Red-shifted GRAB Acetylcholine Sensors for Multiplex Imaging

Overview

Journal bioRxiv

Date 2025 Jan 7

PMID 39763957

Authors

Shu Xie

Xiaolei Miao

Guochuan Li

Yu Zheng

Mengyao Li

En Ji

Jinxu Wang

Shaochuang Li

Ruyi Cai

Lan Geng

Jiesi Feng

Changwei Wei

Yulong Li

Affiliations

Soon will be listed here.

Abstract

The neurotransmitter acetylcholine (ACh) is essential in both the central and peripheral nervous systems. Recent studies highlight the significance of interactions between ACh and various neuromodulators in regulating complex behaviors. The ability to simultaneously image ACh and other neuromodulators can provide valuable information regarding the mechanisms underlying these behaviors. Here, we developed a series of red fluorescent G protein-coupled receptor activation-based (GRAB) ACh sensors, with a wide detection range and expanded spectral profile. The high-affinity sensor, rACh1h, reliably detects ACh release in various brain regions, including the nucleus accumbens, amygdala, hippocampus, and cortex. Moreover, rACh1h can be co-expressed with green fluorescent sensors in order to record ACh release together with other neurochemicals in various behavioral contexts using fiber photometry and two-photon imaging, with high spatiotemporal resolution. These new ACh sensors can therefore provide valuable new insights regarding the functional role of the cholinergic system under both physiological and pathological conditions.

References

Sun F, Zhou J, Dai B, Qian T, Zeng J, Li X . Next-generation GRAB sensors for monitoring dopaminergic activity in vivo. Nat Methods. 2020; 17(11):1156-1166. PMC: 7648260. DOI: 10.1038/s41592-020-00981-9. View

Ballinger E, Ananth M, Talmage D, Role L . Basal Forebrain Cholinergic Circuits and Signaling in Cognition and Cognitive Decline. Neuron. 2016; 91(6):1199-1218. PMC: 5036520. DOI: 10.1016/j.neuron.2016.09.006. View

Nagel G, Szellas T, Huhn W, Kateriya S, Adeishvili N, Berthold P . Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc Natl Acad Sci U S A. 2003; 100(24):13940-5. PMC: 283525. DOI: 10.1073/pnas.1936192100. View

Zhou F, Liang Y, Dani J . Endogenous nicotinic cholinergic activity regulates dopamine release in the striatum. Nat Neurosci. 2001; 4(12):1224-9. DOI: 10.1038/nn769. View

Wan Q, Okashah N, Inoue A, Nehme R, Carpenter B, Tate C . Mini G protein probes for active G protein-coupled receptors (GPCRs) in live cells. J Biol Chem. 2018; 293(19):7466-7473. PMC: 5949987. DOI: 10.1074/jbc.RA118.001975. View

Wan J, Peng W, Li X, Qian T, Song K, Zeng J . A genetically encoded sensor for measuring serotonin dynamics. Nat Neurosci. 2021; 24(5):746-752. PMC: 8544647. DOI: 10.1038/s41593-021-00823-7. View

Deng F, Wan J, Li G, Dong H, Xia X, Wang Y . Improved green and red GRAB sensors for monitoring spatiotemporal serotonin release in vivo. Nat Methods. 2024; 21(4):692-702. PMC: 11377854. DOI: 10.1038/s41592-024-02188-8. View

Soreq H, Seidman S . Acetylcholinesterase--new roles for an old actor. Nat Rev Neurosci. 2001; 2(4):294-302. DOI: 10.1038/35067589. View

Schwarz L, Miyamichi K, Gao X, Beier K, Weissbourd B, DeLoach K . Viral-genetic tracing of the input-output organization of a central noradrenaline circuit. Nature. 2015; 524(7563):88-92. PMC: 4587569. DOI: 10.1038/nature14600. View

10.

Jing M, Zhang P, Wang G, Feng J, Mesik L, Zeng J . A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies. Nat Biotechnol. 2018; 36(8):726-737. PMC: 6093211. DOI: 10.1038/nbt.4184. View

11.

Schultz W, Dayan P, Montague P . A neural substrate of prediction and reward. Science. 1997; 275(5306):1593-9. DOI: 10.1126/science.275.5306.1593. View

12.

Zhuo Y, Luo B, Yi X, Dong H, Miao X, Wan J . Improved green and red GRAB sensors for monitoring dopaminergic activity in vivo. Nat Methods. 2023; 21(4):680-691. PMC: 11009088. DOI: 10.1038/s41592-023-02100-w. View

13.

Payne J, Nadel L . Sleep, dreams, and memory consolidation: the role of the stress hormone cortisol. Learn Mem. 2004; 11(6):671-8. PMC: 534695. DOI: 10.1101/lm.77104. View

14.

Peper K, Bradley R, Dreyer F . The acetylcholine receptor at the neuromuscular junction. Physiol Rev. 1982; 62(4 Pt 1):1271-340. DOI: 10.1152/physrev.1982.62.4.1271. View

15.

Threlfell S, Lalic T, Platt N, Jennings K, Deisseroth K, Cragg S . Striatal dopamine release is triggered by synchronized activity in cholinergic interneurons. Neuron. 2012; 75(1):58-64. DOI: 10.1016/j.neuron.2012.04.038. View

16.

Hasselmo M . The role of acetylcholine in learning and memory. Curr Opin Neurobiol. 2006; 16(6):710-5. PMC: 2659740. DOI: 10.1016/j.conb.2006.09.002. View

17.

Cantu D, Wang B, W Gongwer M, He C, Goel A, Suresh A . EZcalcium: Open-Source Toolbox for Analysis of Calcium Imaging Data. Front Neural Circuits. 2020; 14:25. PMC: 7244005. DOI: 10.3389/fncir.2020.00025. View

18.

Dana H, Mohar B, Sun Y, Narayan S, Gordus A, Hasseman J . Sensitive red protein calcium indicators for imaging neural activity. Elife. 2016; 5. PMC: 4846379. DOI: 10.7554/eLife.12727. View

19.

Krok A, Maltese M, Mistry P, Miao X, Li Y, Tritsch N . Intrinsic dopamine and acetylcholine dynamics in the striatum of mice. Nature. 2023; 621(7979):543-549. PMC: 11577287. DOI: 10.1038/s41586-023-05995-9. View

20.

Jones B . Arousal and sleep circuits. Neuropsychopharmacology. 2019; 45(1):6-20. PMC: 6879642. DOI: 10.1038/s41386-019-0444-2. View