Okada M, Mine K, Fujiwara M
Department of Physiology and Pharmacology, Faculty of Pharmaceutical Sciences, Fukuoka University, Japan.
J Pharmacol Exp Ther. 1990 Mar;252(3):1283-8.
The involvement of intrasynaptosomal-free Ca++ concentration [( Ca++]i) in Na(+)-dependent release of endogenous dopamine and noradrenaline from rat brain synaptosomes was studied. The release of endogenous dopamine and noradrenaline from rat whole brain synaptosomes were measured using high-performance liquid chromatography with electrochemical detector. The change of [Ca++]i was measured fluorometrically using a Ca++ indicator, Quin-2. Whether extracellular Ca++ was present or not, 30 microM veratridine, a Na(+)-ionophore, increased the release of endogenous dopamine and noradrenaline. In the presence of 1.25 mM Ca++, 30 microM veratridine increased [Ca++]i. In contrast, in the absence of extracellular Ca++, veratridine did not affect [Ca++]i. Ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA) increased the release of dopamine and noradrenaline in Ca-Mg-free medium. This stimulatory effect of EGTA seemed to be the result of an increase in the influx of Na+ through Ca++ channels in the absence of divalent cation. In Ca-Mg-free medium, EGTA caused a slight decrease in [Ca++]i. The EGTA-stimulated release of dopamine and noradrenaline was blocked by La which also significantly blocked the decrease in [Ca++]i observed after the addition of EGTA. These results suggest that the Na(+)-dependent release of dopamine and noradrenaline may not depend on a change in [Ca++]i. Veratridine (30 microM)-induced release of dopamine and noradrenaline were detected simultaneously. However, the time needed to induce the maximal stimulatory effect of veratridine on the release of dopamine was apparently shorter than that of noradrenaline. This delay might suggest that the Na(+)-dependent release process of dopamine is not similar to that of noradrenaline.
研究了突触小体游离钙离子浓度[Ca++]i在大鼠脑突触小体中钠依赖性内源性多巴胺和去甲肾上腺素释放过程中的作用。采用高效液相色谱-电化学检测法测定大鼠全脑突触小体中内源性多巴胺和去甲肾上腺素的释放。使用钙离子指示剂喹啉-2通过荧光法测定[Ca++]i的变化。无论细胞外是否存在钙离子,30微摩尔藜芦碱(一种钠离子载体)均可增加内源性多巴胺和去甲肾上腺素的释放。在存在1.25毫摩尔钙离子的情况下,30微摩尔藜芦碱可增加[Ca++]i。相反,在无细胞外钙离子的情况下,藜芦碱不影响[Ca++]i。乙二醇双(β-氨基乙醚)-N,N'-四乙酸(EGTA)在无钙镁培养基中增加了多巴胺和去甲肾上腺素的释放。EGTA的这种刺激作用似乎是在没有二价阳离子的情况下,通过钙离子通道增加钠离子内流的结果。在无钙镁培养基中,EGTA导致[Ca++]i略有下降。EGTA刺激的多巴胺和去甲肾上腺素释放被镧阻断,镧也显著阻断了加入EGTA后观察到的[Ca++]i的下降。这些结果表明,多巴胺和去甲肾上腺素的钠依赖性释放可能不依赖于[Ca++]i的变化。同时检测到藜芦碱(30微摩尔)诱导的多巴胺和去甲肾上腺素释放。然而,藜芦碱诱导多巴胺释放达到最大刺激作用所需的时间明显短于去甲肾上腺素。这种延迟可能表明多巴胺的钠依赖性释放过程与去甲肾上腺素不同。