Psychophysical research has documented the existence of three processes in light adaptation: a fast subtractive process, a divisive process that is fast at light onset and slower at light offset, and a very slow subtractive process (Hayhoe et al., 1987). In the neural model developed here, the fast subtractive process is identified with horizontal cell feedback onto cones and the divisive process with amacrine cell feedback onto bipolar cells. The very slow subtractive process is identified with the modulatory feedback circuit from amacrines via interplexiform cells to horizontal cells. A nonlinear dynamical model is developed incorporating these aspects of retinal circuitry along with both ON- and OFF-center M and P pathways. This model is shown to account for many aspects of foveal light adaptation, including negative afterimage formation, and to explain a number of the physiological differences between M and P ganglion cells, including their differing contrast-response functions.

The expression of GABA in the human fetal (12–25 weeks of gestation), postnatal (five-month-old), and adult (35-year-old) retinas was investigated by immunohistochemistry. GABA expression was seen as early as 12 weeks in the undifferentiated cells of the inner neuroblast zone; a few optic nerve fiber layer axons were clearly labeled, suggesting that some of the stained cell bodies were prospective ganglion cells, others could be displaced amacrine cells. From 16–17 to 24–25 weeks, intense labeling was found in the amacrine, displaced amacrine, and some ganglion cells. During this time period, horizontal cells (identified by calbindin immunohistochemistry), undergoing migration (periphery) and differentiation (center), expressed GABA prominently. In the postnatal retina, some horizontal cells were moderately labeled, but very weakly in a few cells, in the adult. The Müller cells developed immunoreactivity first weakly at 12 weeks and then moderately from 16–17 weeks onward. The staining was also evident in the postnatal and adult retinas, showing labeled processes of these glial cells. Virtually no axons in the adult optic nerve and nerve fiber layer were stained; the staining was restricted to a few, large ganglion cells and displaced amacrine cells. Some amacrines were also labeled. The possibility that GABA might play a role in horizontal cell differentiation and maturation is highlighted. Other evidences suggest that GABA might play a role in metabolism during retinal development.

The organization of the primate nerve fiber layer and optic nerve head with respect to the positioning of central and peripheral axons remains controversial. Data were obtained from 32 human fetal retinae aged between 15 and 21 weeks of gestation. Crystals of the carbocyanine dyes, DiI or DiA, and fluorescence microscopy were used to identify axonal populations from peripheral retinal ganglion cells. Peripheral ganglion cell axons were scattered throughout the vitreal-scleral depth of the nerve fiber layer. Such a scattered distribution was maintained as the fibers passed through the optic nerve head and along the optic nerve. There was a rough topographic representation within the optic nerve head according to retinal quadrant such that both peripheral and central fibers were mixed within a wedge extending from the periphery to the center of the nerve. There was no indication that the fibers were reorganized in any way as they passed through the optic disc and into the nerve. The present results suggest that any degree of order present within the fiber layer and optic nerve is not an active process but a passive consequence of combining the fascicles of the retinal nerve fiber layer. Optic axons are not instructed to establish a retinotopic order and the effect of guidance cues in reordering fibers, particularly evident prechiasmatically and postchiasmatically, does not appear to be present within the nerve fiber layer or optic nerve head in humans.

The retinal pigment epithelium (RPE) of the quokka wallaby, Setonix brachyurus, grows and changes throughout life. To investigate factors that determine changes in the quokka RPE, we have examined topography of this tissue in experimentally enlarged eyes. Unilateral eyelid suture was conducted at the time of normal eye opening, postnatal day (P) 110, and animals were examined at 1 or 1½ years of age. The numbers and densities of RPE cells and the extent of multinucleation were compared with those in normal animals. Eyelid suture resulted in a 9.8% and 17.4% increase in retinal area at 1 and 1½ years, respectively; a significant degree of myopia was associated with this enlargement. Cell density topography in experimental eyes was not the same as in controls. Cells from central retina were disproportionately larger in the experimental than control eyes. However, the RPE cell topography in sutured eyes was not the same as that of aged retinae of a similar size. Notably, in sutured eyes there was no development of the high or highest cell densities seen in equatorial and temporal central RPE in aged retinae, respectively. Furthermore, the degree of cell enlargement in peripheral regions was slight compared with that observed in similar-sized, aged retinae. There was no increase in RPE cell number; rather, average cell area increased accompanied by no change or a slight decrease in RPE thickness. Consequently, overall volume of cells did not change significantly. The large number of multinucleate cells normally seen in aged animals was not observed in experimentally enlarged eyes, implying that an increase in cell volume may be the trigger for multinucleation.

