Primate Retina Research Articles

Intrinsically-photosensitive ganglion cells in the primate retina: Anatomy, physiology and behavioral roles

Intrinsically-photosensitive ganglion cells in the primate retina: Anatomy, physiology and behavioral roles

A synaptic signature for ON- and OFF-center parasol ganglion cells of the primate retina.

In the primate retina, parasol ganglion cells contribute to the primary visual pathway via the magnocellular division of the lateral geniculate nucleus, display ON and OFF concentric receptive field structure, nonlinear spatial summation, and high achromatic temporal-contrast sensitivity. Parasol cells may be homologous to the alpha-Y cells of nonprimate mammals where evidence suggests that N-methyl-D-aspartate (NMDA) receptor-mediated synaptic excitation as well as glycinergic disinhibition play critical roles in contrast sensitivity, acting asymmetrically in OFF- but not ON-pathways. Here, light-evoked synaptic currents were recorded in the macaque monkey retina in vitro to examine the circuitry underlying parasol cell receptive field properties. Synaptic excitation in both ON and OFF types was mediated by NMDA as well as α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)/kainate glutamate receptors. The NMDA-mediated current-voltage relationship suggested high Mg2+ affinity such that at physiological potentials, NMDA receptors contributed ∼20% of the total excitatory conductance evoked by moderate stimulus contrasts and temporal frequencies. Postsynaptic inhibition in both ON and OFF cells was dominated by a large glycinergic "crossover" conductance, with a relatively small contribution from GABAergic feedforward inhibition. However, crossover inhibition was largely rectified, greatly diminished at low stimulus contrasts, and did not contribute, via disinhibition, to contrast sensitivity. In addition, attenuation of GABAergic and glycinergic synaptic inhibition left center-surround and Y-type receptive field structure and high temporal sensitivity fundamentally intact and clearly derived from modulation of excitatory bipolar cell output. Thus, the characteristic spatial and temporal-contrast sensitivity of the primate parasol cell arises presynaptically and is governed primarily by modulation of the large AMPA/kainate receptor-mediated excitatory conductance. Moreover, the negative feedback responsible for the receptive field surround must derive from a nonGABAergic mechanism.

Rod Photoreceptors Express GPR55 in the Adult Vervet Monkey Retina

Cannabinoids exert their actions mainly through two receptors, the cannabinoid CB1 receptor (CB1R) and cannabinoid CB2 receptor (CB2R). In recent years, the G-protein coupled receptor 55 (GPR55) was suggested as a cannabinoid receptor based on its activation by anandamide and tetrahydrocannabinol. Yet, its formal classification is still a matter of debate. CB1R and CB2R expression patterns are well described for rodent and monkey retinas. In the monkey retina, CB1R has been localized in its neural (cone photoreceptor, horizontal, bipolar, amacrine and ganglion cells) and CB2R in glial components (Müller cells). The aim of this study was to determine the expression pattern of GPR55 in the monkey retina by using confocal microscopy. Our results show that GPR55 is strictly localized in the photoreceptor layer of the extrafoveal portion of the retina. Co-immunolabeling of GPR55 with rhodopsin, the photosensitive pigment in rods, revealed a clear overlap of expression throughout the rod structure with most prominent staining in the inner segments. Additionally, double-label of GPR55 with calbindin, a specific marker for cone photoreceptors in the primate retina, allowed us to exclude expression of GPR55 in cones. The labeling of GPR55 in rods was further assessed with a 3D visualization in the XZ and YZ planes thus confirming its exclusive expression in rods. These results provide data on the distribution of GPR55 in the monkey retina, different than CB1R and CB2R. The presence of GPR55 in rods suggests a function of this receptor in scotopic vision that needs to be demonstrated.

