日本京都大学Masayuki Sakamoto、Tatsushi Yokoyama课题组的最新研究，开发了用于解密体内钙和cAMP群体编码的多色套件。该研究于2024年3月21日发表于国际学术期刊《自然-方法学》杂志。

为了破译体内cAMP的动态变化，研究人员合理地设计了cAMPinG1，它是一种灵敏的基因编码绿色cAMP指示剂，在动态范围和cAMP亲和力方面都优于其前代产品。双光子cAMPinG1成像能在数十秒内，检测到小鼠视觉皮层神经元体节和树突棘的cAMP瞬时变化。

此外，通过使用敏感的红色Ca²⁺指示剂RCaMP3进行多色成像，可以同时测量数百个神经元中Ca²⁺和cAMP的群体模式。研究发现与Ca²⁺相关的cAMP反应代表了特定的信息，如视觉和运动中的方向选择性，以及与GPCR相关的cAMP反应。总之，该研究建立的多色套件将有助于在体外和体内以单细胞分辨率，分析Ca²⁺、GPCR和cAMP信号之间的相互作用。

据了解，cAMP是一种通用的第二信使，受包括Ca²⁺和G蛋白偶联受体(GPCR)在内的各种上游途径的调控。

附：英文原文

Title: A multicolor suite for deciphering population coding of calcium and cAMP in vivo

Author: Yokoyama, Tatsushi, Manita, Satoshi, Uwamori, Hiroyuki, Tajiri, Mio, Imayoshi, Itaru, Yagishita, Sho, Murayama, Masanori, Kitamura, Kazuo, Sakamoto, Masayuki

Issue&Volume: 2024-03-21

Abstract: cAMP is a universal second messenger regulated by various upstream pathways including Ca2+ and G-protein-coupled receptors (GPCRs). To decipher in vivo cAMP dynamics, we rationally designed cAMPinG1, a sensitive genetically encoded green cAMP indicator that outperformed its predecessors in both dynamic range and cAMP affinity. Two-photon cAMPinG1 imaging detected cAMP transients in the somata and dendritic spines of neurons in the mouse visual cortex on the order of tens of seconds. In addition, multicolor imaging with a sensitive red Ca2+ indicator RCaMP3 allowed simultaneous measurement of population patterns in Ca2+ and cAMP in hundreds of neurons. We found Ca2+-related cAMP responses that represented specific information, such as direction selectivity in vision and locomotion, as well as GPCR-related cAMP responses. Overall, our multicolor suite will facilitate analysis of the interaction between the Ca2+, GPCR and cAMP signaling at single-cell resolution both in vitro and in vivo.

DOI: 10.1038/s41592-024-02222-9

Source: https://www.nature.com/articles/s41592-024-02222-9