Isolated Rabbit Retina Research Articles

Electric stimulus duration alters network-mediated responses depending on retinal ganglion cell type

Objective. To improve the quality of artificial vision that arises from retinal prostheses, it is important to bring electrically-elicited neural activity more in line with the physiological signaling patterns that arise normally in the healthy retina. Our previous study reported that indirect activation produces a closer match to physiological responses in ON retinal ganglion cells (RGCs) than in OFF cells (Im and Fried 2015 J. Physiol. 593 3677–96). This suggests that a preferential activation of ON RGCs would shape the overall retinal response closer to natural signaling. Recently, we found that changes to the rate at which stimulation was delivered could bias responses towards a stronger ON component (Im and Fried 2016a J. Neural Eng. 13 025002), raising the possibility that changes to other stimulus parameters can similarly bias towards stronger ON responses. Here, we explore the effects of changing stimulus duration on the responses in ON and OFF types of brisk transient (BT) and brisk sustained (BS) RGCs. Approach. We used cell-attached patch clamp to record RGC spiking in the isolated rabbit retina. Targeted RGCs were first classified as ON or OFF type by their light responses, and further sub-classified as BT or BS types by their responses to both light and electric stimuli. Spiking in targeted RGCs was recorded in response to electric pulses with durations varying from 5 to100 ms. Stimulus amplitude was adjusted at each duration to hold total charge constant for all experiments. Main results. We found that varying stimulus durations modulated responses differentially for ON versus OFF cells: in ON cells, spike counts decreased significantly with increasing stimulus duration while in OFF cells the changes were more modest. The maximum ratio of ON versus OFF responses occurred at a duration of ~10 ms. The difference in response strength for BT versus BS cells was much larger in ON cells than in OFF cells. Significance. The stimulation rates preferred by subjects during clinical trials are similar to the rates that maximize the ON/OFF response ratio in in vitro testing (Im and Fried 2016a J. Neural Eng. 13 025002). Here, we determine the stimulus duration that produces the strongest bias towards ON responses and speculate that it will further enhance clinical effectiveness.

Responses of Rabbit Retinal Ganglion Cells to Subretinal Electrical Stimulation Using a Silicon-Based Microphotodiode Array

With subretinal prostheses, retinal ganglion cells (RGCs) are activated by electrical stimulation of the retinal neural network. The aim of this study was to evaluate the efficacy of silicon-based solar cells in evoking RGC responses by electrically stimulating the photoreceptor side of an isolated retina. A light-bleached retina of an adult New Zealand White rabbit was placed with its photoreceptor side down onto a silicon chip that consisted of a 4 × 4 microphotodiode array (MPDA). The stimulating current was elicited by activating the solar cell with a 532-nm laser light source. Responses of the ON and OFF alpha RGCs on electrical stimulation were recorded extracellularly. Recorded RGCs were then injected with 4% N-(2-aminoethyl)-biotinamide hydrochloride to allow cell type identification. Using a design that includes a circumvented ground electrode, the authors successfully evoked spiking responses by the ON and OFF alpha RGCs in an isolated rabbit retina using low light power to activate the MPDA (equivalent to 39 μC/cm(2)). The charge density-dependent response and the frequency-dependent pair-pulse suppression were characterized. The spike latency of the RGC responses triggered by electrical stimulation was equivalent to the latency of its light response, which supports the hypothesis that the activation is mediated by the retinal neural network. Reliable activation of RGCs by electrical stimulation in vitro using an MPDA demonstrates the feasibility of developing solar cell-based subretinal prostheses that potentially could be developed into a power-free device able to restore vision.

