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标题：Highlighted Ca2+ imaging with a genetically encoded ‘caged’ indicator
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作者：Tomoki Matsuda ; Kazuki Horikawa ; Kenta Saito 等
期刊名称：Scientific Reports
电子版ISSN：2045-2322
出版年度：2013
卷号：3
期号：1
DOI：10.1038/srep01398
语种：English
出版社：Springer Nature
摘要：Genetically encoded fluorescent indicators for bioimaging are powerful tools for visualizing biological phenomena in specified cell types or cellular compartments. However, available gene promoters or localization sequences are not applicable for visualizing all expression events. Furthermore, a visualization technique focusing on single cells or cellular compartments is required for characterizing specific cellular properties including individuality of cells in the cell population. To address these limitations, we developed a genetically encoded caged Ca²⁺ indicator for which expression timing and location could be controlled. This indicator, PA-TNXL, comprises a Ca²⁺-binding protein and troponin between a photoactivatable FRET donor (PA-GFP) and a FRET quencher (dim variant of YFP). Ultraviolet irradiation activates the FRET Ca²⁺ indicator. Using this indicator, we successfully imaged Ca²⁺ dynamics in a given set of HeLa cells and cultured hippocampal neurons. This technology can be applied for developing other photoactivatable indicators, thereby opening a new area of biological research.