Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators

Chamberland, Simon; Hh, Yang; Pan, Michael; Evans, Stephen W; Guan, Sihui; Chavarha, Mariya; Yang, Ying‐Wei; Salesse, Charleen; Wu, Haodi; Wu, Joseph C.; Clandinin, Thomas R.; Tóth, Katalin; Lin, Michael Z.; St-Pierre, François

doi:10.7554/elife.25690

Cited by 177 publications

(204 citation statements)

References 90 publications

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“…Currently, GEVIs do not provide the same activity contrast ratios as GECIs, but they are able to capture neuronal signals with ~10 ms time resolution (Chamberland et al 2017, Hochbaum et al 2014). We expect that their further development will lead to a synergetic impact with the technological progress on the microscopy side as discussed in this review.…”

Section: Molecular Indicators For Optical Readout Of Neuronal Actimentioning

confidence: 99%

A Guide to Emerging Technologies for Large-Scale and Whole-Brain Optical Imaging of Neuronal Activity

Weisenburger

Vaziri

2018

Annu. Rev. Neurosci.

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The mammalian brain is a densely interconnected network that consists of millions to billions of neurons. Decoding how information is represented and processed by this neural circuitry requires the ability to capture and manipulate the dynamics of large populations at high speed and resolution over a large area of the brain. While there has been a rapid increase in use of optical approaches in the neuroscience community over the last two decades, most microscopy approaches lack the ability to record the activity of all neurons comprising a functional network across the mammalian brain at relevant temporal and spatial resolution. In this review, we survey the recent development in the optical calcium imaging technologies in this regard and provide an overview of the strengths and limitations of each modality and their potential for scalability. We provide a guidance from a biological user perspective that is driven by the typical biological applications and sample conditions. We also discuss the potential for future advances and synergies that could be obtained through hybrid approaches or other modalities.

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Section: Molecular Indicators For Optical Readout Of Neuronal Actimentioning

confidence: 99%

A Guide to Emerging Technologies for Large-Scale and Whole-Brain Optical Imaging of Neuronal Activity

Weisenburger

Vaziri

2018

Annu. Rev. Neurosci.

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“…1a) with one optimized amino acid (aa), resulting in larger responses to hyperpolarization 19 . In ASAP2s, a mutation of Arg-415 to Gln (R415Q, numbering based on contiguity with G. gallus voltage-sensing phosphatase for consistency with ASAP1) improved steady-state responsiveness to depolarization by 66%, but reduced its speed ( Table 1, Supplementary Table 1) 11 . The combined mutant, ASAP2f R414Q (in which aa 414 corresponds to aa 415 in ASAP2s), exhibited responsivity similar to ASAP2s but slightly slower kinetics ( Supplementary Fig.…”

Section: Mechanism-based Evolution Of Asap3mentioning

confidence: 99%

“…Interestingly, ASAP3 preserved the relatively slow return to baseline of ASAP2s, which is useful for prolonging the response to an AP and thereby facilitating the identification of spikes by template matching 11 . At 33 ºC, ASAP2s fluorescence returned to baseline following a 1-ms step in a biexponential manner, with time constants of 2.11 ± 0.40 ms (amplitude 53 ± 10%) and 7.10 ± 1.62 ms (Supplementary Table 2).…”

Section: Detailed Characterization Of Asap3mentioning

confidence: 99%

Fast two-photon volumetric imaging of an improved voltage indicator reveals electrical activity in deeply located neurons in the awake brain