Inner retinal inhibition shapes the receptive field of retinal ganglion cells in primate

The centre–surround organisation of receptive fields is a feature of most retinal ganglion cells (RGCs) and is critical for spatial discrimination and contrast detection. Although lateral inhibitory processes are known to be important in generating the receptive field surround, the contribution of each of the two synaptic layers in the primate retina remains unclear. Here we studied the spatial organisation of excitatory and inhibitory synaptic inputs onto ON and OFF ganglion cells in the primate retina. All RGCs showed an increase in excitation in response to stimulus of preferred polarity. Inhibition onto RGCs comprised two types of responses to preferred polarity: some RGCs showed an increase in inhibition whilst others showed removal of tonic inhibition. Excitatory inputs were strongly spatially tuned but inhibitory inputs showed more variable organisation: in some neurons they were as strongly tuned as excitation, and in others inhibitory inputs showed no spatial tuning. We targeted one source of inner retinal inhibition by functionally ablating spiking amacrine cells with bath application of tetrodotoxin (TTX). TTX significantly reduced the spatial tuning of excitatory inputs. In addition, TTX reduced inhibition onto those RGCs where a stimulus of preferred polarity increased inhibition. Reconstruction of the spatial tuning properties by somatic injection of excitatory and inhibitory synaptic conductances verified that TTX-mediated inhibition onto bipolar cells increases the strength of the surround in RGC spiking output. These results indicate that in the primate retina inhibitory mechanisms in the inner plexiform layer sharpen the spatial tuning of ganglion cells.

NaV1.1 Channels in Axon Initial Segments of Bipolar Cells Augment Input to Magnocellular Visual Pathways in the Primate Retina

In the primate visual system, the ganglion cells of the magnocellular pathway underlie motion and flicker detection and are relatively transient, while the more sustained ganglion cells of the parvocellular pathway have comparatively lower temporal resolution, but encode higher spatial frequencies. Although it is presumed that functional differences in bipolar cells contribute to the tuning of the two pathways, the properties of the relevant bipolar cells have not yet been examined in detail. Here, by making patch-clamp recordings in acute slices of macaque retina, we show that the bipolar cells within the magnocellular pathway, but not the parvocellular pathway, exhibit voltage-gated sodium (NaV), T-type calcium (CaV), and hyperpolarization-activated, cyclic nucleotide-gated (HCN) currents, and can generate action potentials. Using immunohistochemistry in macaque and human retinae, we show that NaV1.1 is concentrated in an axon initial segment (AIS)-like region of magnocellular pathway bipolar cells, a specialization not seen in transient bipolar cells of other vertebrates. In contrast, CaV3.1 channels were localized to the somatodendritic compartment and proximal axon, but were excluded from the AIS, while HCN1 channels were concentrated in the axon terminal boutons. Simulations using a compartmental model reproduced physiological results and indicate that magnocellular pathway bipolar cells initiate spikes in the AIS. Finally, we demonstrate that NaV channels in bipolar cells augment excitatory input to parasol ganglion cells of the magnocellular pathway. Overall, the results demonstrate that selective expression of voltage-gated channels contributes to the establishment of parallel processing in the major visual pathways of the primate retina.

Processing of S-cone signals in the inner plexiform layer of the mammalian retina

Color information is encoded by two parallel pathways in the mammalian retina. One pathway compares signals from long- and middle-wavelength sensitive cones and generates red-green opponency. The other compares signals from short- and middle-/long-wavelength sensitive cones and generates blue-green (yellow) opponency. Whereas both pathways operate in trichromatic primates (including humans), the fundamental, phylogenetically ancient color mechanism shared among most mammals is blue-green opponency. In this review, we summarize the current understanding of how signals from short-wavelength sensitive cones are processed in the primate and nonprimate mammalian retina, with a focus on the inner plexiform layer where bipolar, amacrine, and ganglion cell processes interact to facilitate the generation of blue-green opponency.

