Wightman R M, Schroeder T J, Finnegan J M, Ciolkowski E L, Pihel K
Department of Chemistry, University of North Carolina at Chapel Hill 27599-3290.
Biophys J. 1995 Jan;68(1):383-90. doi: 10.1016/S0006-3495(95)80199-2.
The time course of extrusion of the vesicular contents during exocytosis has been examined at adrenal medullary cells with carbon-fiber microelectrodes. Two electrochemical techniques were used: cyclic voltammetry and amperometry. Spikes obtained by amperometry had a faster time course than those measured by cyclic voltammetry, consistent with the different concentration profiles established by each technique. However, the experimental data obtained with both techniques were temporally broadened with respect to dispersion of an instantaneous point source by diffusion. Measurements with the electrode firmly pressed against the cell surface established that the temporal broadening is a result of a rate-limiting kinetic step associated with extrusion of the vesicular contents at the cell surface. The data do not support a rate-limiting process due to restricted efflux from a small pore. When combined with previous results, the data suggest that the rate-limiting step for chemical secretion from adrenal medullary cells during exocytosis is the dissociation of catecholamines from the vesicular matrix at the surface of the cell.
利用碳纤维微电极在肾上腺髓质细胞上研究了胞吐作用期间囊泡内容物挤出的时间进程。使用了两种电化学技术:循环伏安法和安培法。通过安培法获得的尖峰比通过循环伏安法测量的尖峰具有更快的时间进程,这与每种技术建立的不同浓度分布一致。然而,两种技术获得的实验数据相对于瞬时点源通过扩散的分散在时间上都变宽了。将电极紧紧压在细胞表面进行测量表明,时间变宽是与细胞表面囊泡内容物挤出相关的限速动力学步骤的结果。数据不支持由于从小孔的流出受限而导致的限速过程。与先前的结果相结合,数据表明肾上腺髓质细胞在胞吐作用期间化学分泌的限速步骤是细胞表面儿茶酚胺从囊泡基质的解离。