Pharmacological Characterization of the Mechanisms Involved in Delayed Calcium Deregulation in SH-SY5Y Cells Challenged with Methadone.

Filed under: Methadone Clinics

Int J Cell Biol. 2012; 2012: 642482
Perez-Alvarez S, Solesio ME, Cuenca-Lopez MD, Melero-Fernández de Mera RM, Villalobos C, Kmita H, Galindo MF, Jordán J

Previously, we have shown that SH-SY5Y cells exposed to high concentrations of methadone died due to a necrotic-like cell death mechanism related to delayed calcium deregulation (DCD). In this study, we show that, in terms of their Ca(2+) responses to 0.5?mM methadone, SH-SY5Y cells can be pooled into four different groups. In a broad pharmacological survey, the relevance of different Ca(2+)-related mechanisms on methadone-induced DCD was investigated including extracellular calcium, L-type Ca(2+) channels, ?-opioid receptor, mitochondrial inner membrane potential, mitochondrial ATP synthesis, mitochondrial Ca(2+)/2Na(+)-exchanger, reactive oxygen species, and mitochondrial permeability transition. Only those compounds targeting mitochondria such as oligomycin, FCCP, CGP 37157, and cyclosporine A were able to amend methadone-induced Ca(2+) dyshomeostasis suggesting that methadone induces DCD by modulating the ability of mitochondria to handle Ca(2+). Consistently, mitochondria became dramatically shorter and rounder in the presence of methadone. Furthermore, analysis of oxygen uptake by isolated rat liver mitochondria suggested that methadone affected mitochondrial Ca(2+) uptake in a respiratory substrate-dependent way. We conclude that methadone causes failure of intracellular Ca(2+) homeostasis, and this effect is associated with morphological and functional changes of mitochondria. Likely, this mechanism contributes to degenerative side effects associated with methadone treatment.
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