An orange calcium-modulated bioluminescent indicator for non-invasive activity imaging

Oh, Youjin; Park, Yun‐Hee; Cho, Julia; Wu, Haodi; Paulk, Nicole K.; Liu, Lan Xiang; Kim, Namdoo; Kay, Mark A.; Wu, Joseph C.; Lin, Michael Z.

doi:10.1038/s41589-019-0256-z

“…One solution to improve temporal resolution is to utilize an indicator of real-time neuronal activity, such as the intracellular calcium concentration. Indeed, a calcium-dependent luciferase system, Orange CaMBI, has been reported (Oh et al, 2019). However, this system is based on Monitoring In Vivo Neural Graft Activity After SCI 31 NanoLuc-furimazine and might not be appropriate for neural grafts due to its low substrate permeability through the blood-brain barrier (Edinger et al, 1999;Su et al, 2020).…”

Section: Discussionmentioning

confidence: 99%

Novel System to Monitor In Vivo Neural Graft Activity After Spinal Cord Injury

Ago

¹

,

Nagoshi

²

,

Imaizumi

³

et al. 2021

Preprint

0

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Expectations for neural stem/progenitor cell (NS/PC) transplantation as a treatment for spinal cord injury (SCI) are increasing. However, whether and how grafted cells are incorporated into the host neural circuit and contribute to motor function recovery remain unknown. The aim of this project was to establish a novel non-invasive in vivo imaging system to visualize the activity of neural grafts by which we can simultaneously demonstrate the circuit-level integration between the graft and host, and the contribution of graft neuronal activity to host behaviour. We introduced Akaluc, a newly engineered luciferase, under control of a potent neuronal activity-dependent synthetic promoter, E-SARE, into NS/PCs and engrafted the cells into SCI model mice. Through the use of this system, we reveal that the activity of grafted cells was integrated with host behaviour and driven by host neural circuit inputs. This non-invasive system is expected to help elucidate the therapeutic mechanism of cell transplantation treatment for SCI and determine better therapy techniques that maximize the function of cells in the host circuit.

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“…One solution to improve temporal resolution is to utilize an indicator of real-time neuronal activity, such as the intracellular calcium concentration. Indeed, a calcium-dependent luciferase system, Orange CaMBI, has been reported 32 . However, this system is based on NanoLuc-furimazine and might not be appropriate for neural grafts due to its low substrate permeability through the blood-brain barrier 33,34 .…”

Section: Discussionmentioning

confidence: 99%

Novel System to Monitor In Vivo Neural Graft Activity After Spinal Cord Injury

Ago

¹

,

Nagoshi

²

,

Imaizumi

³

et al. 2021

Preprint

0

View full text Add to dashboard Cite

Expectations for neural stem/progenitor cell (NS/PC) transplantation as a treatment for spinal cord injury (SCI) are increasing. However, whether and how grafted cells are incorporated into the host neural circuit and contribute to motor function recovery remain unknown. The aim of this project was to establish a novel non-invasive in vivo imaging system to visualize the activity of neural grafts by which we can simultaneously demonstrate the circuit-level integration between the graft and host, and the contribution of graft neuronal activity to host behaviour. We introduced Akaluc, a newly engineered luciferase, under control of a potent neuronal activity-dependent synthetic promoter, E-SARE, into NS/PCs and engrafted the cells into SCI model mice. Through the use of this system, we reveal that the activity of grafted cells was integrated with host behaviour and driven by host neural circuit inputs. This non-invasive system is expected to help elucidate the therapeutic mechanism of cell transplantation treatment for SCI and determine better therapy techniques that maximize the function of cells in the host circuit.

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“…This luciferingated version of FLiCRE not only exhibits a steeper calcium dependency compared to the lightgated version of the tool, but it would also only require a non-invasive injection of NanoLuc’s substrate furimazine rather than exogenously delivered blue light through an optical fiber. Promisingly, recent studies have demonstrated the in vivo use of NanoLuc in mice (Oh et al, 2019; Su et al, 2020) and in the brain (Germain-Genevois et al, 2016) to perform bioluminescence imaging. Further engineering and optimization of this luciferin-gated FLiCRE design will be needed to enable non-invasive, molecular recording of integrated calcium activity across the brain.…”

Section: Discussionmentioning

confidence: 99%

A molecular calcium integrator reveals a striatal cell-type driving aversion

Kim

¹

,

Sánchez

²

,

Hoerbelt

³

et al. 2020

Preprint

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The ability to record transient cellular events in the DNA or RNA of cells would enable precise, large-scale analysis, selection, and reprogramming of heterogeneous cell populations. Here we report a molecular technology for stable genetic tagging of cells that exhibit activity-related increases in intracellular calcium concentration (FLiCRE). We used FLiCRE to transcriptionally label activated neural ensembles in the nucleus accumbens of the mouse brain during brief stimulation of aversive inputs. Using single-cell RNA sequencing, we detected FLiCRE transcripts among the endogenous transcriptome, providing simultaneous readout of both cell-type and calcium activation history. We identified a cell-type in the nucleus accumbens activated downstream of long-range excitatory projections. Taking advantage of FLiCRE′s modular design, we expressed an optogenetic channel selectively in this cell-type, and showed that direct recruitment of this otherwise genetically-inaccessible population elicits behavioral aversion. The specificity and minute-resolution of FLiCRE enables molecularly-informed characterization, manipulation, and reprogramming of activated cellular ensembles.

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An orange calcium-modulated bioluminescent indicator for non-invasive activity imaging

Cited by 43 publications

References 44 publications

Novel System to Monitor In Vivo Neural Graft Activity After Spinal Cord Injury

Novel System to Monitor In Vivo Neural Graft Activity After Spinal Cord Injury

Novel System to Monitor In Vivo Neural Graft Activity After Spinal Cord Injury

A molecular calcium integrator reveals a striatal cell-type driving aversion

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