A Designer AAV Variant Permits Efficient Retrograde Access to Projection Neurons

Tervo, D. Gowanlock R.; Hwang, Bum Yeol; Viswanathan, Sarada; Gaj, Thomas; Lavzin, Maria; Ritola, Kimberly; Lindo, Sarah; Michael, Susan; Kuleshova, E. P.; Ojala, David S.; Huang, Cheng Chiu; Gerfen, Charles R.; Schiller, Jackie; Dudman, Joshua T.; Hantman, Adam W.; Looger, Loren L.; Schaffer, David V.; Karpova, Alla Y.

doi:10.1016/j.neuron.2016.09.021

Cited by 1,137 publications

(1,095 citation statements)

References 67 publications

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“…In the present study, the expression of hM4Di was limited to NAc GABAergic neurons, but not GABAergic neurons projecting to the NAc, because the retrograde transport efficacy of the AAV-DJ serotype used in the present study is low. 24) Thus, CNO injections may selectively inhibit NAc GABAergic neurons. On the other hand, in vGAT-ChR2-EYFP mice, all GABAergic neurons in the brain express ChR2.…”

Section: Discussionmentioning

confidence: 99%

Activation of GABAergic Neurons in the Nucleus Accumbens Mediates the Expression of Cocaine-Associated Memory

Zhang

Deyama

Domoto

et al. 2018

Biological & Pharmaceutical Bulletin

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Cocaine-associated environmental cues elicit craving and relapse to cocaine use by recalling the rewarding memory of cocaine. However, the neuronal mechanisms underlying the expression of cocaine-associated memory are not fully understood. Here, we investigated the possible contribution of γ-aminobutyrate (GABA) ergic neurons in the nucleus accumbens (NAc), a key brain region associated with the rewarding and reinforcing effects of cocaine, to the expression of cocaine-associated memory using the conditioned place preference ( Key words nucleus accumbens; cocaine; conditioned place preference; designer receptors exclusively activated by designer drugs; γ-aminobutyrate (GABA); mouse Cocaine addiction is a chronic relapsing disease characterized by compulsive drug seeking and taking. The rewarding and reinforcing properties of cocaine are readily associated with the environmental contexts where cocaine is experienced. The cocaine-associated environmental cues elicit craving and relapse to cocaine use by recalling the rewarding memory of cocaine. However, the mechanisms by which environmental cues induce the expression of cocaine-associated memory are not fully understood. To investigate these mechanisms, the conditioned place preference (CPP) test, a widely used Pavlovian conditioning paradigm is considered to be useful. 1) In the conditioning session of the CPP, animals learn to associate the rewarding effects of cocaine and a neutral environmental context that has been paired with cocaine administration by investigators. Thereafter, in the posttest session, animals exhibit a preference for the cocaine-paired environment, likely due to the expression of cocaine-associated memory. Although previous studies revealed that the mesocorticolimbic reward system consisting of the ventral tegmental area (VTA), medial prefrontal cortex (mPFC), and nucleus accumbens (NAc) is involved in the expression of cocaine-associated memory, 2-4) the precise neuronal and circuit mechanisms underlying this behavior remain unclear.The NAc plays an important role in the expression of cocaine-seeking behavior and cocaine-associated memory. [5][6][7] Recent studies have shown that activation of glutamatergic projections from the prelimbic subregion (PL) of the mPFC to the NAc core is required for cue-induced reinstatement of cocaine seeking. 8,9) In addition, intra-NAc infusion of an AMPA receptor antagonist was found to block the expression of cocaine CPP. 2) These findings suggest that excitation of the NAc is involved in the expression of cocaine-associated memory. Considering that the vast majority of NAc neurons are γ-aminobutyrate (GABA) ergic medium spiny neurons (MSNs), 10,11) these GABAergic neurons are likely to contribute to the expression of cocaine CPP. Thus, in the present study, we addressed this issue using designer receptors exclusively activated by designer drugs (DREADD) technology. 12) We selectively expressed G i/o -coupled inhibitory DREADD hM4Di in NAc GABAergic neurons of vesicular GABA transporterCre (vGAT-Cre) mic...

