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A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies.

ABSTRACT: The neurotransmitter acetylcholine (ACh) regulates a diverse array of physiological processes throughout the body. Despite its importance, cholinergic transmission in the majority of tissues and organs remains poorly understood owing primarily to the limitations of available ACh-monitoring techniques. We developed a family of ACh sensors (GACh) based on G-protein-coupled receptors that has the 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 multiple animal species. In all preparations, GACh sensors selectively responded to exogenous and/or endogenous ACh with robust fluorescence signals that were captured by epifluorescence, 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 tool for monitoring cholinergic transmission underlying diverse biological processes.

SUBMITTER: Jing M

PROVIDER: S-EPMC6093211 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature

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A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies.

Jing Miao M Jing Miao M Zhang Peng P Wang Guangfu G Feng Jiesi J Mesik Lukas L Zeng Jianzhi J Jiang Huoqing H Wang Shaohua S Looby Jess C JC Guagliardo Nick A NA Langma Linda W LW Lu Ju J Zuo Yi Y Talmage David A DA Role Lorna W LW Barrett Paula Q PQ Zhang Li I LI Luo Minmin M Song Yan Y Zhu J Julius JJ Li Yulong Y

Nature biotechnology 20180709 8

The neurotransmitter acetylcholine (ACh) regulates a diverse array of physiological processes throughout the body. Despite its importance, cholinergic transmission in the majority of tissues and organs remains poorly understood owing primarily to the limitations of available ACh-monitoring techniques. We developed a family of ACh sensors (GACh) based on G-protein-coupled receptors that has the sensitivity, specificity, signal-to-noise ratio, kinetics and photostability suitable for monitoring AC ...[more]

PMID: 29985477

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Dataset Information

A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies.

Publications

A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies.

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