Genetically Engineered Fluorescent Voltage Reporters

Mutoh, Hiroki; Akemann, Walther; Knöpfel, Thomas

doi:10.1021/cn300041b

Cited by 54 publications

(42 citation statements)

References 46 publications

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“…A major limitation of FACILE imaging of calcium is poor temporal resolution compared with electrophysiology. However, in the future, we could avoid the inherently slow calcium response by using voltage-sensitive indicators (Mutoh et al 2012).…”

Section: Discussionmentioning

confidence: 99%

Imaging light responses of retinal ganglion cells in the living mouse eye

Yin

Geng

Osakada

et al. 2013

Journal of Neurophysiology

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This study reports development of a novel method for high-resolution in vivo imaging of the function of individual mouse retinal ganglion cells (RGCs) that overcomes many limitations of available methods for recording RGC physiology. The technique combines insertion of a genetically encoded calcium indicator into RGCs with imaging of calcium responses over many days with FACILE (functional adaptive optics cellular imaging in the living eye). FACILE extends the most common method for RGC physiology, in vitro physiology, by allowing repeated imaging of the function of each cell over many sessions and by avoiding damage to the retina during removal from the eye. This makes it possible to track changes in the response of individual cells during morphological development or degeneration. FACILE also overcomes limitations of existing in vivo imaging methods, providing fine spatial and temporal detail, structure-function comparison, and simultaneous analysis of multiple cells.retinal ganglion cells; in vivo adaptive optics imaging; calcium imaging AVAILABLE METHODS FOR EXAMINING the physiology of retinal ganglion cells (RGCs) have marked limitations, which are resolved by the novel in vivo imaging method described here.

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Section: Discussionmentioning

confidence: 99%

Imaging light responses of retinal ganglion cells in the living mouse eye

Yin

Geng

Osakada

et al. 2013

Journal of Neurophysiology

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“…12 Two recently developed GEVI proteins, ArcLight 16 and MacQ, 17 can detect multiple-spike-averaged fluorescence responses with time resolution in the tens of milliseconds. Another new probe, ASAP1, which is based on circularly permuted GFP, generates responses 1 to 10 ms long in response to single stimulated action potentials in cultured human embryonic kidney cells.…”

Section: Neural Activity-dependent Probes: Past Present and Near mentioning

confidence: 99%

Fast calcium sensor proteins for monitoring neural activity

et al. 2014

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A major goal of the BRAIN Initiative is the development of technologies to monitor neuronal network activity during active information processing. Toward this goal, genetically encoded calcium indicator proteins have become widely used for reporting activity in preparations ranging from invertebrates to awake mammals. However, slow response times, the narrow sensitivity range of Ca2+ and in some cases, poor signal-to-noise ratio still limit their usefulness. Here, we review recent improvements in the field of neural activity-sensitive probe design with a focus on the GCaMP family of calcium indicator proteins. In this context, we present our newly developed Fast-GCaMPs, which have up to 4-fold accelerated off-responses compared with the next-fastest GCaMP, GCaMP6f. Fast-GCaMPs were designed by destabilizing the association of the hydrophobic pocket of calcium-bound calmodulin with the RS20 binding domain, an intramolecular interaction that protects the green fluorescent protein chromophore. Fast-GCaMP6f-RS06 and Fast-GCaMP6f-RS09 have rapid off-responses in stopped-flow fluorimetry, in neocortical brain slices, and in the intact cerebellum in vivo. Fast-GCaMP6f variants should be useful for tracking action potentials closely spaced in time, and for following neural activity in fast-changing compartments, such as axons and dendrites. Finally, we discuss strategies that may allow tracking of a wider range of neuronal firing rates and improve spike detection.

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“…• Scale electromagnetic reading, writing and blocking of action potentials to highlyparallel nerve fibres o Extend the optogenetic toolbox to the periphery with the aim of imaging a population of individual action potentials and introducing or inhibiting distributed naturalistic signalling patterns [39][40][41] o Establish the in vivo reliability, temporal and spatial resolution of micro-and nano-particle reporters and actuators that can be remotely interrogated by optical or electromagnetic imaging approaches (for example, nano-dots, nano-diamonds, magnetic-and piezo-electric particles ("neural dust")) [41][42][43][44] o Develop methods for delivery, expression, and, distribution of nano-particle and genetically-encoded reporters into peripheral nerve fibres 45 …”

Section: Research Imperativesmentioning

confidence: 99%

Bioelectronic medicines: a research roadmap

Birmingham

Gradinaru

Anikeeva

et al. 2014

Nat Rev Drug Discov

303

248

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In format provided by Birmingham et al. (JUNE 2014) NATURE REVIEWS | DRUG DISCOVERY www.nature.com/reviews/drugdisc Box S1 | Bioelectronic medicines: the detailed research roadmap Creation of a visceral nerve atlas Structural mappingOverall objective: Create organ-centric wiring diagrams in models representative of human anatomy. Research imperatives:• Generate tools for high-resolution tracing of the fibre anatomy and taxonomy to and from individual organs o Build a library of tracers to visualize the full length of pre-ganglionic axons, ganglionic cell bodies, post-ganglionic axons, and intrinsic neurons 1 , with particular focus on tracing that starts at the target organ 2-5 o Advance approaches for high-resolution labelling of peripheral neurotransmitters, their receptors and co-receptors, and for imaging of myelination, peri-and epineurium o Develop and adapt micrometer-resolution imaging and 3D reconstruction techniques for visceral organs and peripheral nerves (for example, automated tissue slicing, clearing, in situ hybridization, multi-photon imaging) 6-12 • Explore inter-and intra-species variation in neuroanatomy and establish the optimal animal models for detailed mapping of each organ o Update and extend the macro-level innervation map of the major visceral organs in key animal model species and establish the extent to which this map is conserved in humans o Characterise the variability in different parts of these maps between individuals • Build organ-centric high-resolution maps for each visceral organ in their most representative animal model o Conduct high-resolution nerve tracing, labelling, and imaging in the animal model of choice, taking the organ as the starting point o Standardise and coordinate mapping, data management and 3D visualisation across organs • Establish methods to image and find nerves in the clinical setting o Develop tracers and associated imaging techniques that can be used in human preoperative and intra-operative settings to identify and localize peripheral nerves o Identify anatomical landmarks associated with putative intervention points for surgery Functional mappingOverall objective: Map the neural signalling patterns that control individual organ functions. Research imperatives:• Generate simultaneous recordings of neural signal pattern and organ function o Record both afferent (sensory) and efferent (motor) neural signals and associated end-organ biomarkers at a range of physiological stimuli

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Genetically Engineered Fluorescent Voltage Reporters

Cited by 54 publications

References 46 publications

Imaging light responses of retinal ganglion cells in the living mouse eye

Imaging light responses of retinal ganglion cells in the living mouse eye

Fast calcium sensor proteins for monitoring neural activity

Bioelectronic medicines: a research roadmap

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