Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro, Tokyo, 153-8902, Japan.
Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan.
Sci Rep. 2017 Aug 4;7(1):7351. doi: 10.1038/s41598-017-07820-6.
cAMP is a common second messenger that is involved in various physiological processes. To expand the colour palette of available cAMP indicators, we developed a red cAMP indicator named "Pink Flamindo" (Pink Fluorescent cAMP indicator). The fluorescence intensity of Pink Flamindo increases 4.2-fold in the presence of a saturating dose of cAMP, with excitation and emission peaks at 567 nm and 590 nm, respectively. Live-cell imaging revealed that Pink Flamindo is effective for monitoring the spatio-temporal dynamics of intracellular cAMP generated by photoactivated adenylyl cyclase in response to blue light, and in dual-colour imaging studies using a green Ca indicator (G-GECO). Furthermore, we successfully monitored the elevation of cAMP levels in vivo in cerebral cortical astrocytes by two-photon imaging. We propose that Pink Flamindo will facilitate future in vivo, optogenetic studies of cell signalling and cAMP dynamics.
cAMP 是一种常见的第二信使，参与各种生理过程。为了扩展可用 cAMP 指示剂的颜色范围，我们开发了一种红色 cAMP 指示剂，命名为“Pink Flamindo”（粉色荧光 cAMP 指示剂）。在存在饱和剂量的 cAMP 时，Pink Flamindo 的荧光强度增加 4.2 倍，激发和发射峰分别在 567nm 和 590nm 处。活细胞成像显示，Pink Flamindo 可有效监测光激活腺苷酸环化酶响应蓝光产生的细胞内 cAMP 的时空动力学，以及与绿色 Ca 指示剂（G-GECO）的双色成像研究。此外，我们还成功地通过双光子成像监测到体内大脑皮质星形胶质细胞中 cAMP 水平的升高。我们提出，Pink Flamindo 将有助于未来的细胞信号转导和 cAMP 动力学的体内光遗传学研究。
