Jankowski J A, Finnegan J M, Wightman R M
Department of Chemistry, University of North Carolina at Chapel Hill 27599-3290.
J Neurochem. 1994 Nov;63(5):1739-47. doi: 10.1046/j.1471-4159.1994.63051739.x.
The temporal resolution of carbon-fiber microelectrodes has been exploited to examine the plasticity of quantal secretory events at individual adrenal medullary cells. The size of individual quantal events, monitored by amperometric oxidation of released catecholamines, was found to be dependent on the extracellular ionic composition, the secretagogue, and the order of depolarization delivery. Release was observed with either exposure to 60 mM K+ in the presence of Ca2+ or exposure to 3 mM Ba2+ in solutions of different pH, with and without external Ca2+. Ba2+ was demonstrated to induce Ca(2+)-independent exocytotic release for an extended period of time (> 4 min) relative to release induced by K+ (approximately 30 s), which is Ca2+ dependent. In all cases, simultaneous changes of intracellular divalent cations, monitored by fura-2 fluorescence, accompanied quantal release and had a similar time course. Exocytosis caused by Ba2+ in Ca(2+)-free medium had a larger mean spike area at pH 8.2 than at pH 7.4. When Ba(2+)-induced spikes measured at pH 7.4 were compared, the spikes in Ca(2+)-free medium were found to be broader and shorter but had the same area. Release induced by K+ after exposure to Ba2+ was comprised of larger quantal events when compared with preceding K+ stimulations. Finally, spikes obtained with Ba2+ exposure at an extracellular pH of 5.5 had a different shape than those obtained in more basic solutions. These changes in spike size and shape are consistent with the interactions between catecholamines and other intravesicular components.
利用碳纤维微电极的时间分辨率来研究单个肾上腺髓质细胞中量子分泌事件的可塑性。通过对释放的儿茶酚胺进行安培氧化监测的单个量子事件的大小,发现其取决于细胞外离子组成、促分泌剂以及去极化传递的顺序。在存在Ca2+的情况下暴露于60 mM K+或在不同pH值的溶液中暴露于3 mM Ba2+(有无外部Ca2+)时均观察到释放。相对于K+诱导的释放(约30秒,依赖于Ca2+),Ba2+被证明可诱导长时间(>4分钟)的不依赖Ca(2+)的胞吐释放。在所有情况下,通过fura-2荧光监测的细胞内二价阳离子的同时变化伴随着量子释放,并且具有相似的时间进程。在无Ca(2+)的培养基中,Ba2+引起的胞吐作用在pH 8.2时的平均尖峰面积比在pH 7.4时更大。当比较在pH 7.4下测量的Ba(2+)诱导的尖峰时,发现无Ca(2+)培养基中的尖峰更宽且更短,但面积相同。与之前的K+刺激相比,暴露于Ba2+后K+诱导的释放由更大的量子事件组成。最后,在细胞外pH为5.5时暴露于Ba2+获得的尖峰形状与在更碱性溶液中获得的尖峰形状不同。尖峰大小和形状的这些变化与儿茶酚胺和其他囊泡内成分之间的相互作用一致。