학술논문
An optimized acetylcholine sensor for monitoring in vivo cholinergic activity
Document Type
Report
Author
Jing, Miao; Li, Yuexuan; Zeng, Jianzhi; Huang, Pengcheng; Skirzewski, Miguel; Kljakic, Ornela; Peng, Wanling; Qian, Tongrui; Tan, Ke; Zou, Jing; Trinh, Simon; Wu, Runlong; Zhang, Shichen; Pan, Sunlei; Hires, Samuel A.; Xu, Min; Li, Haohong; Saksida, Lisa M.; Prado, Vania F.; Bussey, Timothy J.; Prado, Marco, A.M.; Chen, Liangyi; Cheng, Heping; Li, Yulong
Source
Nature Methods. November 2020, Vol. 17 Issue 11, p1139, 8 p.
Subject
Language
English
ISSN
1548-7091
Abstract
Author(s): Miao Jing [sup.1] , Yuexuan Li [sup.2] [sup.3] [sup.4] , Jianzhi Zeng [sup.2] [sup.3] [sup.5] , Pengcheng Huang [sup.6] , Miguel Skirzewski [sup.7] , Ornela Kljakic [sup.8] [sup.9] [sup.10] [...]
The ability to directly measure acetylcholine (ACh) release is an essential step toward understanding its physiological function. Here we optimized the GRAB.sub.ACh (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 subsecond response kinetics, physiologically relevant affinity and precise molecular specificity for 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 performing a variety of behaviors. A genetically encoded acetylcholine sensor with improved sensitivity allows detection of cholinergic neurotransmission in vivo in the Drosophila and mouse brain.
The ability to directly measure acetylcholine (ACh) release is an essential step toward understanding its physiological function. Here we optimized the GRAB.sub.ACh (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 subsecond response kinetics, physiologically relevant affinity and precise molecular specificity for 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 performing a variety of behaviors. A genetically encoded acetylcholine sensor with improved sensitivity allows detection of cholinergic neurotransmission in vivo in the Drosophila and mouse brain.