Combined Optogenetic and Chemogenetic Control of Neurons

Berglund, Ken; Tung, Jack K.; Higashikubo, Bryan; Gross, Robert E.; Moore, Christopher I.; Hochgeschwender, Ute

doi:10.1007/978-1-4939-3512-3_14

Cited by 29 publications

(38 citation statements)

References 16 publications

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“…Although these approaches have helped improve the potential volume of tissue illumination, the scalability to human brain and hardware dependency is still unclear or impractical for clinical application. One approach that may address both of these challenges is the use of luminopsins[112–115], whereby photoactivation of optogenetic channels is conferred by bioluminescent proteins ( e.g. luciferase) delivered in the same gene payload, thus tracking exactly with the expressed channels.…”

Section: Future Considerationsmentioning

confidence: 99%

Optogenetic Approaches for Controlling Seizure Activity

2016

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Optogenetics, a technique that utilizes light-sensitive ion channels or pumps to activate or inhibit neurons, has allowed scientists unprecedented precision and control for manipulating neuronal activity. With a clinical need to develop more precise and effective therapies for patients with drug-resistant epilepsy, these tools have recently been explored as a novel treatment for halting seizure activity in various animal models. In this review, we provide a detailed and current summary of these optogenetic approaches and provide a perspective on their future clinical application as a potential neuromodulatory therapy.

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Section: Future Considerationsmentioning

confidence: 99%

Optogenetic Approaches for Controlling Seizure Activity

2016

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“…Stimulation only occurs when the CTZ is injected, producing bioluminescent light through catalysis by the luciferase, resulting in the activation of the opsin. This approach takes advantage of both opto-and chemogenetic concepts by utilizing ion channels for current conduction while activating the channels through the application of a chemical compound, thus allowing non-invasive stimulation and recruitment of all targeted actuators as opposed to only those that can be reached by light from a physical source (8)(9)(10)(11)(12)(13)(14)(15). Here we use LMO3 which consists of slow burn Gaussia luciferase fused to Volvox channelrhodopsin 1.…”

Section: Introductionmentioning

confidence: 99%

Restoring Function After Severe Spinal Cord Injury Through Bioluminescence-Driven Optogenetics

Petersen

Sharkey

Pal

et al. 2019

Preprint

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The ability to manipulate specific neuronal populations of the spinal cord following spinal cord injury (SCI) could prove highly beneficial for rehabilitation in patients through maintaining and strengthening still existing neuronal connections and/or facilitating the formation of new connections. A non-invasive and highly specific approach to neuronal stimulation is bioluminescent-optogenetics (BL-OG), where genetically expressed light emitting luciferases are tethered to light sensitive channelrhodopsins (luminopsins, LMO); neurons are activated by the addition of the luciferase substrate coelenterazine (CTZ). This approach utilizes ion channels for current conduction while activating the channels through application of a small chemical compound, thus allowing non-invasive stimulation and recruitment of all targeted neurons. Rats were transduced in the lumbar spinal cord with AAV2/9 to express the excitatory LMO3 under control of a pan-neuronal or motor neuronspecific promoter. A day after contusion injury of the thoracic spine, rats received either CTZ or vehicle every other day for 2 weeks. Activation of either interneuron or motor neuron populations below the level of injury significantly improved locomotor recovery lasting beyond the time of stimulation. Utilizing histological and gene expression methods we identified neuronal plasticity as a likely mechanism underlying the functional recovery. These findings provide a foundation for a rational approach to spinal cord injury rehabilitation, thereby advancing approaches for functional recovery after SCI.

