Dynamin 2 is required for GPVI signaling and platelet hemostatic function in mice

• Published in: Haematologica (Roma) 2020-05, Vol.105 (5), p.1414-1423
• Main Authors: Eaton, Nathan, Drew, Caleb, Wieser, Jon, Munday, Adam D, Falet, Hervé
• Format: Article
• Language: English
• Subjects: Animals; Blood Platelets; Dynamin II - genetics; Dynamin II - pharmacology; Hemostasis; Hemostatics - pharmacology; Mice; Platelet Activation; Platelet Membrane Glycoproteins - genetics
• ISSN: 0390-6078; 1592-8721
• DOI: 10.3324/haematol.2019.218644

Receptor-mediated endocytosis, which contributes to a wide range of cellular functions, including receptor signaling, cell adhesion, and migration, requires endocytic vesicle release by the large GTPase dynamin 2. Here, the role of dynamin 2 was investigated in platelet hemostatic function using both pharmacological and genetic approaches. Pf4-Cre ( ) mice specifically lacking dynamin 2 within the platelet lineage developed severe thrombocytopenia and bleeding diathesis and platelets adhered poorly to collagen under arterial shear rates. Signaling the collagen receptor GPVI was impaired in platelets treated with the dynamin GTPase inhibitor dynasore, as evidenced by poor protein tyrosine phosphorylation, including that of the proximal tyrosine kinase Lyn on its activating tyrosine 396 residue. Platelet stimulation GPVI resulted in a slight decrease in GPVI, which was maintained by dynasore treatment. Dynasore-treated platelets had attenuated function when stimulated GPVI, as evidenced by reduced GPIbα downregulation, α-granule release, integrin αIIbβ3 activation, and spreading onto immobilized fibrinogen. By contrast, responses to the G-protein coupled receptor agonist thrombin were minimally affected by dynasore treatment. GPVI expression was severely reduced in platelets, which were dysfunctional in response to stimulation GPVI, and to a lesser extent to thrombin. platelets lacked fibrinogen in their α-granules, but retained von Willebrand factor. Taken together, the data show that dynamin 2 plays a proximal role in signaling the collagen receptor GPVI and is required for fibrinogen uptake and normal platelet hemostatic function.