Inhibition of mitochondrial cytochrome c oxidase potentiates Aβ-induced ER stress and cell death in cortical neurons

• Published in: Molecular and cellular neuroscience Vol. 52; pp. 1 - 8
• Main Authors: Costa, Rui O., Ferreiro, Elisabete, Oliveira, Catarina R., Pereira, Cláudia M.F.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.2013
• Subjects: Alzheimer's disease; Amyloid beta-Peptides - metabolism; Amyloid beta-Peptides - toxicity; Amyloid-β; Animals; Apoptosis; Apoptosis - physiology; beta -Amyloid; Blotting, Western; Calcium; Cell activation; Cell death; Cell survival; Cerebral Cortex - metabolism; Communication; Cortex; Cytochrome c oxidase; Data processing; Electron transport chain; Electron Transport Complex IV - metabolism; Endoplasmic reticulum; Endoplasmic Reticulum Stress - physiology; Mitochondria; Mitochondria - metabolism; Neurodegenerative diseases; Neurons; Neurons - metabolism; potassium cyanide; Protein folding; Rats; Rats, Wistar; Ryanodine receptors; Stress; thapsigargin; Cytochrome c oxidase; Electron transport chain; Mitochondria; Amyloid-β; Alzheimer's disease; Endoplasmic reticulum
• ISSN: 1044-7431; 1095-9327
• DOI: 10.1016/j.mcn.2012.09.005

Previously we reported that amyloid-β (Aβ) leads to endoplasmic reticulum (ER) stress in cultured cortical neurons and that ER-mitochondria Ca2+ transfer is involved in Aβ-induced apoptotic neuronal cell death. In cybrid cells which recreate the defect in mitochondrial cytochrome c oxidase (COX) activity observed in platelets from Alzheimer's disease (AD) patients, we have shown that mitochondrial dysfunction affects the ER stress response triggered by Aβ. Here, we further investigated the impact of COX inhibition on Aβ-induced ER dysfunction using a neuronal model. Primary cultures of cortical neurons were challenged with toxic concentrations of Aβ upon chemical inhibition of COX with potassium cyanide (KCN). ER Ca2+ homeostasis was evaluated under these conditions, together with the levels of ER stress markers, namely the chaperone GRP78 and XBP-1, a mediator of the ER unfolded protein response (UPR). We demonstrated that COX inhibition potentiates the Aβ-induced depletion of ER Ca2+ content. KCN pre-treatment was also shown to enhance the rise of cytosolic Ca2+ levels triggered by Aβ and thapsigargin, a widely used ER stressor. This effect was reverted in the presence of dantrolene, an inhibitor of ER Ca2+ release through ryanodine receptors. Similarly, the increase in GRP78 and XBP-1 protein levels was shown to be higher in neurons treated with Aβ or thapsigargin in the presence of KCN in comparison with levels determined in neurons treated with the neurotoxins alone. Although the decrease in cell survival, the activation of caspase-9- and -3-mediated apoptotic cell death observed in Aβ- and thapsigargin-treated neurons were also potentiated by KCN, this effect is less pronounced than that observed in Ca2+ signalling and UPR. Furthermore, in neurons treated with Aβ, the potentiating effect of the COX inhibitor in cell survival and death was not prevented by dantrolene. These results show that inhibition of mitochondrial COX activity potentiates Aβ-induced ER dysfunction and, to a less extent, neuronal cell death. Furthermore, data supports that the effect of impaired COX on Aβ-induced cell death occurs independently of Ca2+ release through ER ryanodine receptors. Together, our data demonstrate that mitochondria dysfunction in AD enhances the neuronal susceptibility to toxic insults, namely to Aβ-induced ER stress, and strongly suggest that the close communication between ER and mitochondria can be a valuable future therapeutic target in AD.