Amino acid immunoreactivity in normal human retina and after brachytherapy

We localised amino acids in the mid‐peripheral aged human retina and a retina that had undergone radiation treatment 10 years earlier. The distribution pattern of glutamate, γ‐amino butyric acid (GABA), glycine, glutamine and taurine, reflected patterns established in the primate retina. The retina that had undergone radiation exposure displayed both anatomical and neurochemical remodelling. The proximal retina comprised around 40 to 45 per cent of the total retina and neuronal kinesis and aberrant neuronal projections were also present. Amino acid neurochemistry was strikingly different with Müller cells displaying GABA loading, glycinergic neurons displaced and displaying a very high level of glycine labelling. We conclude that radiation exposure triggered these changes in the human retina and likely reflects general remodelling of structure and function following ischaemic damage to endothelial cells.

The Effect of Lutein Supplementation on Blood Plasma Levels of Complement Factor D, C5a and C3d

Lutein is selectively taken up by the primate retina and plays an important role as a filter for harmful blue light and as an antioxidant. Recent studies have shown that lutein has systemic anti-inflammatory properties. Dietary lutein has been associated with reduced circulating levels of inflammatory biomarkers such as CRP and sICAM. Whether lutein also affects activation of the complement system has not yet been addressed and was the purpose of the study described here. Seventy-two subjects with signs of early macular degeneration were randomly assigned to receive either a 10 mg lutein supplement or a placebo during one year. EDTA blood samples were collected at 0, 4, 8 and 12 months. Complement factor D (CFD), a rate limiting component of the alternative pathway of complement activation and the complement activation products C5a and C3d were determined in the plasma samples by ELISA. A significant 0.11 µg/ml monthly decrease in plasma CFD concentration was observed in the lutein group (p<0.001), resulting in a 51% decrease from 2.3 µg/ml at baseline to 1.0 µg/ml at 12 months. The C5a concentration showed a significant 0.063ng/ml monthly decrease in the lutein group (p<0.001) resulting in a 36% decrease from 2.2ng/ml at baseline to 1.6ng/ml at 12 months. The C3d concentration showed a significant 0.19µg/ml monthly decrease in the lutein group (p=0.004) that gave rise to a 9% decrease from 15.4µg/ml at baseline to 14.4µg/ml at 12 months. In the placebo group we found a significant 0.04 µg/ml monthly decrease in plasma CFD concentration, whereas no changes were observed for C5a and C3d. Lutein supplementation markedly decreases circulating levels of the complement factors CFD, C5a and C3d levels, which might allow a simple method to control this inflammatory pathway of the innate immune system.

Correction: AAV9 Targets Cone Photoreceptors in the Nonhuman Primate Retina

A project funding the seventh author was incorrectly omitted from the Funding Statement. The second sentence of the Funding Statement should read: This work was also supported by the Foundation Fighting Blindness (LHV, JB), its CHOP-PENN Pediatric Center for Retinal Degenerations (JB), Transdisciplinary Award Program in Translational Medicine and Therapeutics (TAPITMAT) from the University of Pennsylvania (LHV, JB), Research to Prevent Blindness, the Paul and Evanina Mackall Foundation Trust at Scheie Eye Institute, the F. M. Kirby Foundation (JB) and Treatrush, European Commission (JB).

Distinct synaptic mechanisms create parallel S-ON and S-OFF color opponent pathways in the primate retina

