showed P-Rex1 expression in mouse and human platelets. Mice lacking P-Rex1 exhibited prolonged bleeding time and increased rebleeding. When challenged with low doses of the G protein-coupled receptor (GPCR) agonists U46619 and thrombin, P-Rex1-/- platelets displayed significantly reduced secretion and aggregation compared with wild-type platelets. Increasing the concentration of these agonists could overcome the defect. Platelet aggregation induced by collagen, a non-GPCR agonist, was also compromised in the absence of P-Rex1. Along with these phenotypic changes were impaired Rac1 activation; reduced ATP secretion; and decreased phosphorylation of Akt, c-Jun N-terminal kinase, and p38 mitogen-activated protein kinase in P-Rex1-/- platelets on agonist stimulation.

Conclusion: These results demonstrate for the first time the presence of P-Rex1 in platelets as well as its role in platelet secretion and aggregation induced by low-dose agonists for GPCR and by collagen.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / pharmacology
  • Adenosine Triphosphate / metabolism
  • Animals
  • Bleeding Time
  • Blood Platelets / drug effects
  • Blood Platelets / metabolism*
  • Blotting, Western
  • Collagen / pharmacology
  • Cytoplasmic Granules / metabolism
  • Dose-Response Relationship, Drug
  • Enzyme Activation
  • Female
  • Genotype
  • Guanine Nucleotide Exchange Factors / deficiency
  • Guanine Nucleotide Exchange Factors / genetics
  • Guanine Nucleotide Exchange Factors / metabolism*
  • HEK293 Cells
  • Hemostasis / genetics
  • Humans
  • JNK Mitogen-Activated Protein Kinases / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Phenotype
  • Phosphorylation
  • Platelet Aggregation* / drug effects
  • Proto-Oncogene Proteins c-akt / metabolism
  • Receptors, G-Protein-Coupled / agonists
  • Receptors, G-Protein-Coupled / metabolism
  • Signal Transduction
  • Thrombin / metabolism
  • Transfection
  • p38 Mitogen-Activated Protein Kinases / metabolism
  • rac1 GTP-Binding Protein / metabolism

Substances

  • Guanine Nucleotide Exchange Factors
  • PREX1 protein, human
  • PREX1 protein, mouse
  • Receptors, G-Protein-Coupled
  • 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid
  • Adenosine Triphosphate
  • Collagen
  • Proto-Oncogene Proteins c-akt
  • JNK Mitogen-Activated Protein Kinases
  • p38 Mitogen-Activated Protein Kinases
  • Thrombin
  • rac1 GTP-Binding Protein