INTRACELLULAR CALCIUM CONCENTRATIONS DURING METABOLIC INHIBITION IN THE MOTONEURON CELL-LINE NSC-19

被引:12
作者
HASHAM, MI
NAUMANN, D
KIM, SU
CASHMAN, NR
QUAMME, GA
KRIEGER, C
机构
[1] UNIV BRITISH COLUMBIA HOSP,DEPT MED,DIV NEUROL,VANCOUVER V6T 2B5,BC,CANADA
[2] MCGILL UNIV,MONTREAL NEUROL INST,DEPT NEUROL & NEUROSURG,MONTREAL,PQ,CANADA
关键词
CALCIUM; ATP; AMOBARBITAL; CARBONYLCYANIDE M-CHLOROPHENYLHYDRAZONE; MITOCHONDRIA; MOTONEURONS; CELL DEATH;
D O I
10.1139/y94-104
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
Changes in the concentrations of intracellular free calcium ([Ca2+](i)) and adenine nucleotides were determined in response to metabolic inhibitors in the motoneuron cell line NSC-19. The NADH dehydrogenase inhibitor amobarbital (Amytal) and the mitochondrial uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) were used to alter energy metabolism. Exposure of cells to 5 mM Amytal did not significantly change ATP concentrations but produced transient elevations of [Ca2+](i) of approximately 80 nM, which were reduced by 32% when cells were studied in Ca2+-free solutions. CCCP (10 mu M) caused a transient reduction in ATP concentration of 33%. CCCP also produced sustained elevations of [Ca2+](i) of about 280 nM, which were reduced by 47% when in Ca2+-free solutions. In spite of the sustained elevation of [Ca2+](i) induced by CCCP, NSC-19 showed no reduction in cell viability after 48 h compared with controls. Ruthenium red, a blocker of Ca2+ uptake by mitochondria, had little effect on the CCCP-induced [Ca2+](i) increment. KCl or glutamate did not produce significant changes in [Ca2+](i), indicating that these cells do not possess significant numbers of voltage-dependent Ca2+ channels or excitatory amino acid receptor-gated channels. [Ca2+](i) values in these cells were modified by changes in extracellular Ca2+ concentrations. In Ca2+-containing solutions, inhibition of Na+/Ca2+ exchange by amiloride and bepridil led to increased [Ca2+](i), as did blockade of Ca2+ ATPase by vanadate, suggesting that membrane transporters are important in Ca2+ efflux in NSC-19. The present studies indicate that exposure of NSC-19 cells to Amytal and CCCP produces Ca2+ increments by release from internal stores, as well as by transmembrane influx. These results demonstrate that small increments in [Ca2+](i) can be produced by metabolic inhibitors or other compounds and that such changes are not associated with immediate cell death. Changes in [Ca2+](i) could potentially result in abnormal cell function secondary to altered action of Ca2+-dependent enzymes.
引用
收藏
页码:728 / 737
页数:10
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