Integrating Optical and Electrochemical Approaches to Assess the Actions of Dynamin at the Fusion Pore.
Integrating Optical and Electrochemical Approaches to Assess the Actions of Dynamin at the Fusion Pore.
复制标题
综合光学和电化学方法来评估 Dynamin 在融合孔中的作用。
DOI：
10.1007/978-1-0716-0676-6_12
复制
发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Anantharam,Arun
中科院分区：
文献类型：
--
作者：
Smith,KatherineA;Prantzalos,EmilyR;Anantharam,Arun
在目前可用于监测肾上腺嗜铬细胞中致密核心颗粒胞吐的技术中，有两种被证明特别有用：碳纤维安培法和全内反射荧光(TIRF)显微镜。安培法能够以毫秒的时间分辨率检测从融合孔中逃脱的可氧化儿茶酚胺。TIRF显微镜及其变种的偏振TIRF(PTIRF)显微镜提供了有关融合孔在时间上的后期特征的信息。结合使用，安培测量和TIRF显微镜允许研究人员跟踪融合孔从形成到扩张或重新闭合的命运。融合孔的性质，包括它们的结构和动力学，已经被多个被Dynamin GTPase(Dyn1)修饰的基团所展示。在本章中，我们描述了安培法和TIRF显微镜如何使我们深入了解动力蛋白对牛肾上腺嗜铬细胞原代培养的胞吐作用的影响。
Of the techniques currently available to monitor dense core granule exocytosis in adrenal chromaffin cells, two have proven particularly useful: carbon-fiber amperometry and total internal reflection fluorescence (TIRF) microscopy. Amperometry enables the detection of oxidizable catecholamines escaping a fusion pore with millisecond time resolution. TIRF microscopy, and its variant polarized-TIRF (pTIRF) microscopy, provides information on the characteristics of fusion pores at temporally later stages. Used in conjunction, amperometry and TIRF microscopy allow an investigator to follow the fate of a fusion pore from its formation to expansion or reclosure. The properties of fusion pores, including their structure and dynamics, have been shown by multiple groups to be modified by the dynamin GTPase (Dyn1). In this chapter, we describe how amperometry and TIRF microscopy enable insights into dynamin-dependent effects on exocytosis in primary cultures of bovine adrenal chromaffin cells.