Prole David L, López-Sanjurjo Cristina I, Tovey Stephen C, Taylor Colin W
Department of Pharmacology, University of Cambridge, Cambridge, UK.
Methods Cell Biol. 2015;126:237-59. doi: 10.1016/bs.mcb.2014.10.024. Epub 2015 Jan 16.
The endoplasmic reticulum (ER) is both the major source of intracellular Ca(2+) for cell signaling and the organelle that forms the most extensive contacts with the plasma membrane and other organelles. Lysosomes fulfill important roles in degrading cellular materials and in cholesterol handling, but they also contribute to Ca(2+) signaling by both releasing and sequestering Ca(2+). Interactions between ER and other Ca(2+)-transporting membranes, notably mitochondria and the plasma membrane, often occur at sites where the two membranes are closely apposed, allowing local Ca(2+) signaling between them. These interactions are often facilitated by scaffold proteins. Recent evidence suggests similar local interactions between ER and lysosomes. We describe simple fluorescence-based methods that allow the interplay between Ca(2+) signals, the ER, and lysosomes to be examined.
内质网（ER）既是细胞信号传导中细胞内Ca(2+)的主要来源，也是与质膜和其他细胞器形成最广泛接触的细胞器。溶酶体在降解细胞物质和胆固醇处理中发挥重要作用，但它们也通过释放和螯合Ca(2+)参与Ca(2+)信号传导。内质网与其他Ca(2+)转运膜（特别是线粒体和质膜）之间的相互作用通常发生在两个膜紧密相邻的部位，从而允许它们之间进行局部Ca(2+)信号传导。这些相互作用通常由支架蛋白促进。最近的证据表明内质网与溶酶体之间存在类似的局部相互作用。我们描述了简单的基于荧光的方法，可用于检测Ca(2+)信号、内质网和溶酶体之间的相互作用。
