A genetically encoded fluorescent sensor for in vivo imaging of GABA

Marvin, Jonathan S; Shimoda, Yoshiteru; Magloire, Vincent; Leite, Marco; Kawashima, Takashi; Jensen, Thomas P.; Kolb, Ilya; Knott, Erika L.; Novák, Ondřej; Podgorski, Kaspar; Leidenheimer, Nancy J.; Rusakov, Dmitri A.; Ahrens, Misha B.; Kullmann, Dimitri M.; Looger, Loren L.

doi:10.1038/s41592-019-0471-2

Cited by 285 publications

(214 citation statements)

References 55 publications

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“…Receptor-based sensors also recognize a host of drug molecules, precluding their simultaneous use with imaging. Meanwhile, PBP-based indicators have few such restrictions, which has allowed use of TBOA, ketamine, NBQX, and other drugs with iGluSnFR 24,25,84,85 , vigabatrin and isoflurane with iGABASnFR 26 , and here, tacrine, donepezil, oxotremorine and eserine with iAChSnFR.…”

Section: Discussionmentioning

confidence: 99%

“…These utilize microbial periplasmic binding proteins (PBPs) as the molecular recognition moiety and circularly permuted GFP or its analogs as the reporter moieties. The strategy has created sensors for glutamate (iGluSnFR 24 , SF-iGluSnFR 25 ), GABA (iGABASnFR) 26 , glucose (iGlucoSnFR 27 , iGlucoSnFR-TS 28 ), maltose 29 , and phosphonate 30 . Because of their basis on microbial proteins, they are bio-orthogonal to endogenous signaling pathways in model organisms, permitting both long-term in vivo expression and use alongside most relevant pharmaceuticals.…”

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confidence: 99%

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A fast genetically encoded fluorescent sensor for faithfulin vivoacetylcholine detection in mice, fish, worms and flies

Zhang

Shivange

et al. 2020

Preprint

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138

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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. IntroductionAcetylcholine (ACh) is a critical neurotransmitter in all animals. Among invertebrates, it is the most prevalent excitatory transmitter in the brain, sensory ganglia, and frequently the neuromuscular junction (NMJ). Among vertebrates, only a minority of neurons release ACh, but these signals play varying key roles. For instance, ACh signals at the NMJ, in the autonomic nervous system, and in subsets of the central nervous system, particularly projections arising from the brainstem and basal forebrain. Other cholinergic neuron populations in the brain include striatal interneurons, the stria vascularis-medial habenula-interpeduncular nucleus pathway, and sparse, incompletely characterized cell types such as intrinsic cholinergic interneurons in cortex 1 and hippocampus 2 . ACh helps to regulate attention 3 and wakefulness 4 , and participates in memory formation and consolidation 5 . ACh is also an important transmitter in glia, and between the nervous and immune systems 6 .Acetylcholine is synthesized pre-synaptically from choline and acetyl-CoA by choline acetyltransferase (ChAT), then packaged into synaptic vesicles by the vesicular acetylcholine transporter (VAChT). A key, partially understood aspect of cholinergic signaling is co-release with other neurotransmitters, including GABA, ATP, and glutamate 7,8 . To understand the role of co-release, one must measure ACh release alongside emerging measurements of other neurotransmitters.Acetylcholine receptors are among the most diverse neurotransmitter receptor families. Humans possess five muscarinic G protein-coupled receptors (GPCRs) for ACh (mAChRs) with diverse expression in the brain and smooth, cardiac, and skeletal muscle. Vertebrate nicotinic ACh receptors (nAChRs) are pentameric ligand-gated cation channels. Humans have a total of 17 nAChR subunit genes, in five classes: 10 a, 4 b, and one each of g, d, and e. nAChRs occur with many subunit combinations 9 , and others may be undiscovered. Invertebrates also have AChgated chloride channels. On neurons, receptors can be localized pre-, post-, and extrasynaptically, often with different isoforms in each place 10

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

A fast genetically encoded fluorescent sensor for faithfulin vivoacetylcholine detection in mice, fish, worms and flies

Zhang

Shivange

et al. 2020

Preprint

Self Cite

138

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“…Whether single neurons in cortical circuits have similarly tight spatial and temporal E/I coordination in order to prevent errant dendritic spiking remains to be determined. The recent advent of tools to optically monitor dendritic excitation and inhibition (iGluSnFR (Marvin et al, 2013) and iGABASnFR (Marvin et al, 2019) paves the way for exciting future investigations of how inhibition and excitation shape direction selectivity and other neural computations carried out by diverse circuits in the brain.…”

Section: Discussionmentioning

confidence: 99%

The functional organization of excitation and inhibition in the dendritic arbors of retinal direction-selective ganglion cells

Jain

Murphy-Baum

deRosenroll

et al. 2019

Preprint

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2 SUMMARYRecent studies indicate that the precise timing and location of excitation and inhibition (E/I) within active dendritic trees can significantly impact neuronal function. How excitatory and inhibitory inputs are functionally organized at the subcellular level in intact circuits remains unclear. To address this issue, we took advantage of the retinal direction-selective ganglion cell circuit, in which directionally tuned inhibitory GABAergic input arising from starburst amacrine cells shape direction-selective dendritic responses. We combined two-photon Ca 2+ imaging with genetic, pharmacological, and single-cell ablation methods to examine local E/I. We demonstrate that when active dendritic conductances are blocked, direction selectivity emerges semiindependently within unusually small dendritic segments (<10 µm). Impressively, the direction encoded by each segment is relatively homogenous throughout the ganglion cell's dendritic tree.Together the results demonstrate a precise subcellular functional organization of excitatory and inhibitory input, which suggests that the parallel processing scheme proposed for direction encoding could be more fine-grained than previously envisioned. 3 3

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“…We demonstrate adaptability of the AVATar architecture by designing an agent that detects the neurotransmitter DA. Incorporation of alternative tethered ligands and cognate binding domains could permit a variety of further vasoprobe-based sensors to be constructed, in some cases leveraging wellcharacterized components of previously described fluorescent imaging probes [26][27][28][29][30][31][32][33][34][35] .…”

Section: Discussionmentioning

confidence: 99%

Target-responsive vasoactive probes for ultrasensitive molecular imaging

Ohlendorf

Wiśniowska

Desai

et al. 2019

Preprint

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ABSTRACTThe ability to monitor molecules volumetrically throughout the body could provide valuable biomarkers for studies of healthy function and disease, but noninvasive detection of molecular targets in living subjects often suffers from poor sensitivity or selectivity. Here we describe a family of potent imaging probes that can be activated by molecules of interest in deep tissue, providing a basis for mapping nanomolar-scale analytes without the radiation or heavy metal content associated with traditional molecular imaging agents. The probes are reversibly-caged vasodilators that induce responses detectable by hemodynamic imaging; they are constructed by combining vasoactive peptides with synthetic chemical appendages and protein blocking domains. We use this architecture to create ultrasensitive biotin-responsive imaging agents, which we apply for wide-field mapping of targets in rat brains using functional magnetic resonance imaging. We also adapt the sensor design for detecting the neurotransmitter dopamine, illustrating versatility of this approach for addressing biologically important molecules.

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A genetically encoded fluorescent sensor for in vivo imaging of GABA

Abstract: 149 words) 18

Cited by 285 publications

References 55 publications

A fast genetically encoded fluorescent sensor for faithfulin vivoacetylcholine detection in mice, fish, worms and flies

A fast genetically encoded fluorescent sensor for faithfulin vivoacetylcholine detection in mice, fish, worms and flies

The functional organization of excitation and inhibition in the dendritic arbors of retinal direction-selective ganglion cells

Target-responsive vasoactive probes for ultrasensitive molecular imaging

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