Comparison of Retinal Ganglion Cell Responses to Different Voltage Stimulation Parameters in Normal and rd1 Mouse Retina / 의학물리

Retinal prostheses are being developed to restore vision for the blind with retinal diseases such as retinitis pigmentosa (RP) or age-related macular degeneration (AMD). Since retinal prostheses depend upon electrical stimulation to control neural activity, optimal stimulation parameters for successful encoding of visual information are one of the most important requirements to enable visual perception. Therefore, in this paper, we focused on retinal ganglion cell (RGC) responses to different voltage stimulation parameters and compared threshold charge densities in normal and rd1 mice. For this purpose, we used in vitro preparation for the retina of normal and rd1 mice on micro-electrode arrays. When the neural network of rd1 mouse retinas is stimulated with voltage-controlled pulses, RGCs in degenerated retina also respond to voltage amplitude or voltage duration modulation as well in wild-type RGCs. But the temporal pattern of RGCs response is very different; in wild-type RGCs, single peak within 100 ms appears while in RGCs in degenerated retina multiple peaks (~4 peaks) with ~10 Hz rhythm within 400 ms appear. The thresholds for electrical activation of RGCs are overall more elevated in rd1 mouse retinas compared to wild-type mouse retinas: The thresholds for activation of RGCs in rd1 mouse retinas were on average two times higher (70.50~99.87micronC/cm2 vs. 37.23~61.65micronC/cm2) in the experiment of voltage amplitude modulation and five times higher (120.5~170.6micronC/cm2 vs. 22.69~37.57micronC/cm2) in the experiment of voltage duration modulation than those in wild-type mouse retinas. This is compatible with the findings from human studies that the currents required for evoking visual percepts in RP patients is much higher than those needed in healthy individuals. These results will be used as a guideline for optimal stimulation parameters for upcoming Korean-type retinal prosthesis.

Accurate Representation of Light-intensity Information by the Neural Activities of Independently Firing Retinal Ganglion Cells

For successful restoration of visual function by a visual neural prosthesis such as retinal implant, electrical stimulation should evoke neural responses so that the information on visual input is properly represented. A stimulation strategy, which means a method for generating stimulation waveforms based on visual input, should be developed for this purpose. We proposed to use the decoding of visual input from retinal ganglion cell (RGC) responses for the evaluation of stimulus encoding strategy. This is based on the assumption that reliable encoding of visual information in RGC responses is required to enable successful visual perception. The main purpose of this study was to determine the influence of inter-dependence among stimulated RGCs activities on decoding accuracy. Light intensity variations were decoded from multiunit RGC spike trains using an optimal linear filter. More accurate decoding was possible when different types of RGCs were used together as input. Decoding accuracy was enhanced with independently firing RGCs compared to synchronously firing RGCs. This implies that stimulation of independently-firing RGCs and RGCs of different types may be beneficial for visual function restoration by retinal prosthesis.

Electrically-evoked Neural Activities of rd1 Mice Retinal Ganglion Cells by Repetitive Pulse Stimulation

For successful visual perception by visual prosthesis using electrical stimulation, it is essential to develop an effective stimulation strategy based on understanding of retinal ganglion cell (RGC) responses to electrical stimulation. We studied RGC responses to repetitive electrical stimulation pulses to develop a stimulation strategy using stimulation pulse frequency modulation. Retinal patches of photoreceptor-degenerated retinas from rd1 mice were attached to a planar multi-electrode array (MEA) and RGC spike trains responding to electrical stimulation pulse trains with various pulse frequencies were observed. RGC responses were strongly dependent on inter-pulse interval when it was varied from 500 to 10 ms. Although the evoked spikes were suppressed with increasing pulse rate, the number of evoked spikes were >60% of the maximal responses when the inter-pulse intervals exceeded 100 ms. Based on this, we investigated the modulation of evoked RGC firing rates while increasing the pulse frequency from 1 to 10 pulses per second (or Hz) to deduce the optimal pulse frequency range for modulation of RGC response strength. RGC response strength monotonically and linearly increased within the stimulation frequency of 1~9 Hz. The results suggest that the evoked neural activities of RGCs in degenerated retina can be reliably controlled by pulse frequency modulation, and may be used as a stimulation strategy for visual neural prosthesis.

