Zerby S E, Ewing A G
Department of Chemistry, Penn State University, University Park 16802, USA.
J Neurochem. 1996 Feb;66(2):651-7. doi: 10.1046/j.1471-4159.1996.66020651.x.
To compare the time course of different mechanisms of chemically stimulated release, amperometric detection of dopamine was carried out at single PC12 cells. The rapid response of carbon fiber microelectrodes allowed the detection of single exocytotic events, thus providing time-resolved information about the dynamics of stimulated release, in particular the latency between the stimulation of a cell and the secretion of catecholamines. On rapid depolarization of the cell membrane caused by application of 105 mM K+, almost immediate (6 +/- 1 s) release of dopamine was observed. Stimulation with 1 mM nicotine, involving the stimulant binding to a ligand-gated ion channel, resulted in a short (37 +/- 5 s) delay between stimulation and secretion. Application of 1 mM muscarine to the cells caused a long (103 +/- 11 s) latency before exocytosis was detected. A biphasic response that appeared to be similar to a combination of nicotine- and muscarine-stimulated release was observed when cells were stimulated with 10 mM acetylcholine. Thus, it appears that the dynamics of stimulated release at single PC12 cells is significantly affected by the mechanism leading to exocytosis.
为了比较化学刺激释放的不同机制的时间进程,在单个PC12细胞上进行了多巴胺的安培检测。碳纤维微电极的快速响应使得能够检测单个胞吐事件,从而提供有关刺激释放动力学的时间分辨信息,特别是细胞刺激与儿茶酚胺分泌之间的延迟。在施加105 mM K+导致细胞膜快速去极化时,观察到多巴胺几乎立即(6±1秒)释放。用1 mM尼古丁刺激,涉及兴奋剂与配体门控离子通道结合,导致刺激与分泌之间有短暂(37±5秒)延迟。向细胞施加1 mM毒蕈碱导致在检测到胞吐作用之前有较长(103±11秒)的潜伏期。当用10 mM乙酰胆碱刺激细胞时,观察到一种双相反应,似乎类似于尼古丁和毒蕈碱刺激释放的组合。因此,看来单个PC12细胞上刺激释放的动力学受到导致胞吐作用的机制的显著影响。