Masked excitatory crosstalk between the ON and OFF visual pathways in the mammalian retina

A fundamental organizing feature of the visual system is the segregation of ON and OFF responses into parallel streams to signal light increment and decrement. However, we found that blockade of GABAergic inhibition unmasks robust ON responses in OFF α-ganglion cells (α-GCs). These ON responses had the same centre-mediated structure as the classic OFF responses of OFF α-GCs, but were abolished following disruption of the ON pathway with L-AP4. Experiments showed that both GABA(A) and GABA(C) receptors are involved in the masking inhibition of this ON response, located at presynaptic inhibitory synapses on bipolar cell axon terminals and possibly amacrine cell dendrites. Since the dendrites of OFF α-GCs are not positioned to receive excitatory inputs from ON bipolar cell axon terminals in sublamina-b of the inner plexiform layer (IPL), we investigated the possibility that gap junction-mediated electrical synapses made with neighbouring amacrine cells form the avenue for reception of ON signals. We found that the application of gap junction blockers eliminated the unmasked ON responses in OFF α-GCs, while the classic OFF responses remained. Furthermore, we found that amacrine cells coupled to OFF α-GCs display processes in both sublaminae of the IPL, thus forming a plausible substrate for the reception and delivery of ON signals to OFF α-GCs. Finally, using a multielectrode array, we found that masked ON and OFF signals are displayed by over one-third of ganglion cells in the rabbit and mouse retinas, suggesting that masked crossover excitation is a widespread phenomenon in the inner mammalian retina.

Dendritic morphology and tracer-coupling pattern of physiologically identified transient uniformity detector ganglion cells in rabbit retina

Transient uniformity detectors (UDs) are a unique type of retinal ganglion cell (RGC) whose maintained firing is transiently suppressed by all types of visual stimuli. In this study, we have characterized the dendritic morphology and tracer-coupling pattern of UDs that were labeled by loose-seal electroporation of Neurobiotin following functional identification in the isolated rabbit retina. The UDs have a bistratified dendritic tree, branching near the margins of the inner plexiform layer in stratum 1 (part of the OFF sublamina) and stratum 4/5 (part of the ON sublamina). Characteristically, many of the distal dendrites in the OFF arbor do not terminate there but dive recurrently back to the ON arbor. As a consequence, the ON dendritic arbor is usually twice as large as the OFF dendritic arbor in area. The UDs sometimes show homologous tracer coupling to neighboring RGCs with the same morphology, and from this material, we estimate that the UDs have a threefold dendritic field overlap and a maximum density of ~100 cells/mm2 on the peak visual streak, accounting for ~2% of RGCs in rabbit retina. The UDs also show strong heterologous tracer coupling to a novel type of amacrine cell that costratifies with the ON arbor of the UD. Consistent with their unistratified medium-field morphology, these St4/5 amacrine cells appear to be GABAergic: their somata are immunopositive for GABA but immunonegative for glycine and glycine transporter 1. We compare the dendritic morphology of the UDs to that of other types of bistratified RGCs described in rabbit retina and note that the stratification levels and distinctive recurrent dendrites closely resemble those of the "ON bistratified diving" RGCs. This raises the possibility that there are two types of RGCs with distinctive physiological properties that have almost identical bistratified dendritic morphologies.

Responses of ganglion cells to repetitive electrical stimulation of the retina

Retinal ganglion cells (RGCs) can be activated electrically either directly or indirectly (via the retinal neural network). Previous studies have shown that RGCs can follow high stimulus rates (⩾200 pulses s−1) when directly activated. In the present study, we investigated how well RGCs can follow repetitive stimulation of the neural network. We studied the responses (spike activity) of RGCs in isolated rabbit retina to stimulation with paired pulses applied at different interpulse intervals and trains of pulses applied at different frequencies. We found that the response amplitude of a RGC to a current pulse applied soon (⩽400 ms) after a preceding current pulse is diminished. This depression in response amplitude became greater as the interval between pulses became shorter. At an interpulse interval of 15 ms (shortest tested), the response amplitude to the second current pulse was reduced on average 94%. When a train of ten stimulus pulses was applied, further depression was observed, particularly at high stimulation frequencies. The depression with each successive pulse was relatively moderate compared to the depression to the second pulse. The results of this study have implications for the design of electrical stimulation strategies in a retinal prosthesis.