Functional Connectivity Map of Retinal Ganglion Cells for Retinal Prosthesis

Retinal prostheses are being developed to restore vision for the blind with retinal diseases such as retinitis pigmentosa (RP) or age-related macular degeneration (AMD). Among the many issues for prosthesis development, stimulation encoding strategy is one of the most essential electrophysiological issues. The more we understand the retinal circuitry how it encodes and processes visual information, the greater it could help decide stimulation encoding strategy for retinal prosthesis. Therefore, we examined how retinal ganglion cells (RGCs) in in-vitro retinal preparation act together to encode a visual scene with multielectrode array (MEA). Simultaneous recording of many RGCs with MEA showed that nearby neurons often fired synchronously, with spike delays mostly within 1 ms range. This synchronized firing - narrow correlation - was blocked by gap junction blocker, heptanol, but not by glutamatergic synapse blocker, kynurenic acid. By tracking down all the RGC pairs which showed narrow correlation, we could harvest 40 functional connectivity maps of RGCs which showed the cell cluster firing together. We suggest that finding functional connectivity map would be useful in stimulation encoding strategy for the retinal prosthesis since stimulating the cluster of RGCs would be more efficient than separately stimulating each individual RGC.

Comparison of Retinal Waveform between Normal and rd/rd Mouse / 의학물리

Retinal prosthesis is regarded as the most feasible method for the blind caused by retinal diseases such as retinitis pigmentosa or age-related macular degeneration. One of the prerequisites for the success of retinal prosthesis is the optimization of the electrical stimuli applied through the prosthesis. Since electrical characteristics of degenerate retina are expected to differ from those of normal retina, we investigated differences of the retinal waveforms in normal and degenerate retina to provide a guideline for the optimization of electrical stimulation for the upcoming prosthesis. After isolation of retina, retinal patch was attached with the ganglion cell side facing the surface of microelectrode arrays (MEA). 8x8 grid layout MEA (electrode diameter: 30micrometer, electrode spacing: 200micrometer, and impedance: 50 k omega at 1 kHz) was used to record in-vitro retinal ganglion cell activity. In normal mice (C57BL/6J strain) of postnatal day 28, only short duration (<2 ms) retinal spikes were recorded. In rd/rd mice (C3H/HeJ strain), besides normal spikes, waveform with longer duration (~100 ms), the slow wave component was recorded. We attempted to understand the mechanism of this slow wave component in degenerate retina using various synaptic blockers. We suggest that stronger glutamatergic input from bipolar cell to the ganglion cell in rd/rd mouse than normal mouse contributes the most to this slow wave component. Out of many degenerative changes, we favor elimination of the inhibitory horizontal input to bipolar cells as a main contributor for a relatively stronger input from bipolar cell to ganglion cell in rd/rd mouse.

Spatiotemporal Analysis of Retinal Waveform using Independent Component Analysis in Normal and rd/rd Mouse / 의학물리

It is expected that synaptic construction and electrical characteristics in degenerate retina might be different from those in normal retina. Therefore, we analyzed the retinal waveform recorded with multielectrode array in normal and degenerate retina using principal component analysis (PCA) and independent component analysis (ICA) and compared the results. PCA is a well established method for retinal waveform while ICA has not tried for retinal waveform analysis. We programmed ICA toolbox for spatiotemporal analysis of retinal waveform. In normal mouse, the MEA spatial map shows a single hot spot perfectly matched with PCA-derived ON or OFF ganglion cell response. However in rd/rd mouse, the MEA spatial map shows numerous hot and cold spots whose underlying interactions and mechanisms need further investigation for better understanding.

Characteristics of Light-evoked Retinal Ganglion Cell Activity with Postnatal Maturation in SD Rat / 의학물리

As part of Korean retinal prosthesis project, we have provided preliminary experimental results regarding voltage parameters for the stimulation of chemically degenerated rabbit retina. Since our APB-treated chemically degenerated retina is only ON-pathway blocked, now we switch our experiments to more appropriate retinal degeneration model, genetically degenerated retina model (RD mouse: rd/rd (C3H/HeJ)). Before studying with RD mouse, we started control experiments with normal SD rat to understand characteristics of retinal ganglion cell activity with postnatal maturation in rodents. Ganglion cell activities were recorded with 8x8 multi-electrode array. Moving spontaneous bursts appeared until postnatal day of 15. During pre-eye opening period, no light evoked response appeared. After postnatal day of 2 weeks (post-eye opening period), ON-, OFF- and ON/OFF response appeared. The fractional distributions of ON, OFF, and ON/OFF ganglion cell is about 40%, 50%, and 5%. The percentage (%) of light evoked response in each dorso-temporal, ventral, and dorso-nasal area of eye is about 50%, 37.5% and 12.5%, respectively. We concluded that the optimal period for experiment in rodent is about postnatal day of 2~3 weeks.