Format

Send to

Choose Destination
Nat Biotechnol. 2018 Sep;36(8):726-737. doi: 10.1038/nbt.4184. Epub 2018 Jul 9.

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

Jing M1,2,3, Zhang P4, Wang G4,5, Feng J1,2,3, Mesik L6, Zeng J1,2,3, Jiang H1,2,3, Wang S7, Looby JC4,8, Guagliardo NA4, Langma LW9, Lu J10, Zuo Y10, Talmage DA7, Role LW7, Barrett PQ4, Zhang LI6, Luo M11,12, Song Y3, Zhu JJ4,13,14,15, Li Y1,2,3.

Author information

1
State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing, China.
2
PKU-IDG/McGovern Institute for Brain Research, Beijing, China.
3
Peking-Tsinghua Center for Life Sciences, Beijing, China.
4
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, Virginia, USA.
5
Center for Life Sciences, School of Life Science and Technology, Harbin Institute of Technology, Harbin, China.
6
Zilkha Neurogenetic Institute, Department of Physiology & Neuroscience, Keck School of Medicine, University of Southern California, Los Angeles, California, USA.
7
Department of Neurobiology and Behavior, Stony Brook University, Stony Brook, New York, USA.
8
Undergraduate Class of 2019, University of Virginia College of Arts and Sciences, Charlottesville, Virginia, USA.
9
Department of Surgery, University of Virginia School of Medicine, Charlottesville, Virginia, USA.
10
Department of Molecular, Cell, and Developmental Biology, University of California, Santa Cruz, California, USA.
11
School of Life Sciences, Tsinghua University, Beijing, China.
12
National Institute of Biological Sciences, Beijing, China.
13
School of Medicine, Ningbo University, Ningbo, China.
14
Donders Institute for Brain, Cognition and Behavior, Radboud University Nijmegen, Nijmegen, the Netherlands.
15
Department of Physiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

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.

PMID:
29985477
PMCID:
PMC6093211
DOI:
10.1038/nbt.4184
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center