C2-ceramide mediates cerebellar granule cells apoptosis by activation of caspases-2, -9, and -3.
Ceramide is able to induce the apoptotic death of cerebellar granule cells (CGC) in culture. However, previous reports did not agree on whether ceramide-induced apoptosis of CGC requires caspase activation. Here we have shown that addition of C2-ceramide is able to produce extensive death of cultured CGC, which is associated with chromatin condensation, ladder-like DNA fragmentation, and activation of caspases. Our results show that C2-ceramide activates caspases-3, -9, and -2 but not caspases-1 and -8. Caspase-9 activation was associated with cytochrome c release from mitochondria toward the cytosol and was followed by activation of caspases-2 and -3. PARP proteolysis was also observed after caspase-3 and -2 activation. The involvement of caspases-9, -3, and -2 in ceramide-mediated apoptotic death of CGC was further supported by the use of specific inhibitors.
Miñano
Alfredo
A
Institut de Neurociencies and Department de Bioquímica i Biología Molecular, Universitat Autònoma de Barcelona, Barcelona, Spain.
Caballero-Benítez
Andrea
A
Lluch
Mónica
M
Morán
Julio
J
Rodríguez-Alvarez
José
J
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D053143:Caspase 2 / Q000235:genetics / Q000378:metabolism*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism*; D053453:Caspase 9 / Q000235:genetics / Q000378:metabolism*; D061945:Caspase Inhibitors; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002531:Cerebellum / Q000187:drug effects / Q000201:enzymology*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D051381:Rats; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000502:physiology
D061945:Caspase Inhibitors; D004791:Enzyme Inhibitors; C064769:N-acetylsphingosine; D053143:Caspase 2; D053148:Caspase 3; D053453:Caspase 9; D013110:Sphingosine