A New Promoter Allows Optogenetic Vision Restoration with Enhanced Sensitivity in Macaque Retina

Chaffiol, Antoine; Caplette, Romaine; Jaillard, C.; Brazhnikova, Elena; Desrosiers, Mélissa; Dubus, Élisabeth; Duhamel, Laëtitia; Macé, Émilie; Marre, Olivier; Benoit, Patrick; Hantraye, Philippe; Bemelmans, Alexis-Pierre; Bamberg, Ernst; Duebel, Jens; Sahel, José‐Alain; Picaud, Serge; Dalkara, Deniz

doi:10.1016/j.ymthe.2017.07.011

Cited by 132 publications

(154 citation statements)

References 53 publications

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“…To construct a full model of the reactivated retina we assumed that ganglion cells were placed on a squared grid, with a density equal to the density of transfected cells in the experiment. In a previous study, we found that around 40% of ganglion cells were transfected in the macaque foveal ring (measured from confocal imaging in Chaffiol et al (2017)), and the density of ganglion cells in the macaque fovea has been estimated to 51,108 cells/mm 2 (Ahmed et al, 2003). Each neuron was simulated with an identical LN model, with the parameters (STA, non-linearity) chosen to be equal to the average parameters found in the experimental data (see methods).…”

Section: Acuity Estimation Of the Reactivated Retinamentioning

confidence: 99%

“…We targeted retinal ganglion cells (RGCs) of blind rd1 mice (4/5 weeks old) with an AAV2 encoding ReaChR-mCitrine (a variant of Channel Rhodospin with red-shifted sensitivity) under a pan-neuronal hSyn promoter via intravitreal injections. Details of the gene delivery and optogenetic protein expression has been detailed elsewhere (Sengupta et al, 2016;Chaffiol et al, 2017). Retinas were harvested after 4 weeks for multi-electrode array (MEA) recordings.…”

Section: Optogenetically Activated Ganglion Cells Have Localized Recementioning

confidence: 99%

“…We then tested if the same approach could be applied in the non-human primate retina. We used data from a macaque retina where ganglion cells were transfected with the optogenetic protein CatCh (a variant of Channel-rhodopsin) (Chaffiol et al, 2017). Here AAV2 vectors were produced with a promoter driving the expression of the CatCh protein in ganglion cells.…”

Section: Optogenetically Activated Ganglion Cells Have Localized Recementioning

confidence: 99%

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Optogenetic vision restoration with high resolution

Deny

Sengupta

Caplette

et al. 2018

Preprint

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The majority of inherited retinal degenerations are due to photoreceptor cell death. In many cases ganglion cells are spared making it possible to stimulate them to restore visual function. Several studies (Bi et al., 2006;Lin et al., 2008;Sengupta et al., 2016;Caporale et al., 2011;Berry et al., 2017) have shown that it is possible to express an optogenetic protein in ganglion cells and make them light sensitive. This is a promising strategy to restore vision since optical targeting may be more precise than electrical stimulation with a retinal prothesis. However the spatial resolution of optogenetically-reactivated retinas has not been measured with fine-grained stimulation patterns. Since the optogenetic protein is also expressed in axons, it is unclear if these neurons will only be sensitive to the stimulation of a small region covering their somas and dendrites, or if they will also respond to any stimulation overlapping with their axon, dramatically impairing spatial resolution. Here we recorded responses of mouse and macaque retinas to random checkerboard patterns following an in vivo optogenetic therapy. We show that optogenetically activated ganglion cells are each sensitive to a small region of visual space. A simple model based on this small receptive field predicted accurately their responses to complex stimuli. From this model, we simulated how the entire population of light sensitive ganglion cells would respond to letters of different sizes. We then estimated the maximal acuity expected by a patient, assuming it could make an optimal use of the information delivered by this reactivated retina. The obtained acuity is above the limit of legal blindness. This high spatial resolution is a promising result for future clinical studies.

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Section: Acuity Estimation Of the Reactivated Retinamentioning

confidence: 99%

Section: Optogenetically Activated Ganglion Cells Have Localized Recementioning

confidence: 99%

Section: Optogenetically Activated Ganglion Cells Have Localized Recementioning

confidence: 99%

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Optogenetic vision restoration with high resolution

Deny

Sengupta

Caplette

et al. 2018

Preprint

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“…For all experiments we used GCaMP6s (Chen et al, 2013) under the SNCG promoter (Chaffiol et al, 2017) to specifically target ganglion cells and we used AAV2 as viral vector. To express CoChR (Klapoetke et al, 2014;Shemesh et al, 2017), we used a recently published promoter (In4s-In3-200En-mGluR500P) (Lu et al, 2016), which has been proved to allow specific expression of optogenetic proteins in RBCs.…”

Section: Aav Production and Injectionsmentioning

confidence: 99%

All-optical interrogation of a direction selective retinal circuit by holographic wave front shaping

Spampinato

Ronzitti

Zampini

et al. 2019

Preprint

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Direction selective (DS) ganglion cells (GC) in the retina maintain their tuning across a broad range of light levels. Yet very different circuits can shape their responses from bright to dim light, and their respective contributions are difficult to tease apart. In particular, the contribution of the rod bipolar cell (RBC) primary pathway, a key player in dim light, is unclear. To understand its contribution to DSGC response, we designed an all-optical approach allowing precise manipulation of single retinal neurons. Our system activates single cells in the bipolar cell (BC) layer by two-photon (2P) temporally focused holographic illumination, while recording the activity in the ganglion cell layer by 2P Ca 2 imaging. By doing so, we demonstrate that RBCs provide an asymmetric input to DSGCs, suggesting they contribute to their direction selectivity. Our results suggest that every circuit providing an input to direction selective cells can generate direction selectivity by itself. This hints at a general principle to achieve robust selectivity in sensory areas.A major goal in neuroscience is to understand the circuits that embody the computations performed by sensory neurons. In particular, a striking feature of sensory processing is the ability of neurons to perform the same computation in different contexts. For example, neurons in the piriform cortex represent odor identity while remaining invariant to its exact concentration (Bolding and Franks, 2018). Similarly, neurons in the visual cortex can keep the same orientation tuning curve over different contrasts (Sclar and Freeman, 1982).In the retina, direction selective (DS) ganglion cells (GC) respond selectively to a motion direction in different contexts such as different backgrounds (Chen et al., 2016), different natural scenes (Im and Fried, 2016), and over a broad range of luminosities (Pearson and Kerschensteiner, 2015; Vaney et al., 2001;Yao et al., 2018). Switching the average luminance from dim to bright light will change the dominant circuits that convey visual information inside the retina, leading to major changes in the responses of ganglion cells (Pearson and Kerschensteiner, 2015;Wässle, 2004). For example, cells will change their ON-OFF polarity (Tikidji-Hamburyan et al., 2015). It is therefore particularly striking that direction selectivity remains constant across the ten billion fold change of luminance separating day from night (Yao et al., 2018), despite the involvement of distinct circuits. A major challenge is to understand how this feature is achieved.During daylight conditions, where rods are saturated and cones transmit light input, the major component responsible for direction selectivity is a functional asymmetry. This results either from an asymmetric inhibition or an asymmetric morphology. For example, in the ON-OFF

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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%

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 New Promoter Allows Optogenetic Vision Restoration with Enhanced Sensitivity in Macaque Retina

Cited by 132 publications

References 53 publications

Optogenetic vision restoration with high resolution

Optogenetic vision restoration with high resolution

All-optical interrogation of a direction selective retinal circuit by holographic wave front shaping

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

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