J Korean Neurol Assoc > Volume 21(5); 2003 > Article

Journal of the Korean Neurological Association 2003;21(5): 521-526.

"변이된 Cu/Zn superoxide dismutase를 발현하는 운동신경세포주에 칼슘조절제가 미치는 영향"

김성훈 , 김현정 김만호 이광우

강원대학교 의과대학 신경과학교실, 서울대학교 의과대학 신경과학교실

"The Effect of Calcium Modulators on Motoneuron Cells Which Express Mutated Cu / Zn Superoxide Dismutase"

Sung Hun Kim

"Department of Neurology, College of Medicine, Kangwon National University* Department of Neurology, College of Medicine, Seoul National University"

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. R e s u l t s: 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. C o n c l u s I o n s: 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.Key Words : Amyotrophic lateral sclerosis, Superoxide dismutase, Calcium, Cell death, Apoptosis, Caspase"