Kawata Seiko, Yamamoto Yuto, Saitoh Naoto
Department of Neurophysiology, Graduate School of Life and Medical Sciences, Doshisha University, Kyoto 610-0394, Japan.
Data Brief. 2022 Jul 4;43:108441. doi: 10.1016/j.dib.2022.108441. eCollection 2022 Aug.
Cyclic adenosine monophosphate (cyclic AMP) is a second messenger, which is involved in the regulation of various cellular processes, including neuronal firing rate, synaptic plasticity, axon formation and axon elongation in brain. Although the main molecules in the cAMP-mediated signaling pathway are well studied, the spatio-temporal dynamics of the cAMP remain to be elucidated. Live imaging is an informative tool to investigate the cell signaling dynamics. It allows continuous monitoring of a specific cell over a period of time. Thus, optical probes for cAMP are important tools for studying the dynamics of cAMP signaling. Multiple genetically encoded cAMP probes are available [1], [2], including Förster resonance energy transfer (FRET) based or circular permutated fluorescent protein (cpFP) based probes. cpFP-based probes have an advantage of easier handling than FRET-based probes caused by monomeric detection and smaller molecular size. However, there is no cAMP probe compatible with violet light excitation. Therefore, we fused violet light excitable cpGFP to cyclic nucleotide binding domain (CBD) in cAMP receptor protein. This construct successfully responded to cAMP concentration changes. We show here the spectra data and live-cell imaging data of the violet light excitable cAMP probe which can be used for multi-signal fluorescence imaging.
环磷酸腺苷（cAMP）是一种第二信使，参与多种细胞过程的调节，包括大脑中的神经元放电频率、突触可塑性、轴突形成和轴突伸长。尽管cAMP介导的信号通路中的主要分子已得到充分研究，但cAMP的时空动态仍有待阐明。实时成像为研究细胞信号动态提供了一种信息丰富的工具。它可以在一段时间内对特定细胞进行连续监测。因此，用于cAMP的光学探针是研究cAMP信号动态的重要工具。有多种基因编码的cAMP探针可供使用[1,2]，包括基于荧光共振能量转移（FRET）或基于环形排列荧光蛋白（cpFP）的探针。基于cpFP的探针具有比基于FRET的探针更易于操作的优点，这是由于其单体检测和更小的分子尺寸。然而，目前尚无与紫光激发兼容的cAMP探针。因此，我们将紫光可激发的cpGFP与cAMP受体蛋白中的环核苷酸结合结构域（CBD）融合。该构建体成功地对cAMP浓度变化做出了反应。我们在此展示了可用于多信号荧光成像的紫光可激发cAMP探针的光谱数据和活细胞成像数据。
