使用荧光指示剂喹啉-2测量突触小体内的游离钙。
Measurement of intrasynaptosomal free calcium by using the fluorescent indicator quin-2.
作者信息
Ashley R H, Brammer M J, Marchbanks R
出版信息
Biochem J. 1984 Apr 1;219(1):149-58. doi: 10.1042/bj2190149.
Abstract
The recently synthesized calcium indicator quin -2 was incorporated into synaptosomes from guinea-pig cerebral cortex following uptake and internal hydrolysis of quin -2 tetra-acetoxymethyl ester. Incubation in physiological media containing 1 mM- or 2 mM-CaCl2 led to equilibrium cytosolic ionized calcium concentrations of 85 +/- 10 nM and 205 +/- 5 nM respectively (mean +/- S.E.M. from eight and eighteen preparations respectively). Cytosolic Ca2+ was elevated following increases in external Ca2+ concentration, plasma membrane depolarization, mitochondrial inhibition, calcium ionophore addition or replacement of external sodium by lithium. Preliminary experiments were performed to assess changes in cytosolic Ca2+ accompanying the release of the neurotransmitter acetylcholine.
摘要
最近合成的钙指示剂喹啉-2在喹啉-2四乙酰氧基甲酯被摄取并进行内部水解后，被整合到豚鼠大脑皮层的突触体中。在含有1 mM或2 mM氯化钙的生理介质中孵育，分别导致胞质游离钙浓度达到85±10 nM和205±5 nM（分别来自8个和18个样本的平均值±标准误）。随着细胞外钙浓度升高、质膜去极化、线粒体抑制、添加钙离子载体或用锂替代细胞外钠，胞质Ca2+浓度会升高。进行了初步实验以评估伴随神经递质乙酰胆碱释放时胞质Ca2+的变化。
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