Implantation of stimulation electrodes in the subretinal space to demonstrate cortical responses in Yucatan minipig in the course of visual prosthesis development.

PURPOSE: During the course of the development of visual prostheses, subretinal stimulation films were implanted in micropigs in order to prove the feasibility of subretinal electrical stimulation with subsequent cortical response. One aim was to demonstrate that epidural recording of visual evoked potentials is possible in the micropig. METHODS: Film-bound stimulation electrode arrays were placed in the subretinal space of micropigs. This enabled the retina to be stimulated subretinally. Since conventional visual evoked potential (VEP) measuring is virtually impossible in the pig from the neurosurgical point of view, epidural recording electrode arrays were positioned over the visual cortex as permanent electrodes. RESULTS: The feasibility of temporary implantation of film-bound stimulation electrode arrays was successfully demonstrated in the micropig model. On stimulation with monopolar voltage pulses (1000 to 3000 mV), reproducible epidural VEP measurements (5 to 10 micronV) were detected. CONCLUSIONS: The feasibility of subretinal stimulation of the retina was demonstrated in a retinal model that is similar to the human retina. This animal model therefore offers a suitable means of studying the tolerability of stimulation situations in the course of visual prosthesis development.

Chick eyes under cycloplegia compensate for spectacle lenses despite six-hydroxy dopamine treatment.

PURPOSE: To test whether eye growth changes produced by spectacle lens wear are mediated by changes in ciliary muscle tonus in chicks. METHODS: Because there is evidence that deprivation myopia is based on a local-retinal mechanism in the eye that probably remains functional after cycloplegia as well as after ciliary ganglion or Edinger-Westphal lesions, none of these treatments provides insight into whether accommodation tonus is also important in the control of axial eye growth. Because 6-hydroxy dopamine (6-OHDA) suppresses deprivation myopia, to isolate growth changes mediated by accommodation the authors injected 6-OHDA and paralyzed accommodation in addition (by corneal application of vecuroniumbromide). To quantify the state of cycloplegia, the abnormal pecking responses of cyclopleged chickens were studied. RESULTS: The authors found that cycloplegia could be maintained for 3 hours daily by corneal application of vecuroniumbromide. To ensure that visual exposure was restricted to the time period of cycloplegia, chickens were transferred to a 3-hour light/21-hour dark cycle. Control experiments showed that emmetropization was still functional under the changed light cycle. Strikingly, even with suppressed local-retinal growth control mechanisms (as indicated by the lack of deprivation myopia in a 6-OHDA injected group of chickens with occluders) and paralysis of accommodation, the eyes compensated for the defocus imposed by spectacles by changing their axial growth rates to be similar to those of eyes with functional accommodation. CONCLUSIONS: The findings show that the ciliary muscle and the activity of the iris sphincter muscle are not involved in emmetropization in chicks. If accommodation mediates the growth effects with lenses, it must happen via another pathway. Based on previous results, the authors propose that either the choroidal nerves from the ciliary ganglion to the choroid are important or that another yet unknown pathway from the Edinger Westphal nucleus to the eye transmits the necessary information.

Flicker parameters are different for suppression of myopia and hyperopia.

Axial eye growth rates in the chicken are controlled by local retinal image-processing circuits. These circuits quantify the loss of contrast for different spatial frequencies and promote axial eye growth rates in correlation with the amount of retinal image degradation ("deprivation myopia"). They also distinguish whether the plane of focus lies in front of or behind the retina. How the sign of defocus is detected still remains unclear. Cues from chromatic aberration are not important. In an attempt to isolate retinal circuits controlling the development of myopia or hyperopia, young chickens were raised in flickering light of different frequencies (12 and 6 Hz) and duty cycles (4-75%) produced by rotating chopper disks. The effects of flickering light on refractive errors and change in axial growth rates induced by translucent occluders or defocusing lenses were measured by infrared retinoscopy and A-scan ultrasound, respectively. Retinal electrical activity was evaluated by flicker ERG after matching flicker parameters and stimulation brightness at retinal surface. Changes in retinal and vitreal dopamine content caused by flicker in occluded and normal eyes were determined by HPLC-ECD. Strikingly, suppression of myopia occurred for similar flicker parameters, whether induced by translucent occluders ("deprivation") or negative lenses ("defocus"). The degree to which myopia was suppressed was correlated with the duration of flicker dark phase and with the ERG amplitude. In contrast, suppression of hyperopia did not correlate with these parameters. We conclude that two different retinal circuits with different temporal characteristics are involved in the processing of hyperopic defocus/deprivation and of myopic defocus, the first one dependent on flicker ERG amplitude. However, we did not find any correlation between the rate of dopamine release and the degree of inhibition of deprivation myopia in flickering light.

