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A Fast Genetically Encoded Fluorescent Sensor for Faithful  in vivo Acetylcholine Detection in Mice, Fish, Worms and Flies

99 Pages Posted: 30 Mar 2020 Sneak Peek Status: Review Complete

See all articles by Philip M. Borden

Philip M. Borden

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Peng Zhang

University of Virginia - Department of Pharmacology

Amol V Shivange

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Jonathan S. Marvin

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Joseph Cichon

New York University (NYU) - Skirball Institute of Biomolecular Medicine

Chuntao Dan

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Kaspar Podgorski

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Antonio Figueiredo

University of Maryland - Department of Anatomy and Neurobiology

Ondrej Novak

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Masashi Tanimoto

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Eiji Shigetomi

University of Yamanashi - Interdisciplinary Graduate School of Medicine

Mark A. Lobas

University of California, Los Angeles (UCLA) - Department of Physiology

Hyuntae Kim

University Clinic Bonn - Department of Neurosurgery

Paula Zhu

Albemarle High School - Math, Engineering & Science Academy

Yajun Zhang

University of Virginia - Department of Pharmacology

W. Sharon Zheng

University of Virginia - Department of Pharmacology

Chengcheng Fan

California Institute of Technology - Division of Chemistry and Chemical Engineering

Guangfu Wang

University of Virginia - Department of Pharmacology

Bowen Xiang

University of Virginia - Department of Pharmacology

Li Gan

Weill Cornell Medicine

Guang-Xian Zhang

Thomas Jefferson University - Department of Neurology

Kaiming Guo

University of Virginia - Department of Pharmacology

Li Lin

Wenzhou Medical University - School of Pharmaceutical Sciences

Yuan Cai

University of Colorado, Aurora - Department of Pharmacology

Andrew G. Yee

University of Colorado, Aurora - Department of Pharmacology

Abhi Aggarwal

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Huan Bao

University of Wisconsin - Madison - Department of Neuroscience

Xiaochu Lou

University of Wisconsin - Madison - Department of Neuroscience

Edwin R. Chapman

University of Wisconsin - Madison - Department of Neuroscience

Christopher P. Ford

University of Colorado, Aurora - Department of Pharmacology

Douglas Rees

California Institute of Technology - Division of Chemistry and Chemical Engineering

Dirk Dietrich

University Clinic Bonn - Department of Neurosurgery

Baljit S. Khakh

University of California, Los Angeles (UCLA) - Department of Physiology

Jeremy S. Dittman

Weill Cornell Medicine

Wen-Biao Gan

New York University (NYU) - Skirball Institute of Biomolecular Medicine

Minoru Koyama

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Vivek Jayaraman

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

Joseph F. Cheer

University of Maryland - Department of Anatomy and Neurobiology

Henry A. Lester

California Institute of Technology - Division of Biology and Biological Engineering

J. Julius Zhu

University of Virginia - Department of Pharmacology

Loren Looger

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

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Abstract

Here we design and optimize a genetically encoded fluorescent indicator, iAChSnFR, for the ubiquitous neurotransmitter acetylcholine, based on a bacterial periplasmic binding protein. iAChSnFR shows large fluorescence changes, rapid rise and decay kinetics, and insensitivity to most cholinergic drugs. iAChSnFR revealed large transients in a variety of slice and  in vivo preparations in mouse, fish, fly and worm. iAChSnFR will be useful for the study of acetylcholine in all organisms.

Keywords: acetylcholine, functional imaging, genetically encoded sensor, neurotransmitter imaging, fluorescence imaging, in vivo imaging, protein engineering, mouse, fish, Drosophila, C. elegans

Suggested Citation

Borden, Philip M. and Zhang, Peng and Shivange, Amol V and Marvin, Jonathan S. and Cichon, Joseph and Dan, Chuntao and Podgorski, Kaspar and Figueiredo, Antonio and Novak, Ondrej and Tanimoto, Masashi and Shigetomi, Eiji and Lobas, Mark A. and Kim, Hyuntae and Zhu, Paula and Zhang, Yajun and Zheng, W. Sharon and Fan, Chengcheng and Wang, Guangfu and Xiang, Bowen and Gan, Li and Zhang, Guang-Xian and Guo, Kaiming and Lin, Li and Cai, Yuan and Yee, Andrew G. and Aggarwal, Abhi and Bao, Huan and Lou, Xiaochu and Chapman, Edwin R. and Ford, Christopher P. and Rees, Douglas and Dietrich, Dirk and Khakh, Baljit S. and Dittman, Jeremy S. and Gan, Wen-Biao and Koyama, Minoru and Jayaraman, Vivek and Cheer, Joseph F. and Lester, Henry A. and Zhu, J. Julius and Looger, Loren, A Fast Genetically Encoded Fluorescent Sensor for Faithful  in vivo Acetylcholine Detection in Mice, Fish, Worms and Flies. Available at SSRN: https://ssrn.com/abstract=3554080 or http://dx.doi.org/10.2139/ssrn.3554080
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Philip M. Borden

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus ( email )