Anatomical and physiological approaches are beginning to reveal the synaptic origins of parallel ON- and OFF-pathway retinal circuits for the transmission of short (S-) wavelength sensitive cone signals in the primate retina. Anatomical data suggest that synaptic output from S-cones is largely segregated; central elements of synaptic triads arise almost exclusively from the "blue-cone" bipolar cell, a presumed ON bipolar, whereas triad-associated contacts derive primarily from the "flat" midget bipolar cell, a hyperpolarizing, OFF bipolar. Similarly, horizontal cell connectivity is also segregated, with only the H2 cell-type receiving numerous contacts from S-cones. Negative feedback from long (L-) and middle (M-) wavelength sensitive cones via the H2 horizontal cells elicits an antagonistic surround in S-cones demonstrating that S versus L + M or "blue-yellow" opponency is first established in the S-cone. However, the S-cone output utilizes distinct synaptic mechanisms to create color opponency at the ganglion cell level. The blue-cone bipolar cell is presynaptic to the small bistratified, "blue-ON" ganglion cell. S versus L + M cone opponency arises postsynaptically by converging S-ON and LM-OFF excitatory bipolar inputs to the ganglion cell's bistratified dendritic tree. The common L + M cone surrounds of the parallel S-ON and LM-OFF cone bipolar inputs appear to cancel resulting in "blue-yellow" antagonism without center-surround spatial opponency. By contrast, in midget ganglion cells, opponency arises by the differential weighting of cone inputs to the receptive field center versus surround. In the macula, the "private-line" connection from a midget ganglion cell to a single cone predicts that S versus L + M opponency is transmitted from the S-cone to the S-OFF midget bipolar and ganglion cell. Beyond the macula, OFF-midget ganglion cell dendritic trees enlarge and collect additional input from multiple L and M cones. Thus S-OFF opponency via the midget pathway would be expected to become more complex in the near retinal periphery as L and/or M and S cone inputs sum to the receptive field center. An important goal for further investigation will be to explore the hypothesis that distinct bistratified S-ON versus midget S-OFF retinal circuits are the substrates for human psychophysical detection mechanisms attributed to S-ON versus S-OFF perceptual channels.

Fast inference in generalized linear models via expected log-likelihoods

Generalized linear models play an essential role in a wide variety of statistical applications. This paper discusses an approximation of the likelihood in these models that can greatly facilitate computation. The basic idea is to replace a sum that appears in the exact log-likelihood by an expectation over the model covariates; the resulting "expected log-likelihood" can in many cases be computed significantly faster than the exact log-likelihood. In many neuroscience experiments the distribution over model covariates is controlled by the experimenter and the expected log-likelihood approximation becomes particularly useful; for example, estimators based on maximizing this expected log-likelihood (or a penalized version thereof) can often be obtained with orders of magnitude computational savings compared to the exact maximum likelihood estimators. A risk analysis establishes that these maximum EL estimators often come with little cost in accuracy (and in some cases even improved accuracy) compared to standard maximum likelihood estimates. Finally, we find that these methods can significantly decrease the computation time of marginal likelihood calculations for model selection and of Markov chain Monte Carlo methods for sampling from the posterior parameter distribution. We illustrate our results by applying these methods to a computationally-challenging dataset of neural spike trains obtained via large-scale multi-electrode recordings in the primate retina.

Latency and rate coding in a spiking model of retina

We describe a family of spiking retina computational models for large-scale simulations of the visual pathways. The models reproduce the overall spatial, temporal, and chromatic structure of the receptive fields of midget and parasol retinal ganglion cells (RGCs) of the primate retina, as well as their contrast response, while using fairly simple models of bipolar, horizontal, amacrine, and RGCs. These retina models provide input to a realistic model of visual cortex, presented in a separate submission. The retina model simulates cone and bipolar cell dynamics using a damped wave equation. Parameters are so chosen that an impulse retinal input gives rise to a damped biphasic output current, and that a coherent stimulus elicits a substantially stronger response than an incoherent stimulus of the same power (Figure ​(Figure1).1). Static or low-frequency visual inputs are smoothly rejected. In contrast to biology, this stage of the model is linear; rectification and adaptation mechanisms are all realized in the spiking ganglion cell layer. In the ganglion cell layer parasol, midget, and SBC cells are simulated. The spatio-temporal receptive fields of different RGCs are not prewired; they emerge via the lateral interactions between the horizontal, bipolar, and amacrine cells. Spike generation in the RGCs is modeled using spiking neurons with two dynamic variables and an intrinsic adaptation mechanism. Midget RGCs have an additional leaky temporal integration of the input current, to account for the relatively tonic character of their responses. Parasol RGCs have an additional partial divisive normalization of the input current, to reproduce the saturating parasol response to relative contrast of the stimulus. Simplified amacrine-cell dynamics is introduced to provide for the spatiotemporal structure of the parasol receptive field surrounds. Figure 1 A-E: model response to coherent vs. randomized stimulus. Moving edge stimulus is presented (A) and elicits strong response (B). Same stimulus with fixed random permutation of pixels (C) elicits much weaker response (D). (E) compares the total responses ... Desired latency encoding of input features is achieved by calibrating the parameters of the spiking RGC conductance dynamics. A near logarithmic latency to the first spike, for a step up in the input signal, provides for a good contrast invariance of the relative spike timing in the response. A combination of spike-latency and spike-count encoding of the input stimulus, together with the enhanced response of the simulated retina to coherent stimuli, results in an informative yet not too noisy spiking input to the LGN and V1; the information content analysis of the RGC spike-train vocabulary will be presented. This retina model was used to produce a fully-emergent orientation tuning in a spiking model [1] of V1 cortex.