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Section: Discussionmentioning

confidence: 99%

Activation of GABAergic Neurons in the Nucleus Accumbens Mediates the Expression of Cocaine-Associated Memory

Zhang

Deyama

Domoto

et al. 2018

Biological & Pharmaceutical Bulletin

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“…Fluorescent reporters are commonly used for cell type-specific mapping and phenotyping 2,38,39 (Figs. 2-4).…”

Section: Applications Of the Methodsmentioning

confidence: 99%

Widespread and targeted gene expression by systemic AAV vectors: Production, purification, and administration

Challis

Kumar

Chan

et al. 2018

Preprint

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We recently developed novel AAV capsids for efficient and noninvasive gene transfer across the central and peripheral nervous systems. In this protocol, we describe how to produce and systemically administer AAV-PHP viruses to label and/or genetically manipulate cells in the mouse nervous system and organs including the heart. The procedure comprises three separate stages: AAV production, intravenous delivery, and evaluation of transgene expression.The protocol spans eight days, excluding the time required to assess gene expression, and can be readily adopted by laboratories with standard molecular and cell culture capabilities. We provide guidelines for experimental design and choosing the capsid, cargo, and viral dose appropriate for the experimental aims. The procedures outlined here are adaptable to diverse biomedical applications, from anatomical and functional mapping to gene expression, silencing, and editing.

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“…Whether the variable sensitivity observed is due strictly to variations in expression and trafficking of opsins to the axonal membrane or whether it is due to the geometry of the nerve and labelled axon paths deserves further histological examination. Elucidating the underlying cause of this problem could then direct further development of opsins or promoters [41] with better expression and trafficking characteristics versus development of injection techniques [40,[42][43][44][45] or recombinant viruses [46,47] to increase the efficiency and total number of axons transduced within a nerve.…”

Section: Considerations For Chronic Optical Stimulationmentioning

confidence: 99%

“…A better fundamental understanding of these uptake and transport processes could inform the design of viral vectors for peripheral motor gene therapies. An alternative approach recently taken by several groups is "directed evolution" or high-throughput screening and selection of recombinant AAV variants for a desired trait [46,49,50]. Similarly, the hSyn promoter has been commonly used for peripheral nerve transduction due to its specificity for neural tissues yet relatively strong expression.…”

Section: Virus Delivery Approachesmentioning

confidence: 99%

Viral-mediated optogenetic stimulation of peripheral motor nerves in non-human primates

Williams

Vazquez

Schwartz

2018

Preprint

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Abstract:Objective: Reanimation of muscles paralyzed by disease states such as spinal cord injury remains a much sought after therapeutic goal of neuroprosthetic research. Optogenetic stimulation of peripheral motor nerves expressing light-sensitive opsins is a promising approach to muscle reanimation that may overcome several drawbacks of traditional methods such as functional electrical stimulation (FES). However, the utility of these methods has only been demonstrated in rodents to date, while translation to clinical practice will likely first require demonstration and refinement of these gene therapy techniques in non-human primates.Approach: Two rhesus macaques were injected intramuscularly with either one or both of two optogenetic constructs (AAV6-hSyn-ChR2-eYFP and/or AAV6-hSyn-Chronos-eYFP) to transduce opsin expression in the corresponding nerves. Neuromuscular junctions were targeted for virus delivery using an electrical stimulating injection technique. Functional opsin expression was periodically evaluated up to 13 weeks post-injection by optically stimulating targeted nerves with a 472 nm fiber-coupled laser while recording electromyographic (EMG) responses.Main Results: One monkey demonstrated functional expression of ChR2 at 8 weeks post-injection in each of two injected muscles, while the other monkey briefly exhibited contractions coupled to optical stimulation in a muscle injected with the Chronos construct at 10 weeks. EMG responses to optical stimulation of ChR2-transduced nerves demonstrated graded recruitment relative to both stimulus pulse-width and light intensity, and were able to track stimulus trains up to 16 Hz. In addition, the EMG response to prolonged stimulation showed delayed fatigue over several minutes.Significance: These results demonstrate the feasibility of viral transduction of peripheral motor nerves for functional optical stimulation of motor activity in non-human primates. Subsequently, they represent an important step in translating these optogenetic techniques as a clinically viable gene therapy.

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A Designer AAV Variant Permits Efficient Retrograde Access to Projection Neurons

Cited by 1,137 publications

References 67 publications

Activation of GABAergic Neurons in the Nucleus Accumbens Mediates the Expression of Cocaine-Associated Memory

Activation of GABAergic Neurons in the Nucleus Accumbens Mediates the Expression of Cocaine-Associated Memory

Widespread and targeted gene expression by systemic AAV vectors: Production, purification, and administration

Viral-mediated optogenetic stimulation of peripheral motor nerves in non-human primates

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