Greotti Elisa, De Stefani Diego
Neuroscience Institute, National Research Council (CNR), Padova, Italy; Department of Biomedical Sciences, University of Padova, Padova, Italy.
Department of Biomedical Sciences, University of Padova, Padova, Italy.
Methods Cell Biol. 2020;155:337-368. doi: 10.1016/bs.mcb.2019.11.001. Epub 2019 Nov 22.
Calcium (Ca) is a universal intracellular messenger capable of governing a plethora of different biological functions. Its versatility is guaranteed on the one hand by a cell type-specific Ca signaling toolkit. On the other hand, the fine compartmentalization of changes in Ca concentration ([Ca]) into specific subcellular domains adds a level of complexity, thus generating a variety of signals that can be differentially decoded into specific cellular events. In this context, mitochondrial Ca dynamics plays a central role, by regulating both specific organelle functions (e.g., regulation of substrate oxidation, release of caspase cofactors) and global cellular events (e.g., shaping of cytoplasmic Ca waves). Here we describe a general method for the detection of intramitochondrial [Ca] using bioluminescent and fluorescent genetically-encoded Ca indicators (GECIs). We will discuss the characteristics of different GECIs, as well as their strengths, limitations and applications.
钙（Ca）是一种通用的细胞内信使，能够调控众多不同的生物学功能。一方面，细胞类型特异性的钙信号传导工具包确保了其多功能性。另一方面，钙浓度（[Ca]）变化在特定亚细胞区域的精细区室化增加了一层复杂性，从而产生了多种信号，这些信号可以被差异解码为特定的细胞事件。在这种情况下，线粒体钙动力学通过调节特定的细胞器功能（如底物氧化的调节、半胱天冬酶辅因子的释放）和整体细胞事件（如细胞质钙波的形成）发挥核心作用。在这里，我们描述了一种使用生物发光和荧光基因编码钙指示剂（GECIs）检测线粒体内[Ca]的通用方法。我们将讨论不同GECIs的特性，以及它们的优点、局限性和应用。
