Behera Smrutisanjita, Krebs Melanie, Loro Giovanna, Schumacher Karin, Costa Alex, Kudla Jörg
Institute of Plant Biology and Biotechnology, University of Münster, 48149 Münster, Germany.
Cold Spring Harb Protoc. 2013 Aug 1;2013(8):700-3. doi: 10.1101/pdb.top066183.
Temporally and spatially defined changes in cellular calcium (Ca(2+)) concentration represent stimulus-specific signals and regulate a myriad of biological processes. The development of ratiometric Ca(2+) reporter proteins like Yellow Cameleons (YCs) has greatly advanced our ability to analyze Ca(2+) dynamics in vivo with unprecedented spatial and temporal resolution. In plants, the application of these Ca(2+) reporter proteins has been pioneered for the investigation of Ca(2+) dynamics in guard cells, and recently their use has been extended to other single-cell models like growing pollen tubes and root hairs. However, in plants, the use of YC reporter proteins has largely remained restricted to the investigation of cytoplasmic alterations of Ca(2+) concentrations. Here, we provide an introduction to current methods for imaging Ca(2+) dynamics with increasing sophistication.
细胞钙（Ca(2+)）浓度在时间和空间上的特定变化代表了刺激特异性信号，并调节着无数的生物过程。诸如黄色变色龙（YCs）等比率型Ca(2+)报告蛋白的发展极大地提升了我们在体内以前所未有的时空分辨率分析Ca(2+)动态变化的能力。在植物中，这些Ca(2+)报告蛋白已率先应用于保卫细胞中Ca(2+)动态变化的研究，最近它们的应用已扩展到其他单细胞模型，如生长中的花粉管和根毛。然而，在植物中，YC报告蛋白的使用在很大程度上仍局限于对Ca(2+)浓度细胞质变化的研究。在此，我们介绍目前用于成像Ca(2+)动态变化且日益复杂的方法。
