Amyotrophic Lateral Sclerosis is a debhilitating and progressive neurodegenerative disease whereby the root causes remain elusive. Oxidative stress has been highlighted as a key player in the pathology of the disease over the past decade, and more recently the development of nitric oxide causing the nitro-tyrosination of proteins in aging and disease has come to light. Another associated factor with the disease is proffessional sportsmen, whom are dispropotionally affected by the disease.Thus, we believe that oxidative stress in combination with mechanical stress exerted on the neuron through intense activity, aided by the presence of androgens, could predispose an individual to developing the disease. We used neuroblasoma cell lines NSC-34 cells and SH-Sy5Y and primary mouse motor neuron cultures to model these effects. We found that hydrogen peroxide produced a significant decrease in cell viability of both cell lines, measured by MTT and that a calcium response could be evoked when applying mechanical stress to both NSC-34 and neuronal cells. We have developed a method for the high purity extraction of motor neurons and a immunocytochomical staining protocol that can be applied to study these pathways and their effect in this debhilitating disease.
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