Spontaneous emergence of simple and complex receptive fields in a spiking model of V1

Brain Corporation is engaged in a multi-year project to build a spiking model of vision, paying special attention to the anatomy and physiology of the mammalian visual system. While it is relatively easy to hand-tune V1 to get simple and complex cells, it is not clear how to arrange connectivity in other cortical areas to get appropriate receptive fields, or what the appropriate receptive fields even should be. Instead of pre-wiring cortical connectivity according to a computational theory of how vision should work, we start with a generic tabula rasa spiking network having multiple cortical layers and neuronal types (single-compartment RS, FS, LTS cells). The goal is to find the anatomical and physiological parameters so that the appropriate connectivity emerges through STDP and visual experience. Since we know exactly what kind of receptive fields and visual responses are in V1, we build a smaller model of retina-LGN-V1 pathway and tune the STDP parameters so that the expected responses emerge. Admittedly, there are many free parameters that could be tuned to get V1-like responses; our choice is motivated by in-vitro recordings and other published data, whenever possible. Once we trust what we see in V1, we are ready to copy and paste the cortical model to implement V2, V3, V4, and IT areas with the hope that useful connectivity, receptive fields, and visual responses emerge. Our large-scale simulations of the spiking model of the visual system show spontaneous emergence of simple and complex cells, orientation domains, end-stopping receptive fields, extra-classical receptive fields with tuned surround suppression, color opponency that depends on the eccentricity of the receptive field, contrast invariance, and many other features that are routinely recorded in V1. Since the visual model exhibits micro- and full saccades, we observe perceptual behavior, such as the emergence of bottom-up (pop-out) attention. The model underscores the importance of spike-timing dynamics, inhibition, saccadic mechanism, and it imposes important restrictions on the possible types of STDP to model early visual processing. This presentation is meant to be a flagship introduction to the whole effort by Brain Corporation to build spiking model of vision. There are multiple other submissions elaborating details of our modeling effort. The figure summarizes the gross anatomy of the spiking model of the visual system, including area MT, Superior Colliculus (SC), Frontal Pursuit Area (FPA), Flocculus (Floc), and Brainstem (B.Stem). The retina covers 10 by 10 degrees of visual field with the density of L, M, and S cones and receptive field sizes of Midget, Parasol, and SBC retino-ganglion cells corresponding to 4 degrees of eccentricity of a primate retina (collaboration with EJ Chichilnisky lab at Salk Institute). The model consists of more than 1M single-compartment neurons of RS, FS, and LTS types (Izhikevich, 2007 Dynamical Systems in Neuroscience) and 0.25B synapses having AMPA, NMDA, GABA_A and GABA_B conductances, axonal conduction delays, and STDP. In this presentation, we summarize these and other anatomical and physiological assumptions, describe major results and point to other submissions by Brain Corporation scientists for more details. Figure 1

