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大鼠大脑皮层薄片中钙区室的研究:藜芦定、锂及线粒体解偶联剂的作用

A study of calcium compartments in rat brain cortex thin slices: effects of veratridine, lithium and of a mitochondrial uncoupler.

作者信息

Rubiales de Barioglio S, Orrego F

出版信息

Neurochem Res. 1982 Nov;7(11):1427-35. doi: 10.1007/BF00966071.

DOI:10.1007/BF00966071
PMID:7170060
Abstract

The efflux kinetics of 45Ca from rat brain cortex thin slices previously equilibrated with it, was studied in a superfusion system. Two first order kinetic components of efflux from the tissue were found: k2 = 0.0667 min-1, that was unchanged by lowering the temperature from 37 degrees C to 15 degrees, and k3 = 0.0167 min-1 at 37 degrees C, that was reduced to 0.0897 min-1 at 15 degrees C. This suggests that k2 represents efflux from the extracellular space, and k3 that from the cellular compartment. Addition of the mitochondrial uncoupler carbonyl cyanide, m-chlorophenylhydrazone (CCCP) (10(-5)M) increased the efflux fractional rate constant of 45Ca by 35%, while no change in efflux was induced by 10 mM caffeine. Veratridine (10(-5)M) drastically reduced 45Ca efflux if superfusion was with physiological salt solution (150 mM sodium present), but not if 50 mM lithium replaced an equivalent amount of sodium in the superfusion fluid. This lithium-containing solution did not affect 45Ca efflux in the absence of veratridine. These results indicate that mitochondria accumulate only a minor fraction of intracellular 45Ca; that 45Ca possibly turns over very rapidly in the endoplasmic reticulum, and that most of 45Ca is present in a different, non-mitochondrial, non endoplasmic reticular compartment, the nature of which can be only conjectured.

摘要

在一个灌流系统中,研究了先前已用45Ca平衡过的大鼠大脑皮层薄片中45Ca的流出动力学。发现组织流出有两个一级动力学成分:k2 = 0.0667分钟-1,将温度从37℃降至15℃时该值不变;k3在37℃时为0.0167分钟-1,在15℃时降至0.0897分钟-1。这表明k2代表细胞外空间的流出,而k3代表细胞内区室的流出。加入线粒体解偶联剂羰基氰化物间氯苯腙(CCCP)(10-5M)使45Ca的流出分数速率常数增加了35%,而10 mM咖啡因未引起流出变化。藜芦碱(10-5M)在灌流生理盐溶液(含150 mM钠)时会大幅降低45Ca流出,但当灌流液中50 mM锂替代等量钠时则不会。这种含锂溶液在无藜芦碱时不影响45Ca流出。这些结果表明,线粒体仅积累细胞内45Ca的一小部分;45Ca可能在内质网中周转非常迅速,且大部分45Ca存在于不同的、非线粒体、非内质网的区室中,其性质只能推测。

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本文引用的文献

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THE CALCIUM CONTENT OF ISOLATED CEREBRAL TISSUES AND THEIR STEADY-STATE EXCHANGE OF CALCIUM.离体脑组织的钙含量及其钙的稳态交换
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Sodium-dependent efflux of [3H]GABA from synaptosomes probably related to mitochondrial calcium mobilization.[3H]GABA从突触体的钠依赖性流出可能与线粒体钙动员有关。
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Mg2+ inhibition of Na2+-stimulated Ca2+ release from brain mitochondria.
J Biol Chem. 1980 Jul 25;255(14):6556-8.