Defective neurite outgrowth in aphidicolin/cAMP-induced motor neurons expressing mutant Cu/Zn superoxide dismutase.
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by motor neuron involvement. Mutations in the human Cu/Zn superoxide dismutase (SOD1) gene are found in some cases of familial ALS. Many studies have reported SOD1 mutation-related neurodegeneration. However, whether or not a mutant SOD1 affects neural development has not been demonstrated. We developed motor neuron-neuroblastoma hybrid cells that expressed a mutant (G93A) or the wild type (WT) SOD1. Cells were differentiated by dibutyryl cAMP and aphidicolin. The mutant showed a defect in neurite outgrowth and had decreased viability. Cytochrome c released and nuclear fragmentation were observed. Western blot analysis showed that the amount of neurofilament and microtubule associated proteins-2 (MAP-2) decreased during differentiation. These results suggest that the defect in neurite outgrowth of mutant SOD1 cells is a cytoskeletal defect and is associated with neuronal death.
Lee
Kwang-Woo
KW
Department of Neurology, Neuroscience Center, Seoul National University and Biomedical Research Center, Korean National Institute of Health, Seoul, South Korea.
Kim
Hyun-Jeong
HJ
Sung
Jung-Joon
JJ
Park
Kyung-Seok
KS
Kim
Manho
M
D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology* / Q000235:genetics; D000818:Animals; D016590:Aphidicolin; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics*; D000242:Cyclic AMP; D003574:Cytochrome c Group / Q000187:drug effects / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects / Q000235:genetics; D004791:Enzyme Inhibitors; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000187:drug effects / Q000378:metabolism; D009046:Motor Neurons / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D009154:Mutation / Q000235:genetics*; D016501:Neurites / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D016900:Neurofilament Proteins / Q000187:drug effects / Q000378:metabolism; D013482:Superoxide Dismutase / Q000172:deficiency* / Q000235:genetics; D014407:Tumor Cells, Cultured
D003574:Cytochrome c Group; D004791:Enzyme Inhibitors; D008869:Microtubule-Associated Proteins; D016900:Neurofilament Proteins; D016590:Aphidicolin; D000242:Cyclic AMP; D013482:Superoxide Dismutase