An optimized acetylcholine sensor for monitoring in vivo cholinergic activity

Jing, Miao; Li, Yuexuan; Zeng, Jianzhi; Huang, Pengcheng; Skirzewski, Miguel; Kljakic, Ornela; Peng, Wanling; Qian, Tong; Tan, Ke; Zou, Jing; Trinh, Simon; Wu, Runlong; Zhang, Shichen; Pan, Sunlei; Hires, Samuel Andrew; Xu, Min; Li, Hao‐Hong; Saksida, Lisa M.; Prado, Vânia F.; Bussey, Timothy J.; Chen, Liangyi; Cheng, Heping

doi:10.1038/s41592-020-0953-2

Cited by 245 publications

(210 citation statements)

References 38 publications

(21 reference statements)

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“…Similar tools have recently been developed for directly measuring acetylcholine release, such as GRAB ACh , which would be especially useful in defining the role of acetylcholine in alcohol-associated behaviors across species. 150 …”

Section: Genetic Tools For All Receptors/ion Channelsmentioning

confidence: 99%

Receptors and Channels Associated with Alcohol Use: Contributions from Drosophila

Scaplen

Petruccelli

2021

J Exp Neurosci

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Alcohol Use Disorder (AUD) is a debilitating disorder that manifests as problematic patterns of alcohol use. At the core of AUD’s behavioral manifestations are the profound structural, physiological, cellular, and molecular effects of alcohol on the brain. While the field has made considerable progress in understanding the neuromolecular targets of alcohol we still lack a comprehensive understanding of alcohol’s actions and effective treatment strategies. Drosophila melanogaster is a powerful model for investigating the neuromolecular targets of alcohol because flies model many of the core behavioral elements of AUD and offer a rich genetic toolkit to precisely reveal the in vivo molecular actions of alcohol. In this review, we focus on receptors and channels that are often targeted by alcohol within the brain. We discuss the general roles of these proteins, their role in alcohol-associated behaviors across species, and propose ways in which Drosophila models can help advance the field.

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Section: Genetic Tools For All Receptors/ion Channelsmentioning

confidence: 99%

Receptors and Channels Associated with Alcohol Use: Contributions from Drosophila

Scaplen

Petruccelli

2021

J Exp Neurosci

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“…Genetically encoded ACh sensors include GPCR-based GRAB ACh2.0 and GRAB ACh3.0 [ 24 , 31 ], and PBP-based iAChSnFRs with both green and yellow fluorescent versions [ 29 ] (Table 1 ). Directly comparing the perforamance of GRAB ACh and iAChSnFR sensors in the same medial entorhinal cortical preparation revealed a few key differences between these two families of sensors [ 29 , 32 ].…”

Section: Introductionmentioning

confidence: 99%

“…Directly comparing the perforamance of GRAB ACh and iAChSnFR sensors in the same medial entorhinal cortical preparation revealed a few key differences between these two families of sensors [ 29 , 32 ]. Although GRAB ACh sensors produced robust fluorescence responses, their inherited slow kinetics significantly attenuated and delayed high-frequency cholinergic signals; they were excellent binary cholinergic signal detectors [ 24 , 31 ]. iAChSnFRs performed much better in faithfully following cholinergic signals in the entire frequency range [ 29 , 32 ].…”

Section: Introductionmentioning

confidence: 99%

Genetically encoded sensors enable micro- and nano-scopic decoding of transmission in healthy and diseased brains

et al. 2020

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Neural communication orchestrates a variety of behaviors, yet despite impressive effort, delineating transmission properties of neuromodulatory communication remains a daunting task due to limitations of available monitoring tools. Recently developed genetically encoded neurotransmitter sensors, when combined with superresolution and deconvolution microscopic techniques, enable the first micro- and nano-scopic visualization of neuromodulatory transmission. Here we introduce this image analysis method by presenting its biophysical foundation, practical solutions, biological validation, and broad applicability. The presentation illustrates how the method resolves fundamental synaptic properties of neuromodulatory transmission, and the new data unveil unexpected fine control and precision of rodent and human neuromodulation. The findings raise the prospect of rapid advances in the understanding of neuromodulatory transmission essential for resolving the physiology or pathogenesis of various behaviors and diseases.

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“…1a ). To improve the dynamic range of our eCB sensor, we then selected 8 residues in cpEGFP for individual randomized mutation based our the experience gained through the development of previous GRAB sensors 30,32–34,36–38 ( Extended Data Fig. 1b ).…”

Section: Resultsmentioning

confidence: 99%

A fluorescent sensor for spatiotemporally resolved endocannabinoid dynamics in vitro and in vivo

Dong

Dudok

et al. 2020

Preprint

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Endocannabinoids (eCBs) are retrograde lipid neuromodulators involved in many physiologically important processes. However, their release and dynamics in the brain remain largely unknown, in part due to lack of probes capable of reporting real-time eCBs with sufficient spatiotemporal resolution. Here, we developed a new G protein-coupled receptor Activation Based eCB sensor GRABeCB2.0 using the human CB1 cannabinoid receptor and a circular-permutated EGFP. GRABeCB2.0 exhibited proper cell membrane trafficking, ~seconds kinetics, high specificity and robust fluorescence response to eCBs at physiological concentrations. Using GRABeCB2.0, we detected evoked eCB dynamics in both cultured neurons and acute brain slices. Interestingly, we also observed spontaneous compartmental eCB transients that spread ~11 μm in cultured neurons, suggesting locally-restricted eCB signaling. By expressing GRABeCB2.0in vivo, we readily observed foot-shock elicited and running triggered eCB transients in mouse amygdala and hippocampus, respectively. Lastly, using GRABeCB2.0 in an epilepsy model, we observed a spreading eCB wave following a calcium wave in mouse hippocampus. In summary, GRABeCB2.0 is a powerful new probe to resolve eCB release and dynamics under both physiological and pathological conditions.

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An optimized acetylcholine sensor for monitoring in vivo cholinergic activity

Cited by 245 publications

References 38 publications

Receptors and Channels Associated with Alcohol Use: Contributions from Drosophila

Receptors and Channels Associated with Alcohol Use: Contributions from Drosophila

Genetically encoded sensors enable micro- and nano-scopic decoding of transmission in healthy and diseased brains

A fluorescent sensor for spatiotemporally resolved endocannabinoid dynamics in vitro and in vivo

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