Fudge Dylan H, Black Ray, Qin Yan
Department of Biological Sciences, University of Denver, Denver, CO, USA.
Methods Mol Biol. 2018;1867:103-112. doi: 10.1007/978-1-4939-8799-3_8.
In addition to serving as an essential structural component, zinc is also involved in intracellular and intercellular signaling pathways to impact a number of cellular functions. Genetically encoded zinc sensors that are specifically targeted to various subcellular compartments (ER, mitochondria, nucleus, plasma membrane, and vesicles) have been proven to provide accurate and sensitive visualization and quantification of zinc. Here we describe the methods to utilize both ratiometric and intensiometric genetically encoded zinc sensors designed based on zinc fingers for imaging and quantification of cellular free, labile zinc concentrations, [Zn]. This chapter explains in detail how to quantify [Zn] in live cells as well as how to monitor zinc influx in INS-1 cells stimulated with high glucose.
除了作为一种重要的结构成分外，锌还参与细胞内和细胞间信号通路，以影响多种细胞功能。已证实，专门针对各种亚细胞区室（内质网、线粒体、细胞核、质膜和囊泡）的基因编码锌传感器能够准确、灵敏地可视化和定量锌。在这里，我们描述了利用基于锌指设计的比率型和强度型基因编码锌传感器对细胞游离、不稳定锌浓度[Zn]进行成像和定量的方法。本章详细解释了如何定量活细胞中的[Zn]，以及如何监测高糖刺激的INS-1细胞中的锌内流。
