Journal Article

Dissecting the biochemical architecture and morphological release pathways of the human platelet extracellular vesiculome

Wu, Yong, Alayash, Abdu, Strader, Michael B., Diduch, Lukas L., Simak, Jan, De Paoli, Silvia H., Zheng, Jiwen, Patel, Mehulkumar, Tarandovskiy, Ivan D., Tegegn, Tseday Z., Ovanesov, Mikhail V., Elhelu, Oumsalama K.

2018

Overview

Platelet extracellular vesicles (PEVs) have emerged as potential mediators in intercellular communication. PEVs exhibit several activities with pathophysiological importance and may serve as diagnostic biomarkers. Here, imaging and analytical techniques were employed to unveil morphological pathways of the release, structure, composition, and surface properties of PEVs derived from human platelets (PLTs) activated with the thrombin receptor activating peptide (TRAP). Based on extensive electron microscopy analysis, we propose four morphological pathways for PEVs release from TRAP-activated PLTs: (1) plasma membrane budding, (2) extrusion of multivesicular α-granules and cytoplasmic vacuoles, (3) plasma membrane blistering and (4) “pearling” of PLT pseudopodia. The PLT extracellular vesiculome encompasses ectosomes, exosomes, free mitochondria, mitochondria-containing vesicles, “podiasomes” and PLT “ghosts”. Interestingly, a flow cytometry showed a population of TOM20 + LC3 + PEVs, likely products of platelet mitophagy. We found that lipidomic and proteomic profiles were different between the small PEV (S-PEVs; mean diameter 103 nm) and the large vesicle (L-PEVs; mean diameter 350 nm) fractions separated by differential centrifugation. In addition, the majority of PEVs released by activated PLTs was composed of S-PEVs which have markedly higher thrombin generation activity per unit of PEV surface area compared to L-PEVs, and contribute approximately 60% of the PLT vesiculome procoagulant potency.

• Publisher: Springer International Publishing,Springer Nature B.V
• Subject: Biochemistry / Biomarkers / Biomedical and Life Sciences / Biomedicine / Blistering / blood platelets / Blood Platelets - cytology / Blood Platelets - metabolism / Cell Biology / cell communication / Cell Membrane - metabolism / Cell signaling / Cell-Derived Microparticles - metabolism / Centrifugation / Chemokines - metabolism / Cytokines - metabolism / Cytometry / Diagnostic systems / Diameters / Electron microscopy / Exosomes / Exosomes - metabolism / Extrusion / Flow cytometry / Fractions / Ghosts / Humans / image analysis / Life Sciences / Lipids - analysis / Membrane Transport Proteins - metabolism / Membranes / Microscopy, Confocal / Microscopy, Electron, Transmission / Microtubule-Associated Proteins - metabolism / Mitochondria / Mitochondrial Precursor Protein Import Complex Proteins / Mitophagy / Morphology / Original / Original Article / Particle Size / Peptide Fragments - metabolism / peptides / plasma membrane / Platelets / Proteomics / Pseudopodia / Receptors, Cell Surface - metabolism / SNARE Proteins - metabolism / surface area / Surface properties / Thrombin / Thrombin - metabolism / Vacuoles / Vesicles