An optimized acetylcholine sensor for monitoring in vivo cholinergic activity.

An optimized acetylcholine sensor for monitoring in vivo cholinergic activity.
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用于监测体内胆碱能活动的优化乙酰胆碱传感器

DOI:
10.1038/s41592-020-0953-2
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发表时间:
2020-11
期刊:
影响因子:
48
通讯作者:
Li Y
Li Y
中科院分区:
生物学1区
文献类型:
--
作者:
Jing M;Li Y;Zeng J;Huang P;Skirzewski M;Kljakic O;Peng W;Qian T;Tan K;Zou J;Trinh S;Wu R;Zhang S;Pan S;Hires SA;Xu M;Li H;Saksida LM;Prado VF;Bussey TJ;Prado MAM;Chen L;Cheng H;Li Y

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直接测量乙酰胆碱(ACh)释放的能力是了解其生理功能的重要一步。在这里,我们优化了GRABACh(GPCR-激活乙酰胆碱)传感器,以实现大幅提高ACh检测的灵敏度,以及减少下游耦合到细胞内途径。改进后的ACh传感器保留了其前身的亚秒级响应动力学、生理相关亲和力和精确的ACh分子特异性。利用这种传感器,我们揭示了隔室乙酰胆碱信号在嗅觉中枢的转基因苍蝇响应外部刺激,包括气味和身体休克。使用光纤光度记录和双光子成像,我们的ACh传感器还能够灵敏地检测小鼠多种行为中多个脑区的单次试验ACh动态。
The ability to directly measure acetylcholine (ACh) release is an essential step towards understanding its physiological function. Here, we optimized the GRABACh (GPCR-Activation‒Based-ACh) sensor to achieve substantially improved sensitivity in ACh detection, as well as reduced downstream coupling to intracellular pathways. The improved version of the ACh sensor retains the sub-second response kinetics, physiological-relevant affinity and precise molecular specificity to ACh of its predecessor. Using this sensor, we revealed compartmental ACh signals in the olfactory center of transgenic flies in response to external stimuli including odor and body shock. Using fiber photometry recording and two-photon imaging, our ACh sensor also enabled sensitive detection of single-trial ACh dynamics in multiple brain regions in mice in a variety of behaviors.
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