Cu/Zn superoxide dismutase (SOD1) mutations associated with familial amyotrophic lateral sclerosis (ALS) affect cellular free radical release in the presence of oxidative stress

Abstract

NO is likely to contribute to motor neuron injury, but this does not fully account for all the cellular toxic effects of mutant SOD1.

Analysis of the Cytosolic Proteome in a Cell Culture Model of Familial Amyotrophic Lateral Sclerosis Reveals Alterations to the Proteasome, Antioxidant Defenses, and Nitric Oxide Synthetic Pathways

“…The nature of the alteration here to the mouse ASL gene product is currently under investigation. By using microelectrode biosensor measurements, we have determined previously that NSC34 cells expressing wild-type hSOD1 exhibit enhanced NO release, whereas those expressing ALS mutant hSOD1 exhibit reduced NO release following cell stress induced by serum withdrawal (29). Both groups of cells expressing wild-type or mutant hSOD1 show decreased superoxide release in the same experimental paradigm.…”

“…Both groups of cells expressing wild-type or mutant hSOD1 show decreased superoxide release in the same experimental paradigm. In addition, the mutant SOD1-expressing NSC34 cells are more sensitive to apoptosis stimulated by NO-releasing compounds (29). The mechanism by which NO release is reduced in the mutant SOD1-expressing NSC34 cells has yet to be determined.…”

“…However, due to the low proportion of motor neurons compared with other cell types in spinal cord, the primary responses of motor neurons to mutant SOD1 toxicity that trigger cell death pathways may go undetected among the overall tissue transcriptional changes responding to motor neuron degeneration. To this end we have exploited a well characterized cell culture model of mutant SOD1-related FALS (27)(28)(29) to enable us to analyze the proteome changes that occur as a direct result of SOD1 toxicity in cells with a motor neuron phenotype.…”

“…Primary alterations within degenerating motor neurons in response to mutant ALS SOD1 may go undetected among the protein changes occurring within the complex mixture of more abundant cell types in the central nervous system. We have addressed this issue by developing a cell culture model of FALS based upon a murine motor neuron-like cell line, NSC34, stably transfected with human FALS associated SOD1 mutants G93A and G37R (27)(28)(29). The NSC34 cells are a hybrid motor neuron ϫ neuroblastoma cell line that exhibits several features of motor neurons including neurofilament expression, the ability to generate action potentials, and induction of twitching in co-cultured muscle cells (30).…”

Vascular Endothelial Growth Factor Attenuates Neurodegenerative Changes in the NSC-34 Motor Neuron Cell Line Induced by Cerebrospinal Fluid of Sporadic Amyotrophic Lateral Sclerosis Patients

“…This cell line is a valuable tool for toxicological investigations on motor neurons as it can be propagated continuously while providing abundant cellular material for biochemical assays [8] . mSOD1-transfected NSC-34 cells have earlier been used to study the mechanisms associated with neurodegeneration in FALS [9] .…”