Inhibition of caspases but not of calpains temporarily protect against C2-ceramide-induced death of CAD cells.
Evidence has implicated apoptosis as a mechanism underlying cell death in diverse neurodegenerative diseases including Parkinson's disease (PD). Endogenous agents such as TNF-alpha, INF-gamma, IL-1beta and others stress signals activate the sphingomyelin pathway increasing ceramide levels. Ceramide triggers apoptotic pathways while inhibiting survival signalling, and is involved in the regulation of intracellular Ca(2+) homeostasis and compartmentalisation. The contribution of caspases in neuronal apoptosis has been highlighted by the increased survival exerted by caspase inhibition, but the involvement of calpains during neuronal apoptosis and the potential benefit of their inhibition is still controversial. In the present paper, we have analysed the contribution of caspases and calpains to cell death of CAD cells, a catecholaminergic cell line of mesencephalic origin, following C2-ceramide exposure. Ceramide caused CAD cell death by a dose and time dependant mechanism. 25microM of C2-ceramide caused apoptosis. Analysis of activation of caspases and calpains by differential cleavage of alpha-fodrin showed that although calpains are activated before caspases following C2-ceramide exposure, only caspase inhibition increased cell survival. These results demonstrate the activation of caspases and calpains in C2-ceramide-induced cell death, and support the role of caspase inhibition as a neuroprotective strategy and a plausible therapeutic approach to decrease catecholaminergic cell death.
Arboleda
Gonzalo
G
Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester M13 9PT, UK. gharboledab@unal.edu.co
Waters
Catherine
C
Gibson
Rosemary
R
D000704:Analysis of Variance; D000818:Animals; D002154:Calpain / Q000378:metabolism*; D020169:Caspases / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D004151:Dipeptides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004791:Enzyme Inhibitors / Q000633:toxicity; D051379:Mice; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000378:metabolism*; D013110:Sphingosine / Q000031:analogs & derivatives / Q000633:toxicity; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors
D004151:Dipeptides; D004791:Enzyme Inhibitors; C064769:N-acetylsphingosine; D018696:Neuroprotective Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; C071482:calpeptin; D002154:Calpain; D020169:Caspases; C022616:thiazolyl blue; D013110:Sphingosine