小柯机器人

美国俄勒冈健康与科学大学Haining Zhong和Tianyi Mao共同合作近期取得重要工作进展，他们研究开发出用于体内cAMP群体和亚细胞成像的灵敏遗传编码传感器。相关论文2022年10月27日在线发表于《自然—方法学》杂志上。

研究人员筛选了现有的基因编码的cAMP传感器，并进一步开发了表现最好的，得出了三个改进的变体，称为cAMPFIREs。与亲本传感器相比，这些传感器对cAMP和胞质分布的敏感性提高了10倍。cAMPFIREs与比率测定法和荧光寿命成像法兼容，并能在生理相关的纳摩尔浓度下检测由去甲肾上腺素引起的cAMP动态变化。对清醒小鼠的cAMPFIREs成像显示，与清醒相关的皮层神经元中cAMP的紧张性水平受到阿片类药物的调节，并在亚细胞区间进行差异调节。

此外，强制运动诱发神经元特异性的双向cAMP动力学。cAMPFIREs也在果蝇中发挥作用。总之，cAMPFIREs可能对研究细胞内信号具有广泛的适用性。

据介绍，环状腺苷单磷酸酯（cAMP）信号整合了来自不同的G蛋白偶联受体（如神经调节器受体）的信息，以细胞特异性和亚细胞特异性的方式调节关键的生物过程。然而，cAMP动力学的体内细胞分辨率成像仍然具有挑战性。

附：英文原文

Title: Sensitive genetically encoded sensors for population and subcellular imaging of cAMP in vivo

Author: Massengill, Crystian I., Bayless-Edwards, Landon, Ceballos, Cesar C., Cebul, Elizabeth R., Cahill, James, Bharadwaj, Arpita, Wilson, Evan, Qin, Maozhen, Whorton, Matthew R., Baconguis, Isabelle, Ye, Bing, Mao, Tianyi, Zhong, Haining

Issue&Volume: 2022-10-27

Abstract: Cyclic adenosine monophosphate (cAMP) signaling integrates information from diverse G-protein-coupled receptors, such as neuromodulator receptors, to regulate pivotal biological processes in a cellular-specific and subcellular-specific manner. However, in vivo cellular-resolution imaging of cAMP dynamics remains challenging. Here, we screen existing genetically encoded cAMP sensors and further develop the best performer to derive three improved variants, called cAMPFIREs. Compared with their parental sensor, these sensors exhibit up to 10-fold increased sensitivity to cAMP and a cytosolic distribution. cAMPFIREs are compatible with both ratiometric and fluorescence lifetime imaging and can detect cAMP dynamics elicited by norepinephrine at physiologically relevant, nanomolar concentrations. Imaging of cAMPFIREs in awake mice reveals tonic levels of cAMP in cortical neurons that are associated with wakefulness, modulated by opioids, and differentially regulated across subcellular compartments. Furthermore, enforced locomotion elicits neuron-specific, bidirectional cAMP dynamics. cAMPFIREs also function in Drosophila. Overall, cAMPFIREs may have broad applicability for studying intracellular signaling in vivo.

DOI: 10.1038/s41592-022-01646-5

Source: https://www.nature.com/articles/s41592-022-01646-5

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex