Quantitative <i>in vivo</i> imaging of neuronal glucose concentrations with a genetically encoded fluorescence lifetime sensor

Díaz‐García, Carlos Manlio; Lahmann, Carolina; Martínez-François, Juan Ramón; Li, Binsen; Koveal, Dorothy; Nathwani, Nidhi; Rahman, Mahia; Keller, Jacob P.; Marvin, Jonathan S; Looger, Loren L.; Yellen, Gary

doi:10.1002/jnr.24433

Cited by 77 publications

(53 citation statements)

References 60 publications

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“…The probe showed intensiometric response with about 3-fold fluorescence increase after glucose binding and had Kd of 7.7 mM, which is physiologically relevant. iGlucoSnFR-TS [186], also called Sweetie-TS, was created by replacing cpGFP with a pH-stable FP cpT-Sapphire. This probe is considered as a fluorescence lifetime sensor that allows the conversion of measurements into actual concentrations in vivo.…”

Section: Classification Of Cpfp-based Gefis By Analyte Measured Anmentioning

confidence: 99%

Circularly Permuted Fluorescent Protein-Based Indicators: History, Principles, and Classification

Kostyuk

Demidovich

Kotova

et al. 2019

IJMS

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Genetically encoded biosensors based on fluorescent proteins (FPs) are a reliable tool for studying the various biological processes in living systems. The circular permutation of single FPs led to the development of an extensive class of biosensors that allow the monitoring of many intracellular events. In circularly permuted FPs (cpFPs), the original N- and C-termini are fused using a peptide linker, while new termini are formed near the chromophore. Such a structure imparts greater mobility to the FP than that of the native variant, allowing greater lability of the spectral characteristics. One of the common principles of creating genetically encoded biosensors is based on the integration of a cpFP into a flexible region of a sensory domain or between two interacting domains, which are selected according to certain characteristics. Conformational rearrangements of the sensory domain associated with ligand interaction or changes in the cellular parameter are transferred to the cpFP, changing the chromophore environment. In this review, we highlight the basic principles of such sensors, the history of their creation, and a complete classification of the available biosensors.

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Section: Classification Of Cpfp-based Gefis By Analyte Measured Anmentioning

confidence: 99%

Circularly Permuted Fluorescent Protein-Based Indicators: History, Principles, and Classification

Kostyuk

Demidovich

Kotova

et al. 2019

IJMS

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“…It has been suggested that astrocytic glycogen indeed plays an important role in supporting neuronal energy needs; and a key phenomenon in this proposed neuro-metabolic coupling between the two cell types is the capacity of astrocytes to increase their ATP production in response to neuronal activity 6,7,8 . This hypothesis, known as astrocyte-neuron lactate shuttle (ANLS), is still under debate, because other work has provided evidence that neurons may also increase their own rate of glycolysis in response to stimulation 9,10 , reflecting the necessity of further methods and approaches to study neuro-glia interaction.…”

Section: +mentioning

confidence: 99%

Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03<sup>YEMK</sup>

Lerchundi

Kafitz

Färfers

et al. 2019

JoVE

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Neuronal activity in the central nervous system (CNS) evokes a high demand on cellular energy provided by the breakdown of adenosine triphosphate (ATP). A large share of ATP is needed to re-install ion gradients across plasma membranes degraded by electrical signaling of neurons. There is evidence that astrocytes-while not generating fast electrical signals themselves-undergo increased production of ATP in response to neuronal activity and support active neurons by providing energy metabolites to them. The recent development of genetically encoded sensors for different metabolites now enables the study of such metabolic interactions between neurons and astrocytes. Here, we describe a protocol for cell-type specific expression of the ATP-sensitive Fluorescence Resonance Energy Transfer-(FRET-) sensor ATeam1.03 YEMK in organotypic tissue slice cultures of the mouse hippocampus and cortex using adeno-associated viral vectors (AAV). Furthermore, we demonstrate how this sensor can be employed for dynamic measurement of changes in cellular ATP levels in neurons and astrocytes upon increases in extracellular potassium and following induction of chemical ischemia (i.e., an inhibition of cellular energy metabolism).

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“…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%

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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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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Quantitative in vivo imaging of neuronal glucose concentrations with a genetically encoded fluorescence lifetime sensor

Cited by 77 publications

References 60 publications

Circularly Permuted Fluorescent Protein-Based Indicators: History, Principles, and Classification

Circularly Permuted Fluorescent Protein-Based Indicators: History, Principles, and Classification

Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03<sup>YEMK</sup>

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

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