title: Mitochondrial control of platelet apoptosis: effect of cyclosporin A, an inhibitor of the mitochondrial permeability transition pore.
abstract: The role of the mitochondrial permeability transition pore (MPTP) in apoptosis of nucleated cells is well documented. In contrast, the role of MPTP in apoptosis of anucleated platelets is largely unknown. The aim of this study was to elucidate the contribution of MPTP in the control of different manifestations of platelet apoptosis by analyzing the effect of cyclosporin A (CsA), a potent inhibitor of MPTP formation. Using flow cytometry, we studied the effect of pretreatment of platelets with CsA on apoptotic responses in human platelets stimulated with calcium ionophore A23187. We found that CsA inhibited A23187-stimulated platelet apoptosis, completely preventing (i) depolarization of mitochondrial inner membrane potential (DeltaPsim), (ii) activation of cytosolic apoptosis executioner caspase-3, (iii) platelet shrinkage, and (iv) fragmentation of platelets to microparticles, but (v) only partially (approximately 25%), inhibiting phosphatidylserine (PS) exposure on the platelet surface. This study shows that MPTP formation is upstream of DeltaPsim depolarization, caspase-3 activation, platelet shrinkage and microparticle formation, and stringently controls these apoptotic events in A23187-stimulated platelets but is less involved in PS externalization. These data also indicate that CsA may rescue platelets from apoptosis, preventing caspase-3 activation and inhibiting the terminal cellular manifestations of platelet apoptosis, such as platelet shrinkage and degradation to microparticles. Furthermore, the results suggest a novel potentially useful application of CsA as an inhibitor of platelet demise through apoptosis in thrombocytopenias associated with enhanced platelet apoptosis.
authors.affiliation: Division of Transfusion Medicine, Department of Laboratory Medicine, The Keenan Research Centre in the Li Ka Shing Knowledge Institute of St. Michael's Hospital, ON, Canada. leytinv@smh.toronto.on.ca
authors.forename: Valery
authors.initials: V
authors.lastname: Leytin
authors.forename: David J
authors.initials: DJ
authors.lastname: Allen
authors.forename: Asuman
authors.initials: A
authors.lastname: Mutlu
authors.forename: Armen V
authors.initials: AV
authors.lastname: Gyulkhandanyan
authors.forename: Sergiy
authors.initials: S
authors.lastname: Mykhaylov
authors.forename: John
authors.initials: J
authors.lastname: Freedman
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D001792:Blood Platelets / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D000001:Calcimycin / Q000494:pharmacology; D053148:Caspase 3 / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology*; D004789:Enzyme Activation; D006801:Humans; D066298:In Vitro Techniques; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects / Q000502:physiology*; D008928:Mitochondria / Q000502:physiology; D033681:Mitochondrial Membrane Transport Proteins / Q000037:antagonists & inhibitors* / Q000502:physiology; D000083162:Mitochondrial Permeability Transition Pore
chemical_list: D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D000001:Calcimycin; D016572:Cyclosporine; D053148:Caspase 3