Electrical Stimulation in Isolated Rabbit Retina

Experiments were conducted to assess the effect of stimulating electrode parameters (size, position, and waveform shape) on electrically elicited ganglion cell action potentials from isolated rabbit retina. Thirty-eight isolated rabbit retinas were stimulated with bipolar stimulating electrodes (either 125 or 25 microm in diameter) positioned on either the ganglion or the photoreceptor side. Recording electrodes were placed between the optic disc and the stimulating electrodes. Cathodic-first, biphasic, current waveforms of varying pulse durations (0.1, 0.5, 1 ms) were used. For the four conditions tested (125-electrode and 25-microm electrode, ganglion cell, and photoreceptor positions) threshold currents ranged from 6.7 to 23.6 microA, depending on location and pulse duration. With 1-ms pulse duration, no statistically significant difference was seen between threshold currents when either size electrode was used to stimulate either the ganglion cell side or the photoreceptor side. For all groups, the threshold currents using the 1-ms pulse were lower than those using 0.1 ms, but the 0.1-ms pulses used less charge. These experiments provide a number of valuable insights into the relative effects of several stimulation parameters critical to the development of an implanted electronic retinal prosthesis.

Optophysiology: Depth-resolved probing of retinal physiology with functional ultrahigh-resolution optical coherence tomography

Noncontact, depth-resolved, optical probing of retinal response to visual stimulation with a <10-microm spatial resolution, achieved by using functional ultrahigh-resolution optical coherence tomography (fUHROCT), is demonstrated in isolated rabbit retinas. The method takes advantage of the fact that physiological changes in dark-adapted retinas caused by light stimulation can result in local variation of the tissue reflectivity. fUHROCT scans were acquired from isolated retinas synchronously with electrical recordings before, during, and after light stimulation. Pronounced stimulus-related changes in the retinal reflectivity profile were observed in the inner/outer segments of the photoreceptor layer and the plexiform layers. Control experiments (e.g., dark adaptation vs. light stimulation), pharmacological inhibition of photoreceptor function, and synaptic transmission to the inner retina confirmed that the origin of the observed optical changes is the altered physiological state of the retina evoked by the light stimulus. We have demonstrated that fUHROCT allows for simultaneous, noninvasive probing of both retinal morphology and function, which could significantly improve the early diagnosis of various ophthalmic pathologies and could lead to better understanding of pathogenesis.

Gap junctional coupling and connexin immunoreactivity in rabbit retinal glia

Gap junctions provide a pathway for the direct intercellular exchange of ions and small signaling molecules. Gap junctional coupling between retinal astrocytes and between astrocytes and Müller cells, the principal glia of vertebrate retinas, has been previously demonstrated by the intercellular transfer of gap-junction permeant tracers. However, functional gap junctions have yet to be demonstrated between mammalian Müller cells. In the present study, when the gap-junction permeant tracers Neurobiotin and Lucifer yellow were injected into a Müller cell via a patch pipette, the tracers transferred to at least one additional cell in more than half of the cases examined. Simultaneous whole-cell recordings from pairs of Müller cells in the isolated rabbit retina revealed electrical coupling between closely neighboring cells, confirming the presence of functional gap junctions between rabbit Müller cells. The limited degree of this coupling suggests that Müller cell-Müller cell gap junctions may coordinate the functions of small ensembles of these glial cells. Immunohistochemistry and immunoblotting were used to identify the connexins in rabbit retinal glia. Connexin30 (Cx30) and connexin43 (Cx43) immunoreactivities were associated with astrocytes in the medullary ray region of the retinas of both pigmented and albino rabbits. Connexin43 was also found in Müller cells, but antibody recognition differed between astrocytic and Müller cell connexin43.

