使用基因编码钙指示剂测量活细胞中的钙动力学。
Measuring calcium dynamics in living cells with genetically encodable calcium indicators.
作者信息
McCombs Janet E, Palmer Amy E
机构信息
Department of Chemistry and Biochemistry, University of Colorado, UCB 215, 76 Chemistry, Boulder, CO 80309-0215, USA.
出版信息
Methods. 2008 Nov;46(3):152-9. doi: 10.1016/j.ymeth.2008.09.015. Epub 2008 Oct 9.
Abstract
Genetically encoded calcium indicators (GECIs) allow researchers to measure calcium dynamics in specific targeted locations within living cells. Such indicators enable dissection of the spatial and temporal control of calcium signaling processes. Here we review recent progress in the development of GECIs, highlighting which indicators are most appropriate for measuring calcium in specific organelles and localized domains in mammalian tissue culture cells. An overview of recent approaches that have been undertaken to ensure that the GECIs are minimally perturbed by the cellular environment is provided. Additionally, the procedures for introducing GECIs into mammalian cells, conducting calcium imaging experiments, and analyzing data are discussed. Because organelle-targeted indicators often pose an additional challenge, we underscore strategies for calibrating GECIs in these locations.
摘要
基因编码钙指示剂（GECIs）使研究人员能够测量活细胞内特定靶向位置的钙动态变化。这类指示剂有助于剖析钙信号传导过程的时空控制机制。在此，我们综述了GECIs开发方面的最新进展，重点介绍了哪些指示剂最适合用于测量哺乳动物组织培养细胞中特定细胞器和局部区域的钙含量。本文还概述了为确保GECIs受细胞环境干扰最小而采取的最新方法。此外，还讨论了将GECIs导入哺乳动物细胞、进行钙成像实验及分析数据的步骤。由于细胞器靶向指示剂往往带来额外挑战，我们强调了在这些位置校准GECIs的策略。
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