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“…Novel genetic-and optical-based methods that target specific cell types provide a powerful strategy for probing neural mechanisms that underlie perception and action (Boyden et al, 2005;Zhang et al, 2006;Cardin et al, 2009;Nichols and Roth, 2009;Knopfel et al, 2010;Fenno et al, 2011;Tung et al, 2015;Zhu et al, 2015;Berglund et al, 2016a;Roth, 2016;Kim et al, 2017). Of such approaches, optogenetic and chemogenetic strategies are the most widely employed.…”

Section: Introductionmentioning

confidence: 99%

“…In this approach, binding of an oxidative enzyme (luciferase) to a small light-emitting molecule (luciferin) drives bioluminescence that, in turn, regulates a neighboring opsin (Berglund et al, 2013;Birkner et al, 2014;Tung et al, 2015;Berglund et al, 2016b;Berglund et al, 2016a;Park et al, 2017;Prakash et al, 2018;Tung et al, 2018;Zenchak et al, 2018;Berglund et al, 2019). To ensure proximity of the bioluminescent reaction to the recipient opsin, the luminopsin (LMO) construct was invented, in which a luciferase is linked to the optogenetic element by a short 15 amino acids linker (Berglund et al, 2016a). Here, we use LMO3, a molecule that tethers the slow-burn Gaussia luciferase (sbGLuc) to Volvox Channelrhodopsin-1 (VChR1; Figure 1A), and uses the substrate coelenterazine (CTZ) to generate bioluminescence.…”

Section: Introductionmentioning

confidence: 99%

“…Here, we use LMO3, a molecule that tethers the slow-burn Gaussia luciferase (sbGLuc) to Volvox Channelrhodopsin-1 (VChR1; Figure 1A), and uses the substrate coelenterazine (CTZ) to generate bioluminescence. This single molecule permits both chemogenetic regulation by peripheral injection of a luciferin (Berglund et al, 2013;Birkner et al, 2014;Tung et al, 2015;Berglund et al, 2016a), and local optogenetic regulation by external light application or direct intracortical injection of the luciferin (Tung et al, 2015). The BL-OG strategy has several additional distinctive benefits, including the robust biocompatibility of its components, and photon production that confirms the activator reached its target (e.g., by imaging while administering the drug).…”

Section: Introductionmentioning

confidence: 99%

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The BioLuminescent-OptoGenetic in vivo Response to Coelenterazine is Proportional, Sensitive and Specific in Neocortex

Gomez‐Ramirez

Friedman

et al. 2019

Preprint

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BioLuminescent (BL) light production can modulate neural activity and behavior through coexpressed OptoGenetic (OG) elements, an approach termed 'BL-OG'. Yet, the relationship between BL-OG effects and bioluminescent photon emission has not been characterized in vivo.Further, the degree to which BL-OG effects strictly depend on optogenetic mechanisms driven by bioluminescent photons is unknown. Crucial to every neuromodulation method is whether the activator shows a dynamic concentration range driving robust, selective, and non-toxic effects.We systematically tested the effects of four key components of the BL-OG mechanism (luciferin, oxidized luciferin, luciferin vehicle, and bioluminescence), and compared these against effects induced by the Luminopsin-3 (LMO3) BL-OG molecule, a fusion of slow burn Gaussia luciferase (sbGLuc) and Volvox ChannelRhodopsin-1 (VChR1). We performed combined bioluminescence imaging and electrophysiological recordings while injecting specific doses of Coelenterazine (substrate for sbGluc), Coelenteramide (CTM, the oxidized product of CTZ), or CTZ vehicle. CTZ robustly drove activity in mice expressing LMO3, with photon production proportional to firing rate. In contrast, low and moderate doses of CTZ, CTM, or vehicle did not modulate activity in mice that did not express LMO3. We also failed to find bioluminescence effects on neural activity in mice expressing an optogenetically non-sensitive LMO3 variant. We observed weak responses to the highest dose of CTZ in control mice, but these effects were significantly smaller than those observed in the LMO3 group. These results show that in neocortex in vivo, there is a large CTZ range wherein BL-OG effects are specific to its active chemogenetic mechanism.

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Combined Optogenetic and Chemogenetic Control of Neurons

Cited by 29 publications

References 16 publications

Optogenetic Approaches for Controlling Seizure Activity

Optogenetic Approaches for Controlling Seizure Activity

Restoring Function After Severe Spinal Cord Injury Through Bioluminescence-Driven Optogenetics

The BioLuminescent-OptoGenetic in vivo Response to Coelenterazine is Proportional, Sensitive and Specific in Neocortex

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