Electrical stimulation of isolated retina with microwire glass electrodes

The development of high-resolution retinal prostheses fabricated from silicon wafers presents an interesting problem: how to electrically bridge the space between the flat silicon wafer and the curved retinal surface. One potential “bridge” is a microwire glass electrode. In this paper we present our results in evaluating microwire glass electrodes. We stimulated isolated rabbit retina ( n=5) with a 0.0256 cm 2 microwire electrode. The current and pulse duration were varied from 498 to 1660 μA and 0.1 to 3 ms, respectively. We found that short pulses produced more spikes per coulomb and longer pulses produced more spikes per milliamp. The optimal pulse duration range of 0.7–1 ms was identified as a compromise between the advantages of short and long pulses. Stimulation of isolated rabbit retina with microwire glass results in consistent neuronal spike formation at safe charge density, 20.7±4.3 μC/cm 2. We also examined the response of retinas ( n=6) to stimulation with a smaller microwire electrode, 0.0002 cm 2. We found that less current was required (15 μA versus 756 μA) for a 1 ms pulse, but at the expense of greater charge density (75 μC/cm 2 versus 29.5 μC/cm 2). Nonetheless, a 128-fold reduction in area resulted in only a 2.7-fold increase in charge density required for a 1 ms pulse duration. The results presented here indicate that microwire glass can be used as a neural stimulating electrode to bridge the gap between flat microelectronic stimulator chips and curved neuronal tissue.

Electrical stimulation of isolated retina with microwire glass electrodes

The development of high-resolution retinal prostheses fabricated from silicon wafers presents an interesting problem: how to electrically bridge the space between the flat silicon wafer and the curved retinal surface. One potential “bridge” is a microwire glass electrode. In this paper we present our results in evaluating microwire glass electrodes. We stimulated isolated rabbit retina (n=5) with a 0.0256 cm2 microwire electrode. The current and pulse duration were varied from 498 to 1660 μA and 0.1 to 3 ms, respectively. We found that short pulses produced more spikes per coulomb and longer pulses produced more spikes per milliamp. The optimal pulse duration range of 0.7–1 ms was identified as a compromise between the advantages of short and long pulses. Stimulation of isolated rabbit retina with microwire glass results in consistent neuronal spike formation at safe charge density, 20.7±4.3 μC/cm2. We also examined the response of retinas (n=6) to stimulation with a smaller microwire electrode, 0.0002 cm2. We found that less current was required (15 μA versus 756 μA) for a 1 ms pulse, but at the expense of greater charge density (75 μC/cm2 versus 29.5 μC/cm2). Nonetheless, a 128-fold reduction in area resulted in only a 2.7-fold increase in charge density required for a 1 ms pulse duration. The results presented here indicate that microwire glass can be used as a neural stimulating electrode to bridge the gap between flat microelectronic stimulator chips and curved neuronal tissue.

The effect of GABA and the GABA-uptake-blocker NO-711 on the b-wave of the ERG and the responses of horizontal cells to light.

The effects of GABA in the retina have now become of special interest because the anti-epileptic drug vigabatrin, a GABA analogue, can cause visual field loss in humans. Vigabatrin inhibits the GABA-aminotransferase, which finally results in GABA accumulation in the extracellular space. The b-wave of the electroretinogram (ERG), which originates partly in on-bipolar cells, is influenced by both GABAergic horizontal cells (HCs) and GABAergic amacrine cells (ACs). Their influences, however, are difficult to separate. In an attempt to isolate the effect of GABAergic ACs, use has been made of the specific effect of the GABA-uptake-blocker NO-711, which blocks only the GABA transporter GAT1 of GABAergic ACs. The ERG and the intracellular responses of HCs to light were recorded in the isolated rabbit retina, and the effects of GABA and NO-711, when added separately to the superfusate, were determined. GABA reduced significantly both the light responses of HCs and the b-wave. NO-711 enlarged the b-wave drastically, but did not affect the responses of HCs to light. An increase in the extracellular GABA concentration decreases the b-wave; an impairment of the function of ACs increases the b-wave. These conditions are discussed in the context of the lack of consistent changes to the b-wave during therapy with vigabatrin.

