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A fast genetically encoded fluorescent sensor for faithful in vivo acetylcholine detection in mice, fish, worms and flies

Philip M. BordeniD, Peng ZhangiD, Amol V. ShivangeiD, Jonathan S. MarviniD, Joseph CichoniD, Chuntao DaniD, Kaspar PodgorskiiD, António Manuel Pereira De FigueiredoiD, Ondřej NovákiD, Masashi TanimotoiD, Eiji ShigetomiiD, Mark A. Lobas, Hyun-Tae KimiD, Paula K. ZhuiD, Yajun ZhangiD, Wan ZhengiD, Chengcheng FaniD, Guangfu WangiD, Bowen Xiang, Li Gan, Guang‐Xian ZhangiD, Kaiming GuoiD, Lin LiiD, Yuan Cai, Andrew G. YeeiD, , Loren L. LoogeriD

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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 animals.

Abstract summary

Claude Opus 5.5 · xhigh effort · 18 Oct 2026 · from the abstract · CC BY-NC-ND 4.0

iAChSnFR, a fluorescent indicator built from a bacterial periplasmic binding protein, reports acetylcholine with large, fast signals in mouse, fish, fly and worm preparations.

Acetylcholine is a ubiquitous neurotransmitter, and this work engineers a genetically encoded fluorescent indicator for it from a bacterial periplasmic binding protein. The authors report large signal changes, fast rise and decay, little sensitivity to most cholinergic drugs, and detection of transients in four model organisms.

Key results

  1. ClaimedThe engineered indicator iAChSnFR is reported to combine a large fluorescence change on acetylcholine binding with rapid rise and decay kinetics.

    Stated qualitatively in the abstract; no dynamic range, affinity or time constants are given in this text. · Abstract

  2. ClaimedThe sensor is described as insensitive to most cholinergic drugs, which would let it be used alongside pharmacological manipulation of cholinergic signalling.

    Asserted without naming the compounds tested or the residual responses in this text. · Abstract

  3. ClaimedLarge acetylcholine transients were detected with the sensor in slice and in vivo preparations in four animal groups: mouse, fish, fly and worm.

    Reported as an observation across a variety of preparations; no amplitudes, recording conditions or sample numbers appear in this text. · Abstract

Methods, data and reproducibility

Design and optimisation of a genetically encoded fluorescent acetylcholine indicator derived from a bacterial periplasmic binding protein, followed by characterisation of its response amplitude, kinetics and drug sensitivity and by imaging in slice and in vivo preparations of several species.

  • Slice and in vivo preparations imaged with the acetylcholine indicator · mouse, fish, fly, worm
Methods & tools
iAChSnFR
Limitations (2)
  • No quantitative characterisation (fluorescence change, affinity, kinetic constants) appears in the available text, so the sensor cannot be compared with other acetylcholine indicators. · noted in this summary
  • Which cholinergic compounds were tested, and which ones the sensor does respond to, is left unspecified. · noted in this summary

Read this if you image neurotransmitter dynamics and want an acetylcholine sensor usable across model organisms and alongside cholinergic drugs.

Citations
97

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