Travis E R, Wightman R M
Department of Chemistry, University of North Carolina at Chapel Hill 27599-3290, USA.
Annu Rev Biophys Biomol Struct. 1998;27:77-103. doi: 10.1146/annurev.biophys.27.1.77.
Biophysical events involved in late stages of exocytosis occur at highly localized areas of cells on millisecond and submillisecond time scales. Thus, methodologies with high spatio-temporal resolution are required to achieve measurements at individual secretory cells. Much has been learned about the mechanisms and kinetics of vesicular release through analysis with the carbon fiber microelectrode techniques amperometry and cyclic voltammetry. Coupling of these techniques with other methods such as patch-clamp continues to reveal details of the secretion process. It is now clear that extrusion of the vesicular contents is a more complex process than previously believed. Vesicle-cell fusion, revealed by cell capacitance measurements, is temporally dissociated from secretion measured amperometrically. The stability imparted by interaction and association of vesicle contents at rest results in a rate-limiting extrusion process after full fusion. Furthermore, the presence of partial fusion events and the occurrence of nonquantized release have been revealed with electrochemical tools.
胞吐作用后期涉及的生物物理事件发生在细胞的高度局部化区域,时间尺度为毫秒和亚毫秒级。因此,需要具有高时空分辨率的方法来在单个分泌细胞上进行测量。通过使用碳纤维微电极技术安培法和循环伏安法进行分析,人们对囊泡释放的机制和动力学有了很多了解。这些技术与其他方法(如膜片钳)的结合不断揭示分泌过程的细节。现在很清楚,囊泡内容物的挤出是一个比以前认为的更复杂的过程。通过细胞电容测量揭示的囊泡 - 细胞融合在时间上与通过安培法测量的分泌解离。静止时囊泡内容物的相互作用和缔合赋予的稳定性导致完全融合后的限速挤出过程。此外,电化学工具已经揭示了部分融合事件的存在和非量化释放的发生。