Mechanism of ganciclovir uptake by rabbit retina and human retinal pigmented epithelium cell line ARPE-19

Purpose. The objective of this study was to elucidate the mechanism of ganciclovir uptake by the rabbit retina and the human retinal pigmented epithelium cell line ARPE-19.Materials and methods. [3H]Adenine, [3H]adenosine, [3H]thymidine, and [3H]ganciclovir were used to elucidate the mechanism of ganciclovir uptake by the ARPE-19 cell line and the isolated rabbit retinal tissue. Uptake studies using ARPE-19 cell line and isolated rabbit retina were carried out at 37°C and 25°C, respectively, for 5 min.Results. Uptake of [3H]adenine by ARPE-19 cells decreased by 95% in the presence of unlabeled adenine. Other nucleobases such as guanine, thymine, and uracil and the nucleosides adenosine, guanosine, thymidine, uridine, and inosine also reduced uptake of [3H]adenine by the ARPE-19 cells. Although [3H]adenosine and [3H]thymidine uptake was inhibited by nucleosides, nucleobases did not demonstrate any inhibitory effect, indicating that nucleosides can only bind to the nucleobase transporter but are not translocated by it. Uptake of the nucleosides and nucleobases by the ARPE-19 cells was sodium and pH independent. [3H]adenosine and [3H]thymidine uptake by the ARPE-19 cells was inhibited by nanomolar quantities of nitrobenzylthioinosine. Uptake of [3H]adenine by the isolated rabbit retina was drastically reduced in the presence of unlabeled adenine. Unlabeled thymidine and guanosine, and removal of sodium from the uptake medium, inhibited uptake of [3H]thymidine by the rabbit retina. Nucleosides, nucleobases, and unlabeled ganciclovir did not exhibit any inhibitory effect on [3H]ganciclovir uptake by the isolated rabbit retina or ARPE-19 cells.Conclusions. Our results indicate that although the rabbit retina and the ARPE-19 cell line express nucleoside and nucleobase transporters, translocation of ganciclovir does not involve any carrier-mediated transport process. Rather, ganciclovir uptake by the rabbit retina and ARPE-19 cells is governed primarily by passive diffusion.

The effects of GABA and vigabatrin on horizontal cell responses to light and the effect of vigabatrin on the electroretinogram.

When used as an antiepileptic drug in humans vigabatrin, which is a GABA analogue and an inhibitor of GABA-aminotransferase, often causes peripheral visual field loss. This could result from increases in endogenous GABA levels. Accordingly we have investigated the effects of GABA on horizontal cells (HCs) of the rabbit retina, and of vigabatrin, when applied for only a few minutes, on HCs and on the electroretinogram (ERG). The intracellular HC and ERG records were first obtained from isolated rabbit retinas during perfusion with a physiological solution. The perfusate was then changed to one containing GABA (2 mM) or vigabatrin (25, 40 or 150 microM) for at least 5 min, and then returned to the control solution. 2 mM GABA significantly but reversibly reduced the light responses of HCs elicited by diffuse light (at -4 log intensity) to 52 +/- 17% (SD, n = 7). Vigabatrin had no significant effect on the light responses of HCs (n = 7), and no effect on the b-wave (n = 4), but the PIII-component of the ERG was slightly but significantly reduced to 84 +/- 5% (SD, n = 5). The high dosage of GABA needed to affect the light responses of HCs could be due to strong GABA uptake systems in the intact rabbit retina. It is, however, possible that in humans receiving long-term treatment with vigabatrin, high levels of GABA occur because of the inhibition of GABA- aminotransferase. It seems, from these observations, that neurons like on-bipolar cells, which are contributors to the b-wave, and HCs are uninfluenced by vigabatrin in short-term experiments. The slightly reduced slow PIII-component, however, indicates an influence on the glial Müller cells which are the main contributors to the slow PIII-component.

The influence of HEPES on light responses of rabbit horizontal cells

HEPES-buffered solutions, mostly used in studies of isolated cells, and bicarbonate-buffered solutions, mostly used in studies of isolated retinal tissues, have both been used to superfuse an isolated rabbit retina preparation. The responses of horizontal cells (HCs) to light, detected by intracellular microelectrodes filled with Lucifer Yellow, were recorded. Buffering of the superfusate with 100% HEPES completely, but reversibly, abolished the responses of A-type HCs, and is not, therefore, suitable for studies on isolated rabbit retinas. The responses remained when buffering was partially with HEPES and partially with bicarbonate, but were changed: in A-type HCs the overshoot was reduced and the afterpotential was increased. The overshoot may be caused by feedback of HCs on the cones and might be dependent on pHi at the synaptic structure between HCs and photoreceptors.