Müller cells express the cannabinoid CB2 receptor in the vervet monkey retina

AbstractThe presence of the cannabinoid receptor type 1 (CB1R) has been largely documented in the rodent and primate retinae in recent years. There is, however, some controversy concerning the presence of the CB2 receptor (CB2R) within the central nervous system. Only recently, CB2R has been found in the rodent retina, but its presence in the primate retina has not yet been demonstrated. The aim of this study was twofold: 1) to characterize the distribution patterns of CB2R in the monkey retina and compare this distribution with that previously reported for CB1R and 2) to resolve the controversy on the presence of CB2R in the neural component of the retina. We therefore thoroughly examined the cellular localization of CB2R in the vervet monkey (Chlorocebus sabeus) retina, using confocal microscopy. Our results demonstrate that CB2R, like CB1R, is present throughout the retinal layers, but with striking dissimilarities. Double labeling of CB2R and glutamine synthetase shows that CB2R is restricted to Müller cell processes, extending from the internal limiting membrane, with very low staining, to the external limiting membrane, with heavy labeling. We conclude that CB2R is indeed present in the retina but exclusively in the retinal glia, whereas CB1R is expressed only in the neuroretina. These results extend our knowledge on the expression and distribution of cannabinoid receptors in the monkey retina, although further experiments are still needed to clarify their role in retinal functions. J. Comp. Neurol. J. Comp. Neurol. 521:2399–2415, 2013. © 2013 Wiley Periodicals, Inc.

Müller cells express the cannabinoid CB2 receptor in the vervet monkey retina

The presence of the cannabinoid receptor type 1 (CB1R) has been largely documented in the rodent and primate retinae in recent years. There is, however, some controversy concerning the presence of the CB2 receptor (CB2R) within the central nervous system. Only recently, CB2R has been found in the rodent retina, but its presence in the primate retina has not yet been demonstrated. The aim of this study was twofold: 1) to characterize the distribution patterns of CB2R in the monkey retina and compare this distribution with that previously reported for CB1R and 2) to resolve the controversy on the presence of CB2R in the neural component of the retina. We therefore thoroughly examined the cellular localization of CB2R in the vervet monkey (Chlorocebus sabeus) retina, using confocal microscopy. Our results demonstrate that CB2R, like CB1R, is present throughout the retinal layers, but with striking dissimilarities. Double labeling of CB2R and glutamine synthetase shows that CB2R is restricted to Müller cell processes, extending from the internal limiting membrane, with very low staining, to the external limiting membrane, with heavy labeling. We conclude that CB2R is indeed present in the retina but exclusively in the retinal glia, whereas CB1R is expressed only in the neuroretina. These results extend our knowledge on the expression and distribution of cannabinoid receptors in the monkey retina, although further experiments are still needed to clarify their role in retinal functions.

Magnetic resonance imaging of the retina: from mice to men.

This mini-review provides an overview of magnetic resonance imaging (MRI) applications to study rodent, cat, non-human primate, and human retinas. These techniques include T(1) - and T(2) -weighted anatomical, diffusion, blood flow, blood volume, blood-oxygenation level dependent, manganese-enhanced, physiological, and functional MRI. Applications to study the retinas in diabetic retinopathy, glaucoma, and retinal degeneration are also reviewed. MRI offers some unique advantages compared with existing imaging techniques and has the potential to further our understanding of physiology and function in healthy and diseased retinas.

Controlling gain one photon at a time

Adaptation is a salient property of sensory processing. All adaptational or gain control mechanisms face the challenge of obtaining a reliable estimate of the property of the input to be adapted to and obtaining this estimate sufficiently rapidly to be useful. Here, we explore how the primate retina balances the need to change gain rapidly and reliably when photons arrive rarely at individual rod photoreceptors. We find that the weakest backgrounds that decrease the gain of the retinal output signals are similar to those that increase human behavioral threshold, and identify a novel site of gain control in the retinal circuitry. Thus, surprisingly, the gain of retinal signals begins to decrease essentially as soon as background lights are detectable; under these conditions, gain control does not rely on a highly averaged estimate of the photon count, but instead signals from individual photon absorptions trigger changes in gain. DOI:http://dx.doi.org/10.7554/eLife.00467.001.

