Measuring calcium dynamics in living cells with genetically encodable calcium indicators.
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.
McCombs
Janet E
JE
Department of Chemistry and Biochemistry, University of Colorado, UCB 215, 76 Chemistry, Boulder, CO 80309-0215, USA.
Palmer
Amy E
AE
D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002138:Calibration; D002478:Cells, Cultured; D031541:Fluorescence Resonance Energy Transfer; D007202:Indicators and Reagents; D008164:Luminescent Proteins / Q000378:metabolism; D008958:Models, Molecular; D015388:Organelles / Q000378:metabolism; D044169:Receptors, Calcium-Sensing; D014162:Transfection / Q000379:methods
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D044169:Receptors, Calcium-Sensing; D002118:Calcium