Low b-wave amplitudes in a strain of rabbits with a pigment epithelium defect

When preparing isolated rabbit retinas we found in some animals fundi which were not uniformly dark but had abnormal areas of red coloration. The in situ electroretinograms (ERG) of 82 rabbits recorded after 1 h of dark adaptation were checked for abnormalities indicative of a degenerative disorder. The ERGs of eight rabbits with small dark adapted b-waves (≤250 μV) were re-recorded and their b-waves found to decline with time. The greatest reduction, in three rabbits, was ≥150 μV over 2.5 years. After 1 year, however, the light adapted b-waves were similar to those of rabbits with normal dark adapted b-waves. The majority of the progeny of these rabbits also had small b-waves, which became still smaller in 2 years. Ultrastructural studies of two rabbit retinas of the first generation showed pathological changes of the pigment epithelium (Wrigstad, Hanitzsch & Nilsson, Ultrastructural and electrophysiological studies of the retina and the retinal pigment epithelium in rabbits with low b-wave amplitudes, in preparation). Evidently there is an inheritable defect in the pigment epithelium which first impairs the rod pathway.

Mapping glutamatergic drive in the vertebrate retina with a channel-permeant organic cation.

Patterns of neuronal excitation in complex populations can be mapped anatomically by activating ionotropic glutamate receptors in the presence of 1-amino-4-guanidobutane (AGB), a channel-permeant guanidinium analogue. Intracellular AGB signals were trapped with conventional glutaraldehyde fixation and were detected by probing registered serial thin sections with anti-AGB and anti-amino acid immunoglobulins, revealing both the accumulated AGB and the characteristic neurochemical signatures of individual cells. In isolated rabbit retina, both glutamate and the ionotropic glutamate receptor agonists alpha-amino-3-hydroxyl-5-methylisoxazole-4-propionic acid (AMPA), kainic acid (KA), and N-methyl-D-aspartic acid (NMDA) activated permeation of AGB into retinal neurons in dose-dependent and pharmacologically specific modes. Horizontal cells and bipolar cells were dominated by AMPA/KA receptor activation with little or no evidence of NMDA receptor involvement. Strong NMDA activation of AGB permeation was restricted to subsets of the amacrine and ganglion cell populations. Threshold agonist doses for the most responsive cell groups (AMPA, 300 nm; KA, 2 microM; NMDA, 63 microm; glutamate, 1 mM) were similar to values obtained from electrophysiological and neurotransmitter release measures. The threshold for activation of AGB permeation by exogenous glutamate was shifted to <200 microM in the presence of the glutamate transporter antagonist dihydrokainate, indicating substantial spatial buffering of extracellular glutamate levels in vitro. Agonist-activated permeation of AGB into neurons persisted under blockades of Na+ -dependent transporters, voltage-activated Ca2+ and Na+ channels, and ionotropic gamma-aminobutyric acid and glycine receptors. Cholinergic agonists evoked no permeation.

Magnesium plus mexiletine inhibit energy usage and protect retinas against ischemia.

We determined whether exogenous Mg2+ and/or mexiletine (Mex), which are reported to be neuroprotective agents, reduced neuronal energy requirements and protected against ischemia, using isolated rabbit retinas. Under non-ischemic conditions, Mex (300 microM) and the combination of Mg2+ (1 mM) plus Mex (300 microM) significantly reduced glucose utilization, by 19% and 31%, respectively. The combination of Mg2+ plus Mex, but not either agent alone, significantly reduced lactate production (by 18%; p < 0.05). When added during 2 h of ischemia (simulated by the reduction of oxygen from 95% to 15% and of glucose from 6 mM to 1 mM), Mg2+ plus Mex improved the recovery of glucose utilization (p < 0.01), lactate production (p < 0.05) and neuronal function (p < 0.05) for 3 h following return to control (post-ischemia/recovery) conditions. Thus reducing energy demands by blocking functions during temporary ischemia, protects neurons from irreversible functional damage.

ON direction-selective ganglion cells in the rabbit retina: dendritic morphology and pattern of fasciculation.