[Physiological functional evaluation of retinal implants in animal models]. / Physiologische Funktionsprüfungen von Retinaimplantaten an Tiermodellen.

Retinal implants can--by electrical stimulation--create visual impressions in people with certain kinds of degenerative retinal diseases (e.g. Retinitis Pigmentosa). Electrically evoked potentials in the retina must be transferred into the visual cortex in an orderly manner, a prerequisite for any kind of form- and movement-perception. In the current developmental stage the difficult investigations are performed in various animal models: isolated retinae of intact chicken and of RCS-rats (a model for Retinitis Pigmentosa), as well as in anesthetised rabbits, pigs and cats with intact retinae. Our investigations show that spatially selective ganglion-cell responses can be recorded following focal electrical stimulation, in healthy and as well in degenerated retinae. Registration of activities in area 17 of the visual cortex demonstrate that electrical retinal stimulation can indeed activate it.

[Subretinal microphotodiode array as replacement for degenerated photoreceptors?]. / Subretinales Mikrophotodioden-Array als Ersatz für degenerierte Photorezeptoren?

A survey is given on the status of developments, concerning a subretinal electronic microphotodiode array that aims at replacing degenerated photoreceptors. Various prototypes have been developed, tested, and implanted in various experimental animals up to 18 months. The fact that electrical responses were recorded from the visual cortex of pigs after electrical stimulation by subretinal electrodes and the fact that responses are also recorded in-vitro in degenerated rat retinae, shows the feasibility of this approach. However, there are a number of open questions concerning the biocompatibility, the long-time stability, and the type of transmitted image to be solved before application in patients can be considered.

Implantation of stimulation electrodes in the subretinal space to demonstrate cortical responses in Yucatan minipig in the course of visual prosthesis development.

PURPOSE: During the course of the development of visual prostheses, subretinal stimulation films were implanted in micropigs in order to prove the feasibility of subretinal electrical stimulation with subsequent cortical response. One aim was to demonstrate that epidural recording of visual evoked potentials is possible in the micropig. METHODS: Film-bound stimulation electrode arrays were placed in the subretinal space of micropigs. This enabled the retina to be stimulated subretinally. Since conventional visual evoked potential (VEP) measuring is virtually impossible in the pig from the neurosurgical point of view, epidural recording electrode arrays were positioned over the visual cortex as permanent electrodes. RESULTS: The feasibility of temporary implantation of film-bound stimulation electrode arrays was successfully demonstrated in the micropig model. On stimulation with monopolar voltage pulses (1000 to 3000 mV), reproducible epidural VEP measurements (5 to 10 micronV) were detected. CONCLUSIONS: The feasibility of subretinal stimulation of the retina was demonstrated in a retinal model that is similar to the human retina. This animal model therefore offers a suitable means of studying the tolerability of stimulation situations in the course of visual prosthesis development.

Effects of atropine on refractive development, dopamine release, and slow retinal potentials in the chick.

