RT Journal Article
SR Electronic
T1 A genetically-encoded fluorescent acetylcholine indicator
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 311126
DO 10.1101/311126
A1 Miao Jing
A1 Peng Zhang
A1 Guangfu Wang
A1 Huoqing Jiang
A1 Lukas Mesik
A1 Jiesi Feng
A1 Jianzhi Zeng
A1 Shaohua Wang
A1 Jess Looby
A1 Nick A. Guagliardo
A1 Linda W. Langma
A1 Ju Lu
A1 Yi Zuo
A1 David A. Talmage
A1 Lorna W. Role
A1 Paula Q. Barrett
A1 Li I. Zhang
A1 Minmin Luo
A1 Yan Song
A1 J. Julius Zhu
A1 Yulong Li
YR 2018
UL http://biorxiv.org/content/early/2018/05/01/311126.abstract
AB Acetylcholine (ACh) regulates a diverse array of physiological processes throughout the body, yet cholinergic transmission in the majority of tissues/organs remains poorly understood due primarily to the limitations of available ACh-monitoring techniques. We developed a family of G-protein-coupled receptor activation-based ACh sensors (GACh) with sensitivity, specificity, signal-to-noise ratio, kinetics and photostability suitable for monitoring ACh signals in vitro and in vivo. GACh sensors were validated with transfection, viral and/or transgenic expression in a dozen types of neuronal and non-neuronal cells prepared from several animal species. In all preparations, GACh sensors selectively responded to exogenous and/or endogenous ACh with robust fluorescence signals that were captured by epifluorescent, confocal and/or two-photon microscopy. Moreover, analysis of endogenous ACh release revealed firing pattern-dependent release and restricted volume transmission, resolving two long-standing questions about central cholinergic transmission. Thus, GACh sensors provide a user-friendly, broadly applicable toolbox for monitoring cholinergic transmission underlying diverse biological processes.