ON direction-selective (DS) ganglion cells were identified by electrophysiological recordings in DAPI labeled, isolated rabbit retinas. Their responses to a flashing spot were sustained. Their responses to moving stimuli were strong in the preferred direction and weak in the null direction. Injection of the recorded cells with Lucifer yellow revealed that the cells had a distinct dendritic morphology, consistent with that described previously (Buhl & Peichl, 1986; Amthor et al., 1989; Famiglietti, 1992a). When neighboring cells were injected, an extensive dendritic co-fasciculation was observed. The pattern of fasciculation restricts the possible synaptic connections of the ON DS cell.

Mosaic arrangement of ganglion cell receptive fields in rabbit retina.

The arrangement of ganglion cell receptive fields on the retinal surface should constrain several properties of vision, including spatial resolution. Anatomic and physiological studies on the mammalian retina have shown that the receptive fields of several types of ganglion cells tile the retinal surface, with the degree of receptive field overlap apparently being similar for the different classes. It has been difficult to test the generality of this arrangement, however, because it is hard to sample many receptive fields in the same preparation with conventional single-unit recording. In our experiments, the response properties and receptive fields of up to 80 neighboring ganglion cells in the isolated rabbit retina were characterized simultaneously by recording with a multielectrode array. The cells were divided into 11 classes on the basis of their characteristic light responses and the temporal structures of their impulse trains. The mosaic arrangement of receptive fields for cells of a given class was examined after the spatial profile of each receptive field was fitted with a generalized Gaussian surface. For eight cell classes the mosaic arrangement was similar: the profiles of neighboring cells approached each other at the 1-sigma border. Thus field centers were 2 sigma apart. The layout of fields for the remaining three classes was not well characterized because the fields were poorly fitted by a single Gaussian or because the cells responded selectively to movement. The 2-sigma center-center spacing may be a general principle of functional organization that minimizes spatial aliasing and confers a uniform spatial sensitivity on the ganglion cell population.

In vivo and in vitro experiments show that betaxolol is a retinal neuroprotective agent

The aim of the study was to determine whether betaxolol is a neuroprotective agent and can therefore slow down the changes seen in the retina following ischaemia/reperfusion. Ischaemia was induced in one rat eye by raising the intraocular pressure for 45 min. Three days later electroretinograms were recorded from both eyes and the retinas were examined immunohistochemically for the localisation of calretinin and choline acetyltransferase (ChAT) immunoreactivities. The effect of glutamate agonists, hypoxia or experimental ischaemia was examined on the GABA immunoreactivity, lactate dehydrogenase (LDH) and internal calcium levels ([Ca 2+] i) of the isolated rabbit retina, rat cortical cultures and chick retinal cell cultures respectively. Betaxolol was tested to see whether it can attenuate the influence of the glutamate agonists, hypoxia or experimental ischaemia. Ischaemia for 45 min causes a change in the nature of the normal calretinin immunoreactivity, an obliteration of the ChAT immunoreactivity and a drastic reduction in the b-wave of the electroretinogram after 3 days of reperfusion. When betaxolol was injected i.p. into the rats before ischaemia and on the days of reperfusion the changes to the calretinin and ChAT immunoreactivities were reduced and the reduction of the b-wave was prevented. Rabbit retinas incubated in vitro in physiological solution lacking oxygen/glucose or containing the glutamate agonists kainate or NMDA caused a change in the nature of the GABA immunoreactivity. Inclusion of betaxolol partially prevented the changes caused by NMDA and lack of oxygen/glucose. Rat cortical cultures exposed to glutamate or hypoxia/reoxygenation resulted in a release of LDH. The release of the enzyme was almost completely attenuated when betaxolol was included in the culture medium. Kainate increased the [Ca 2+] i in chick retinal cultures, as measured with Indo-1. In a medium with sodium, this kainate-induced elevation of [Ca 2+] i was significantly reduced by betaxolol. The combined data show that betaxolol is a neuroprotective agent and attenuates the effects on the retina induced by raising the intraocular pressure to simulate an ischaemic insult as may occur in glaucoma. © 1997 Elsevier Science B.V. All rights reserved.