The Effect of Calcium Modulators on Motoneuron Cells Which Express Mutated Cu / Zn Superoxide Dismutase.
- Author:
Sung Hun KIM
1
;
Hyun Jeong KIM
;
Manho KIM
;
Kwang Woo LEE
Author Information
1. Department of Neurology, College of Medicine, Kangwon National University, Korea.
- Publication Type:Original Article
- Keywords:
Amyotrophic lateral sclerosis;
Superoxide dismutase;
Calcium;
Cell death;
Apoptosis;
Caspase
- MeSH:
Amyotrophic Lateral Sclerosis;
Apoptosis;
Calcium*;
Caspase 3;
Cell Death;
Cell Membrane;
Cell Survival;
Enzyme Assays;
Humans;
Motor Neurons;
Neurons;
Superoxide Dismutase*;
Superoxides*;
Thapsigargin
- From:Journal of the Korean Neurological Association
2003;21(5):521-526
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Intracellular calcium accumulation and dys-regulation is known as one of the mechanism in motor neuronal degeneration with mutations in Cu/Zn superoxide dismutase (SOD-1). METHODS: To investigate the calcium modulators and mutated SOD-1 induced neuronal death, we tested motoneuron-neuroblastoma hybrid (VSC 4.1) cells constitutively expressing human SOD-1 gene with mutations (A4V, G93A) or wild type. Calcium mobilizer through cell membrane (calcium ionophore; A23187) or endogenous calcium releaser (ryanodine, thapsigargin, cyclic ADP-ribose) was treated and cell viabilities were determined by using a 3-(4, 5-dimethylthiazole-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Apoptotic cell death was monitored by caspase enzyme assay and the Hoescht staining to check the evidence of nuclear fragmentation. RESULTS: Among agents, calcium ionophore and tharpsigargin reduced the level of survival in cells expressing mutant SOD-1. In mutant cells, caspase 3 activity was elevated and showed nuclear fragmentation. This phenomenon was blocked by caspase inhibitor. CONCLUSIONS: Our data suggest that motor neuron degeneration in familial amyotrophic lateral sclerosis with SOD-1 mutation may be mediated by calcium dys-regulation, particularly by exogenous calcium influx and this process induces caspase 3 activity which results in motor neuron death.