Atropine has previously been found to suppress visually induced myopia both in animals and humans. The mechanism of its action is unclear. We have studied its retinal effects in an in vitro preparation, using the retina-pigment epithelium-choroid complex of the chick eye. In vivo, deprivation myopia was induced by translucent goggles. Atropine solution was injected into the vitreous at two-day intervals. Dopamine release from the retina following atropine injection in vivo and from the in vitro retina preparation was quantified by HPLC-EC. In vitro preparations of the isolated chick retina-pigment epithelium-choroid were superfused with atropine. Light-induced potentials (local ERG), slow standing potentials from the retinal pigment epithelium/neural retina, and extracellular potassium concentrations were recorded. In line with previous findings, intravitreal injections of atropine (25 microg, 250 microg) reduced deprivation myopia in a dose-dependent manner. Atropine increased the release of the neurotransmitter dopamine into the superfusate in vitro at 100-500 microM and into the vitreous in vivo at 250 microg. Before an increase was measured in the vitreous, the retinal dopamine content was elevated. In concentrations equivalent to the intravitreal concentration to suppress myopia in vivo (200-800 microM), atropine induced spreading depression (SD) in the in vitro preparation. In contrast, muscarinic agonists, acetylcholine and pilocarpine, did not induce SD. Atropine reduced the ERG b- and d-wave, led to damped oscillations of RPE potentials, and reversed the ERG c-wave. Atropine suppressed myopia only at doses at which severe nonspecific side effects were observed in the retina. Atropine seems to intrude massively into the vital functions of the retina as indicated by the occurrence of SD. We conclude that atropine, by inducing SD, boosts neurotransmitter release from cellular stores, which may cancel out a presumed retinal signal that controls eye growth and through this, myopia.

Light-induced changes of extracellular ions and volume in the isolated chick retina-pigment epithelium preparation.

To better understand the mechanisms of extracellular space volume regulation and their possible effects on retinal function, light-induced changes in the concentrations of the principal extracellular ions (Na+, K+, Ca2+, and Cl-) were measured with ion-sensitive microelectrodes in the chick retina-pigment epithelium-choroid preparation. Changes of extracellular space volume were assessed by measuring the concentration of an impermeant marker, tetramethylammonium. In the inner retina, transient ON/OFF Na+ decrease was about twice as large as K+ increase, and the charge difference was compensated by a decrease in Cl- concentration. The ion changes were accompanied by extracellular space-volume decreases here. In the subretinal space, [Na+]o increase was about twice as large as K+ decrease, yet [Cl-]o, also decreased; this was accompanied by a sustained extracellular space-volume increase. The ionic changes in the inner retina are consistent with a model of extracellular space-volume regulation which assumes that neuronal depolarization causes net uptake of NaCl, cell swelling, and extracellular space shrinkage. However, to prevent the apparent violation of electroneutrality in the subretinal space, our simple model should be expanded to include the involvement of unidentified anion(s). Substantial changes in the subretinal space volume may influence interaction between the neural retina and pigment epithelium. Among ionic changes, only the light-induced [K+]o decrease around the photoreceptors and the [Ca2+]o increase near the photoreceptor bodies and synaptic terminals are large enough (-25% and 7.5%, respectively) to be likely candidates for integrated intercellular signaling.

Studies on the feasibility of a subretinal visual prosthesis: data from Yucatan micropig and rabbit.

BACKGROUND: To estimate the feasibility of the subretinal concept of a visual prosthesis, animal models and prototypes, each representing a certain aspect of the final prosthesis, were utilised to test for requirements for such a medical device: (1) the ability to elicit--by electrical stimulation--event-related central activity in the central visual system, and (2) the long-term biocompatibility and biostability of the implant within the subretinal space. METHODS: (1) In rabbit and Yucatan minipig, cortical evoked potentials were recorded with chronically implanted epidural electrodes during stimulation with light flashes as well as during electrical stimulation in the subretinal space. Voltage pulses ranging from -3 V to +3 V were applied via an acutely implanted electrode array on a wired prototype. (2) For biocompatibility studies a silicon-based micro-photodiode array (MPDA) was used that closely resembled the design and composition of the final prosthesis. Fourteen months after implantation, angiography was performed and the histological findings of the retina in the immediate vicinity of the implant were evaluated. RESULTS: (1) In both rabbit and minipig, subretinal electrical stimulation resulted in evoked cortical potentials that were comparable to visual evoked potentials. The lowest threshold levels for the subretinal stimulation were 0.6 V for rabbits and 2 V for minipigs. (2) Long-term stability of an implanted MPDA and its biocompatibility were proven for a postoperative period of 14 months. CONCLUSIONS: Data from animal experiments with certain prototypes of the final prosthesis suggest the feasibility of the concept of a subretinal visual prosthesis: Both requirements were met: (1) the functioning of the subretinal stimulation and (2) the biocompatibility of the MPDA implant.