Measuring calcium dynamics in living cells with genetically encodable calcium indicators.

Measuring calcium dynamics in living cells with genetically encodable calcium indicators.
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DOI:
10.1016/j.ymeth.2008.09.015
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发表时间:
2008-11
期刊:
影响因子:
4.8
通讯作者:
Palmer, Amy E.
Palmer, Amy E.
中科院分区:
生物学3区
文献类型:
--
作者:
McCombs, Janet E.;Palmer, Amy E.

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基因编码钙指示剂 (GECI) 使研究人员能够测量活细胞内特定目标位置的钙动态。这些指标能够剖析钙信号传导过程的空间和时间控制。在这里,我们回顾了 GECI 开发的最新进展,重点介绍了哪些指标最适合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。概述了最近为确保 GECI 受到细胞环境干扰最小化而采取的方法。此外,还讨论了将 GECI 引入哺乳动物细胞、进行钙成像实验和分析数据的程序。由于针对细胞器的指标通常会带来额外的挑战,因此我们强调在这些位置校准 GECI 的策略。
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.
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