19700 Helix Drive
Ashburn, VA 20147
United States

Peng Zhang (Contact Author)

University of Virginia - Department of Pharmacology

1400 University Ave
Charlottesville, VA 22903
United States

Amol V Shivange

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus ( email )

19700 Helix Drive
Ashburn, VA 20147
United States

Jonathan S. Marvin

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

19700 Helix Drive
Ashburn, VA 20147
United States

Joseph Cichon

New York University (NYU) - Skirball Institute of Biomolecular Medicine ( email )

540 First Avenue
New York, NY 10016
United States

Chuntao Dan

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus ( email )

19700 Helix Drive
Ashburn, VA 20147
United States

Kaspar Podgorski

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus ( email )

19700 Helix Drive
Ashburn, VA 20147
United States

Antonio Figueiredo

University of Maryland - Department of Anatomy and Neurobiology

Baltimore, MD
United States

Ondrej Novak

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus ( email )

19700 Helix Drive
Ashburn, VA 20147
United States

Masashi Tanimoto

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus ( email )

19700 Helix Drive
Ashburn, VA 20147
United States

Eiji Shigetomi

University of Yamanashi - Interdisciplinary Graduate School of Medicine

Yamanashi
Japan

Mark A. Lobas

University of California, Los Angeles (UCLA) - Department of Physiology ( email )

Los Angeles, CA
United States

Hyuntae Kim

University Clinic Bonn - Department of Neurosurgery

Bonn
Germany

Paula Zhu

Albemarle High School - Math, Engineering & Science Academy

Charlottesville, VA
United States

Yajun Zhang

University of Virginia - Department of Pharmacology

1400 University Ave
Charlottesville, VA 22903
United States

W. Sharon Zheng

University of Virginia - Department of Pharmacology ( email )

1400 University Ave
Charlottesville, VA 22903
United States

Chengcheng Fan

California Institute of Technology - Division of Chemistry and Chemical Engineering ( email )

1200 East California Boulevard
Pasadena, CA 91125
United States

Guangfu Wang

University of Virginia - Department of Pharmacology ( email )

1400 University Ave
Charlottesville, VA 22903
United States

Bowen Xiang

University of Virginia - Department of Pharmacology ( email )

1400 University Ave
Charlottesville, VA 22903
United States

Li Gan

Weill Cornell Medicine

1300 York Avenue
New York, NY 10065
United States

Guang-Xian Zhang

Thomas Jefferson University - Department of Neurology

United States

Kaiming Guo

University of Virginia - Department of Pharmacology

1400 University Ave
Charlottesville, VA 22903
United States

Li Lin

Wenzhou Medical University - School of Pharmaceutical Sciences

Wenzhou
China

Yuan Cai

University of Colorado, Aurora - Department of Pharmacology ( email )

Aurora, CO
United States

Andrew G. Yee

University of Colorado, Aurora - Department of Pharmacology

Aurora, CO
United States

Abhi Aggarwal

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus ( email )

19700 Helix Drive
Ashburn, VA 20147
United States

Huan Bao

University of Wisconsin - Madison - Department of Neuroscience

1111 Highland Ave.
Room 5505 WIMR-II
Madison, WI 53705
United States

Xiaochu Lou

University of Wisconsin - Madison - Department of Neuroscience

1111 Highland Ave.
Room 5505 WIMR-II
Madison, WI 53705
United States

Edwin R. Chapman

University of Wisconsin - Madison - Department of Neuroscience

1111 Highland Avenue
Room 5505 WIMR-II
Madison, WI 53705
United States

Christopher P. Ford

University of Colorado, Aurora - Department of Pharmacology

Aurora, CO
United States

Douglas Rees

California Institute of Technology - Division of Chemistry and Chemical Engineering ( email )

1200 East California Boulevard
Pasadena, CA 91125
United States

Dirk Dietrich

University Clinic Bonn - Department of Neurosurgery ( email )

Bonn
Germany

Baljit S. Khakh

University of California, Los Angeles (UCLA) - Department of Physiology ( email )

Los Angeles, CA
United States

Jeremy S. Dittman

Weill Cornell Medicine ( email )

1300 York Avenue
New York, NY 10065
United States

Wen-Biao Gan

New York University (NYU) - Skirball Institute of Biomolecular Medicine ( email )

540 First Avenue
New York, NY 10016
United States

Minoru Koyama

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

19700 Helix Drive
Ashburn, VA 20147
United States

Vivek Jayaraman

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus

19700 Helix Drive
Ashburn, VA 20147
United States

Joseph F. Cheer

University of Maryland - Department of Anatomy and Neurobiology

Baltimore, MD
United States

Henry A. Lester

California Institute of Technology - Division of Biology and Biological Engineering

Pasadena, CA 91125
United States

J. Julius Zhu

University of Virginia - Department of Pharmacology ( email )

1400 University Ave
Charlottesville, VA 22903
United States

Loren Looger

Howard Hughes Medical Institute (HHMI) - Chevy Chase - Janelia Research Campus ( email )

19700 Helix Drive
Ashburn, VA 20147
United States

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