An Expanded Palette of Genetically Encoded Ca2+ Indicators
An Expanded Palette of Genetically Encoded Ca2+ Indicators
复制标题
DOI：
10.1126/science.1208592
复制
发表时间：
2011-09-30
期刊：
影响因子：
56.9
通讯作者：
Campbell, Robert E.
中科院分区：
文献类型：
--
作者：
Zhao, Yongxin;Araki, Satoko;Campbell, Robert E.
工程化荧光蛋白（FP）嵌合体，调节其荧光响应钙离子（Ca 2+）浓度的变化是可视化细胞内信号传导活性的有力工具。然而，尽管已有十年的可用性，但单一基于FP的Ca 2+指示剂的调色板仍然限于单一的绿色色调。我们通过开发蓝色、改进的绿色和红色强度指示器以及具有11，000%比率变化的排放比率指示器来扩展该调色板。这一系列能够改善神经元和转基因秀丽隐杆线虫的单色Ca 2+成像。在HeLa细胞中，Ca 2+在三个亚细胞区室中成像，并且结合基于青色FP-黄色FP的指示剂，Ca 2+和腺苷5 '-三磷酸同时成像。这个调色板的指标描绘了一个丰富多彩的新时代的Ca 2+成像的方式。
Engineered fluorescent protein (FP) chimeras that modulate their fluorescence in response to changes in calcium ion (Ca2+) concentration are powerful tools for visualizing intracellular signaling activity. However, despite a decade of availability, the palette of single FP-based Ca2+ indicators has remained limited to a single green hue. We have expanded this palette by developing blue, improved green, and red intensiometric indicators, as well as an emission ratiometric indicator with an 11,000% ratio change. This series enables improved single-color Ca2+ imaging in neurons and transgenic Caenorhabditis elegans. In HeLa cells, Ca2+ was imaged in three subcellular compartments, and, in conjunction with a cyan FP-yellow FP-based indicator, Ca2+ and adenosine 5'-triphosphate were simultaneously imaged. This palette of indicators paints the way to a colorful new era of Ca2+ imaging.