aphidicolin and Amyotrophic-Lateral-Sclerosis

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.

Topics: Amyotrophic Lateral Sclerosis; Animals; Aphidicolin; Cell Differentiation; Cell Survival; Cyclic AMP; Cytochrome c Group; DNA Fragmentation; Enzyme Inhibitors; Humans; Microtubule-Associated Proteins; Motor Neurons; Mutation; Neurites; Neurofilament Proteins; Superoxide Dismutase; Tumor Cells, Cultured