Chavarha

Villette

Dimov

et al. 2018

Preprint

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Imaging of transmembrane voltage deep in brain tissue with cellular resolution has the potential to reveal information processing by neuronal circuits in living animals with minimal perturbation. Multi-photon voltage imaging in vivo, however, is currently limited by speed and sensitivity of both indicators and imaging methods. Here, we report the engineering of an improved genetically encoded voltage indicator, ASAP3, which exhibits up to 51% fluorescence responses in the physiological voltage range, sub-millisecond activation kinetics, and full responsivity under two-photon illumination. We also introduce an ultrafast local volume excitation (ULOVE) two-photon scanning method to sample ASAP3 signals in awake mice at kilohertz rates with increased stability and sensitivity. ASAP3 and ULOVE allowed continuous single-trial tracking of spikes and subthreshold events for minutes in deep locations, with subcellular resolution, and with repeated sampling over multiple days. By imaging voltage in visual cortex neurons, we found evidence for cell type-dependent subthreshold modulation by locomotion. Thus, ASAP3 and ULOVE enable continuous high-speed highresolution imaging of electrical activity in deeply located genetically defined neurons during awake behavior.shaping 7,8 . However, parallel excitation is problematic for imaging, as scattered fluorescence signals from simultaneously excited cells would become intermixed, hindering detection of small responses. Furthermore, as GEVIs reside in the membrane rather than the cytosol, the number of indicator molecules that can be excited in an optical section through a mammalian cell body is typically smaller for GEVIs than for GECIs 4,6 .Given these limitations, the development of GEVIs with larger two-photon responses to electrical events of interest is highly desirable. Currently, the GEVIs with the largest responses to both subthreshold changes and APs are based on two types of voltage-sensing domains: seven-transmembrane helix opsins and four-transemembrane helix voltage-sensing domains (VSDs). Opsin-based GEVIs have been used in vivo with one-photon excitation to report electrical activity of superficially located neurons 9,10 , but their responsivity is severely attenuated under twophoton excitation 4,11 . In contrast, ASAP-family GEVIs, composed of a circularly permuted green fluorescent protein variant inserted within the VSD of G. gallus voltage-sensing phosphatase (Fig. 1a), are fully responsive under twophoton excitation 11 . In particular, ASAP2s demonstrates the largest response per AP of fluorescent protein-based GEVIs, but its kinetics are actually slower than earlier ASAP variants 11 . If ASAP-family kinetics and/or overall responsivity could be improved, then electrical events could be more easily detected by two-photon imaging.Here we report an improved indicator, ASAP3, resulting from novel methods for generation and screening of GEVI libraries in mammalian cells. ASAP3 features the largest responses of fluorescent GEVIs to either steady-state voltages or ...

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“…Due to the desire to be able to image population neural activity with high temporal precision and sensitivity, much recent interest has focused on the development of geneticallyencoded voltage indicators (GEVIs). Near-infrared fluorescent GEVIs derived from rhodopsins offer high temporal fidelity, and are compatible with optogenetics 1-3 , whereas green fluorescent GEVIs derived from voltage sensing domains of phosphatases or opsins are slower and brighter [4][5][6][7][8][9] . Translating these into the living mouse brain has been challenging, because poor membrane localization, photostability, and sensitivity of previous molecules has resulted in poor signal-tonoise ratio (SNR) in vivo.…”

Section: Introductionmentioning

confidence: 99%

Population imaging of neural activity in awake behaving mice in multiple brain regions

Piatkevich

Bensussen²,

Tseng³

et al. 2019

Preprint

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A longstanding goal in neuroscience has been to image membrane voltage, with high temporal precision and sensitivity, in awake behaving mammals. Here, we report a genetically encoded voltage indicator, SomArchon, which exhibits millisecond response times and compatibility with optogenetic control, and which increases the sensitivity, signal-to-noise ratio, and number of neurons observable, by manyfold over previous reagents. SomArchon only requires conventional one-photon microscopy to achieve these high performance characteristics. These improvements enable population analysis of neural activity, both at the subthreshold and spiking levels, in multiple brain regions -cortex, hippocampus, and striatum -of awake behaving mice. Using SomArchon, we detect both positive and negative responses of striatal neurons during movement, highlighting the power of voltage imaging to reveal bidirectional modulation. We also examine how the intracellular subthreshold theta oscillations of hippocampal neurons govern spike output, finding that nearby cells can exhibit highly correlated subthreshold activities, even as they generate highly divergent spiking patterns.

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Fast two-photon imaging of subcellular voltage dynamics in neuronal tissue with genetically encoded indicators

Cited by 177 publications

References 90 publications

A Guide to Emerging Technologies for Large-Scale and Whole-Brain Optical Imaging of Neuronal Activity

A Guide to Emerging Technologies for Large-Scale and Whole-Brain Optical Imaging of Neuronal Activity

Fast two-photon volumetric imaging of an improved voltage indicator reveals electrical activity in deeply located neurons in the awake brain

Population imaging of neural activity in awake behaving mice in multiple brain regions

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