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A genetically-encoded fluorescent acetylcholine indicator

View ORCID ProfileMiao Jing, Peng Zhang, Guangfu Wang, Huoqing Jiang, Lukas Mesik, Jiesi Feng, Jianzhi Zeng, Shaohua Wang, Jess Looby, Nick A. Guagliardo, Linda W. Langma, Ju Lu, Yi Zuo, David A. Talmage, Lorna W. Role, Paula Q. Barrett, Li I. Zhang, Minmin Luo, Yan Song, J. Julius Zhu, Yulong Li
doi: https://doi.org/10.1101/311126
Miao Jing
State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, ChinaPKU-IDG/McGovern Institute for Brain Research, Beijing 100871, ChinaPeking-Tsinghua Center for Life Sciences, Beijing 100871, China
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  • ORCID record for Miao Jing
Peng Zhang
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, VA 22908
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Guangfu Wang
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, VA 22908
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Huoqing Jiang
State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, ChinaPKU-IDG/McGovern Institute for Brain Research, Beijing 100871, ChinaPeking-Tsinghua Center for Life Sciences, Beijing 100871, China
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Lukas Mesik
Zilkha Neurogenetic Institute, Dept. of Physiology & Neuroscience, Keck School of Medicine, University of Southern California, Los Angeles, California, 90033
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Jiesi Feng
State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, ChinaPKU-IDG/McGovern Institute for Brain Research, Beijing 100871, ChinaPeking-Tsinghua Center for Life Sciences, Beijing 100871, China
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Jianzhi Zeng
State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, ChinaPKU-IDG/McGovern Institute for Brain Research, Beijing 100871, ChinaPeking-Tsinghua Center for Life Sciences, Beijing 100871, China
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Shaohua Wang
Department of Neurobiology and Behavior, Stony Brook University, Stony Brook, NY 11794
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Jess Looby
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, VA 22908Undergraduate Class of 2019, University of Virginia College of Arts and Sciences, Charlottesville, VA 22908
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Nick A. Guagliardo
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, VA 22908
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Linda W. Langma
Department of Surgery, University of Virginia School of Medicine, Charlottesville, VA 22908
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Ju Lu
Department of Molecular, Cell, and Developmental Biology, University of California, Santa Cruz, CA 95064
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Yi Zuo
Department of Molecular, Cell, and Developmental Biology, University of California, Santa Cruz, CA 95064
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David A. Talmage
Department of Neurobiology and Behavior, Stony Brook University, Stony Brook, NY 11794
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Lorna W. Role
Department of Neurobiology and Behavior, Stony Brook University, Stony Brook, NY 11794
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Paula Q. Barrett
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, VA 22908
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Li I. Zhang
Zilkha Neurogenetic Institute, Dept. of Physiology & Neuroscience, Keck School of Medicine, University of Southern California, Los Angeles, California, 90033
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Minmin Luo
School of Life Sciences, Tsinghua University, Beijing 100084 ChinaNational Institute of Biological Sciences, Beijing 102206, China
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Yan Song
Peking-Tsinghua Center for Life Sciences, Beijing 100871, China
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J. Julius Zhu
Department of Pharmacology, University of Virginia School of Medicine, Charlottesville, VA 22908
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  • For correspondence: jjzhu@virginia.edu yulongli@pku.edu.cn
Yulong Li
State Key Laboratory of Membrane Biology, Peking University School of Life Sciences, Beijing 100871, ChinaPKU-IDG/McGovern Institute for Brain Research, Beijing 100871, ChinaPeking-Tsinghua Center for Life Sciences, Beijing 100871, China
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  • For correspondence: jjzhu@virginia.edu yulongli@pku.edu.cn
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ABSTRACT

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.

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Posted May 01, 2018.
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A genetically-encoded fluorescent acetylcholine indicator
Miao Jing, Peng Zhang, Guangfu Wang, Huoqing Jiang, Lukas Mesik, Jiesi Feng, Jianzhi Zeng, Shaohua Wang, Jess Looby, Nick A. Guagliardo, Linda W. Langma, Ju Lu, Yi Zuo, David A. Talmage, Lorna W. Role, Paula Q. Barrett, Li I. Zhang, Minmin Luo, Yan Song, J. Julius Zhu, Yulong Li
bioRxiv 311126; doi: https://doi.org/10.1101/311126
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A genetically-encoded fluorescent acetylcholine indicator
Miao Jing, Peng Zhang, Guangfu Wang, Huoqing Jiang, Lukas Mesik, Jiesi Feng, Jianzhi Zeng, Shaohua Wang, Jess Looby, Nick A. Guagliardo, Linda W. Langma, Ju Lu, Yi Zuo, David A. Talmage, Lorna W. Role, Paula Q. Barrett, Li I. Zhang, Minmin Luo, Yan Song, J. Julius Zhu, Yulong Li
bioRxiv 311126; doi: https://doi.org/10.1101/311126

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