A model-based spike sorting algorithm for removing correlation artifacts in multi-neuron recordings.

We examine the problem of estimating the spike trains of multiple neurons from voltage traces recorded on one or more extracellular electrodes. Traditional spike-sorting methods rely on thresholding or clustering of recorded signals to identify spikes. While these methods can detect a large fraction of the spikes from a recording, they generally fail to identify synchronous or near-synchronous spikes: cases in which multiple spikes overlap. Here we investigate the geometry of failures in traditional sorting algorithms, and document the prevalence of such errors in multi-electrode recordings from primate retina. We then develop a method for multi-neuron spike sorting using a model that explicitly accounts for the superposition of spike waveforms. We model the recorded voltage traces as a linear combination of spike waveforms plus a stochastic background component of correlated Gaussian noise. Combining this measurement model with a Bernoulli prior over binary spike trains yields a posterior distribution for spikes given the recorded data. We introduce a greedy algorithm to maximize this posterior that we call “binary pursuit”. The algorithm allows modest variability in spike waveforms and recovers spike times with higher precision than the voltage sampling rate. This method substantially corrects cross-correlation artifacts that arise with conventional methods, and substantially outperforms clustering methods on both real and simulated data. Finally, we develop diagnostic tools that can be used to assess errors in spike sorting in the absence of ground truth.

Focal Electrical Stimulation of Major Ganglion Cell Types in the Primate Retina for the Design of Visual Prostheses

Electrical stimulation of retinal neurons with an advanced retinal prosthesis may eventually provide high-resolution artificial vision to the blind. However, the success of future prostheses depends on the ability to activate the major parallel visual pathways of the human visual system. Electrical stimulation of the five numerically dominant retinal ganglion cell types was investigated by simultaneous stimulation and recording in isolated peripheral primate (Macaca sp.) retina using multi-electrode arrays. ON and OFF midget, ON and OFF parasol, and small bistratified ganglion cells could all be activated directly to fire a single spike with submillisecond latency using brief pulses of current within established safety limits. Thresholds for electrical stimulation were similar in all five cell types. In many cases, a single cell could be specifically activated without activating neighboring cells of the same type or other types. These findings support the feasibility of direct electrical stimulation of the major visual pathways at or near their native spatial and temporal resolution.

Assessment of Tropism and Effectiveness of New Primate-Derived Hybrid Recombinant AAV Serotypes in the Mouse and Primate Retina

Adeno-associated viral vectors (AAV) have been shown to be safe in the treatment of retinal degenerations in clinical trials. Thus, improving the efficiency of viral gene delivery has become increasingly important to increase the success of clinical trials. In this study, structural domains of different rAAV serotypes isolated from primate brain were combined to create novel hybrid recombinant AAV serotypes, rAAV2/rec2 and rAAV2/rec3. The efficacy of these novel serotypes were assessed in wild type mice and in two models of retinal degeneration (the Abca4−/− mouse which is a model for Stargardt disease and in the Pde6brd1/rd1 mouse) in vivo, in primate tissue ex-vivo, and in the human-derived SH-SY5Y cell line, using an identical AAV2 expression cassette. We show that these novel hybrid serotypes can transduce retinal tissue in mice and primates efficiently, although no more than AAV2/2 and rAAV2/5 serotypes. Transduction efficiency appeared lower in the Abca4−/− mouse compared to wild type with all vectors tested, suggesting an effect of specific retinal diseases on the efficiency of gene delivery. Shuffling of AAV capsid domains may have clinical applications for patients who develop T-cell immune responses following AAV gene therapy, as specific peptide antigen sequences could be substituted using this technique prior to vector re-treatments.