Department of Developmental Biology, School of Medicine, Stanford University, Stanford, California 94305, United States.
Department of Chemical and Systems Biology, School of Medicine, Stanford University, Stanford, California 94305, United States.
ACS Chem Biol. 2022 Aug 19;17(8):2024-2030. doi: 10.1021/acschembio.2c00333. Epub 2022 Jul 15.
cAMP is a ubiquitous second messenger with many functions in diverse organisms. Current cAMP sensors, including Föster resonance energy transfer (FRET)-based and single-wavelength-based sensors, allow for real time visualization of this small molecule in cultured cells and in some cases in vivo. Nonetheless the observation of cAMP in living animals is still difficult, typically requiring specialized microscopes and ex vivo tissue processing. Here we used ligand-dependent protein stabilization to create a new cAMP sensor. This sensor allows specific and sensitive detection of cAMP in living zebrafish embryos, which may enable new understanding of the functions of cAMP in living vertebrates.
cAMP 是一种普遍存在的第二信使,在多种生物中具有多种功能。目前的 cAMP 传感器,包括基于荧光共振能量转移(FRET)和单波长的传感器,允许实时可视化培养细胞中的这种小分子,在某些情况下还可以在体内进行。尽管如此,在活体动物中观察 cAMP 仍然很困难,通常需要专门的显微镜和离体组织处理。在这里,我们使用配体依赖性蛋白稳定化来创建一种新的 cAMP 传感器。该传感器可特异性和灵敏地检测活体斑马鱼胚胎中的 cAMP,这可能使人们对 cAMP 在活体脊椎动物中的功能有新的认识。