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An optimized acetylcholine sensor for monitoring in vivo cholinergic activity
Open accessPublished version of a preprint
Abstract
The ability to directly measure acetylcholine (ACh) release is an essential step toward understanding its physiological function. Here we optimized the GRAB ACh (GPCR-activation-based ACh) sensor to achieve substantially improved sensitivity in ACh detection, as well as reduced downstream coupling to intracellular pathways. The improved version of the ACh sensor retains the subsecond response kinetics, physiologically relevant affinity and precise molecular specificity for ACh of its predecessor. Using this sensor, we revealed compartmental ACh signals in the olfactory center of transgenic flies in response to external stimuli including odor and body shock. Using fiber photometry recording and two-photon imaging, our ACh sensor also enabled sensitive detection of single-trial ACh dynamics in multiple brain regions in mice performing a variety of behaviors.
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Flow fiber photometry enables recordings from deep brain tissues and combined monitoring of blood and CSF dynamics using virally-delivered biosensors.
“flow fiber photometry enables recordings from deep brain tissues and combined monitoring of blood and CSF dynamics using virally-delivered biosensors45,101–104.”
From Norepinephrine-mediated slow vasomotion drives glymphatic clearance during sleep (2025) · cites this paper as [101]
The Online Methods section, “Fiber photometry, EEG and EMG recordings in mice” (page not provided), describes fiber-photometry recordings in the basolateral amygdala and hippocampus using an AAV-delivered ACh sensor. It does not report combined monitoring of blood and CSF dynamics; its mention of hemodynamic change concerns correction of a recording artifact. (LLM, conf=0.87)
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Metadata
- Authors
- Jing, M., Li, Y., Zeng, J., Huang, P., Skirzewski, M., Kljakic, O., Peng, W., Qian, T., Tan, K., Zou, J., Trinh, S., Wu, R., Zhang, S., Pan, S., Hires, S. A., Xu, M., Li, H., Saksida, L. M., Prado, V. F., Bussey, T. J., Prado, M. A. M., Chen, L., Cheng, H., & Li, Y.
- Affiliations
- Chinese Institute for Brain Research (CN)
- Peking University (CN)
- Wuhan National Laboratory for Optoelectronics (CN)
- Western University (CA)
- Chinese Academy of Sciences (CN)
- University of Southern California (US)
- Chinese Academy of Medical Sciences & Peking Union Medical College (CN)
- Corresponding author
- Jing, Miao
- Li, Yulong
- ORCID
- Venue
- Nature Methods
- Published
- 2020-09-28
- Volume
- 17
- Issue
- 11
- Pages
- 1139-1146
- ISSN
- 1548-7091 (ISSN-L)
- 1548-7105
- PMC
- PMC7606762
- OpenAlex
- W3090462036
- Cited by
- 408
- References
- 3
- Citations by year
- 2026: 51 · 2025: 94 · 2024: 79 · 2023: 77 · 2022: 52 · 2021: 42 · 2020: 13
- FWCI
- 20.60
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Locations
- Nature Methods (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Scholarship@Western (Western University) (submittedVersion)
- Funders
- National Natural Science Foundation of China (31671118)
- Peking University
- Canada First Research Excellence Fund
- State Key Laboratory of Membrane Biology
- National Institutes of Health
- Canadian Institutes of Health Research (159781, 159781_1)
- National Institute of Mental Health (R01MH101377)
- National Institute of Neurological Disorders and Stroke (U01NS113358, U01NS103558)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurobiology and Insect Physiology Research
- Keywords
- genetically encoded biosensors, cholinergic neurotransmission, GPCR activation, olfactory center, Drosophila, mouse brain, fiber photometry, two-photon imaging, olfactory response
- MeSH terms
- Acetylcholine, Animals, Behavior, Animal, Brain, Drosophila, Humans, Neurons, Somatosensory Cortex, Biosensing Techniques, Cholinergic Agents, Mushroom Bodies, Animals, Genetically Modified, Receptor, Muscarinic M3, Green Fluorescent Proteins, Mice, HEK293 Cells, Olfactory Cortex
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