Cyanide Enhancement of Dopamine-Induced Apoptosis in Mesencephalic Cells Involves Mitochondrial Dysfunction and Oxidative Stress

• Published in: Neurotoxicology (Park Forest South) 2003-06, Vol.24 (3), p.333-342
• Main Authors: Jones, Douglas C, Prabhakaran, Krishnan, Li, Li, Gunasekar, Palur G, Shou, Yan, Borowitz, Joseph L, Isom, Gary E
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Apoptosis - physiology; Biological and medical sciences; Caspases; Cells, Cultured; Chemical and industrial products toxicology. Toxic occupational diseases; Cyanide; Cyanides - pharmacology; Dopamine; Dopamine - pharmacology; Medical sciences; Membrane Potentials - drug effects; Membrane Potentials - physiology; Mesencephalon - cytology; Mesencephalon - drug effects; Mesencephalon - metabolism; Metals and various inorganic compounds; Mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; Oxidative stress; Oxidative Stress - drug effects; Oxidative Stress - physiology; Rats; Rats, Sprague-Dawley; Toxicology
• ISSN: 0161-813X; 1872-9711
• DOI: 10.1016/S0161-813X(03)00042-1

Dopamine (DA)-induced neurotoxicity is potentiated when cellular metabolism is compromised. Since cyanide is a neurotoxin that produces mitochondrial dysfunction and stimulates intracellular generation of reactive oxygen species (ROS), KCN was used to study DA-induced apoptosis in primary cultured mesencephalon cells. Treatment of neurons with DA (300 μM) for 24 h produced apoptosis as determined by TUNEL staining, DNA fragmentation and increased caspase activity. Pretreatment with KCN (100 μM) 30 min prior to DA increased the number of cells undergoing apoptosis. When added to the cells alone, this concentration of KCN did not induce apoptosis. DA stimulated intracellular generation of ROS, and treatment with KCN enhanced ROS generation. Treatment of cells with glutathione or uric acid (antioxidants/scavengers) attenuated both the increase in ROS generation and the apoptosis, demonstrating that ROS are initiators of the cytotoxicity. Studies on the sequence of events mediating the response showed that DA-induced depolarization of the mitochondrial membrane was dependent on ROS generation and KCN enhanced this action of DA. Following changes in mitochondrial membrane potential, cytochrome c was released from mitochondria, leading to caspase activation and eventually cell death. These results demonstrate that oxidative stress and mitochondrial dysfunction are initiators of DA-induced apoptosis. Subsequent cytochrome c release activates the caspase effector component of apoptosis. Cyanide potentiates the neurotoxicity of DA by enhancing the generation of ROS and impairing mitochondrial function.