Respective contributions of single and compound granule fusion to secretion by activated platelets.
Although granule secretion is pivotal in many platelet responses, the fusion routes of α and δ granule release remain uncertain. We used a 3D reconstruction approach based on electron microscopy to visualize the spatial organization of granules in unstimulated and activated platelets. Two modes of exocytosis were identified: a single mode that leads to release of the contents of individual granules and a compound mode that leads to the formation of granule-to-granule fusion, resulting in the formation of large multigranular compartments. Both modes occur during the course of platelet secretion. Single fusion events are more visible at lower levels of stimulation and early time points, whereas large multigranular compartments are present at higher levels of agonist and at later time points. Although α granules released their contents through both modes of exocytosis, δ granules underwent only single exocytosis. To define the underlying molecular mechanisms, we examined platelets from vesicle-associated membrane protein 8 (VAMP8) null mice. After weak stimulation, compound exocytosis was abolished and single exocytosis decreased in VAMP8 null platelets. Higher concentrations of thrombin bypassed the VAMP8 requirement, indicating that this isoform is a key but not a required factor for single and/or compound exocytosis. Concerning the biological relevance of our findings, compound exocytosis was observed in thrombi formed after severe laser injury of the vessel wall with thrombin generation. After superficial injury without thrombin generation, no multigranular compartments were detected. Our studies suggest that platelets use both modes of membrane fusion to control the extent of agonist-induced exocytosis.
INSERM, Unité Mixte de Recherche-S949.|Etablissement Français du Sang.|Université de Strasbourg, Faculté de Médecine, and.|Fédération de Médecine Translationnelle, Strasbourg, France; and.
Anita
A
Eckly
INSERM, Unité Mixte de Recherche-S949.|Etablissement Français du Sang.|Université de Strasbourg, Faculté de Médecine, and.|Fédération de Médecine Translationnelle, Strasbourg, France; and.
Jean-Yves
JY
Rinckel
INSERM, Unité Mixte de Recherche-S949.|Etablissement Français du Sang.|Université de Strasbourg, Faculté de Médecine, and.|Fédération de Médecine Translationnelle, Strasbourg, France; and.
Fabienne
F
Proamer
INSERM, Unité Mixte de Recherche-S949.|Etablissement Français du Sang.|Université de Strasbourg, Faculté de Médecine, and.|Fédération de Médecine Translationnelle, Strasbourg, France; and.
Neslihan
N
Ulas
Department of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY.
Smita
S
Joshi
Department of Molecular and Cellular Biochemistry, University of Kentucky College of Medicine, Lexington, KY.
Sidney W
SW
Whiteheart
INSERM, Unité Mixte de Recherche-S949.|Etablissement Français du Sang.|Université de Strasbourg, Faculté de Médecine, and.|Fédération de Médecine Translationnelle, Strasbourg, France; and.
Christian
C
Gachet
D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D005089:Exocytosis; D051379:Mice; D008817:Mice, Mutant Strains; D015539:Platelet Activation; D050683:R-SNARE Proteins / Q000235:genetics / Q000378:metabolism*; D022142:Secretory Vesicles / Q000235:genetics / Q000378:metabolism*
D050683:R-SNARE Proteins; C491831:Vamp8 protein, mouse