This study sought to characterize the tracer coupling of regenerated amacrine cells in the retina of the goldfish and assess the integration of regenerated neurons into existing retinal circuits. Regeneration of new neurons from injury-induced progenitors was stimulated by surgically excising a small rectangular piece of retina. Several months after regeneration was complete, intracellular injections of Neurobiotin, a gap junction-permeant tracer, were made into single regenerated amacrine cells or nonregenerated (extant) amacrine cells lying outside the regenerated patch. Two groups of amacrine cells were injected: those that in normal retina are tracer coupled and a single type (the radiate amacrine cell) that is not. The data show that regenerated amacrine cells are tracer coupled to each other and to their homologous counterparts outside the patch of regenerated retina. Regenerated radiate cells possess morphologically abnormal dendrites, but these processes can extend out of regenerated retina into surrounding normal retina. Similarly, the dendrites of extant radiate cells, severed by the original lesion, can regenerate into the patch of regenerated retina. These results indicate that in the goldfish retina the cell-specific junctional circuitry present in normal retina is re-created in the regenerated retina, and suggest that regenerated neurons are functionally integrated into the existing retina.

In the rabbit retina, the nuclear dye, 4,6, diarnidino-2-phenylindole (DAPI), selectively labels a third type of amacrine cell, in addition to the previously characterized type a and type b cholinergic amacrine cells. In this study, these “DAPI-3” amacrine cells have been characterized with respect to their somatic distribution, dendritic morphology, and neurotransmitter content by combining intracellular injection of biotinylated tracers with wholemount immunocytochemistry. There are about 100,000 DAPI-3 amacrine cells in total, accounting for 2% of all amacrine cells in the rabbit retina, and their cell density ranges from about 130 cells/mm2 in far-peripheral retina to 770 cells/mm2 in the visual streak. The thin varicose dendrites of the DAPI-3 amacrine cells form a convoluted dendritic tree that is symmetrically bistratified in S1/S2 and S4 of the inner plexiform layer. Tracer coupling shows that the DAPI-3 amacrine cells have a fivefold dendritic-field overlap in each sublamina, with the gaps in the arborization of each cell being occupied by dendrites from neighboring cells. The DAPI-3 amacrine cells consistently show the strongest glycine immunoreactivity in the rabbit retina and they also accumulate exogenous [3H]-glycine to a high level. By contrast, the All amacrine cells, which are the best characterized glycinergic cells in the retina, are amongst the most weakly labelled of the glycine-immunopositive amacrine cells. The DAPI-3 amacrine cells costratify narrowly with the cholinergic amacrine cells and the On-Off direction-selective ganglion cells, suggesting that they may play an important role in movement detection.

Previous experiments in chickens have shown that dopamine released from the retina may be one of the messengers controlling the growth of the underlying sclera. It is also possible, however, that the apparent relationship between dopamine and myopia is secondary and artifactual. We have done experiments to assess this hypothesis. Using High Pressure Liquid Chromatography with electrochemical detection (HPLC-ED), we have asked whether changes in dopamine metabolism are restricted to the local retinal regions in which myopia was locally induced. Furthermore, we have measured the concentrations of biogenic amines separately in different fundal layers (vitreous, retina, choroid, and sclera) to find out how changes induced by “deprivation” (= removal of high spatial frequencies from the retinal image by translucent eye occluders which produce “deprivation myopia”) are transmitted through these layers. Finally, we have repeated the deprivation experiments after intravitreal application of the irreversible dopamine re-uptake blocker reserpine to see how suppression of dopaminergic transmission affects these changes. We found that (1) Alterations in retinal dopamine metabolism were indeed restricted to the retinal areas in which myopia was induced. (2) The retina was the major source of dopamine release with a steep gradient both to the vitreal and choroidal side. Vitreal content was about one-tenth, choroidal content about one-third, and scleral content about one-twentieth of that of the retina. (3) There was a drop by about 40% in vitreal dopamine, DOPAC (3,4-dihydroxyphenylacetic acid) and HVA (homovanilic acid) concentrations following deprivation which occurred already at a time where little changes could yet be seen in their total retinal contents. (4) Choroidal and scleral dopamine levels were not affected by deprivation, indicating that other messengers must relay the information to the sclera. (5) A single intravitreal injection of reserpine lowered dopamine and HVA levels in retina and vitreous for at least 10 days in a dose-dependent fashion and diminished or suppressed further effects of deprivation on these compounds. DOPAC levels continued to change upon deprivation even after reserpine injection (Fig. 3). Our results suggest that the release rates of dopamine from retinal amacrine cells can be estimated from vitreal dopamine concentrations; furthermore, they are in line with the hypothesis that there is an inverse relationship between dopamine release and axial eye growth rates. Although our experiments do not ultimately prove that dopamine has a functional role in the visual control of eye growth, they are in line with this notion.

We characterized the light response, morphology, and receptive-field structure of a distinctive amacrine cell type (Dacey, 1989), termed here the Al amacrine, by applying intracellular recording and staining methods to the macaque monkey retina in vitro. A1 cells show two morphologically distinct components: a highly branched and spiny dendritic tree, and a more sparsely branched axon-like tree that arises from one or more hillock-like structures near the soma and extends for several millimeters beyond the dendritic tree. Intracellular injection of Neurobiotin reveals an extensive and complex pattern of tracer coupling to neighboring A1 amacrine cells, to two other amacrine cell types, and to a single ganglion cell type. The A1 amacrine is an ON-OFF cell, showing a large (10–20 mV) transient depolarization at both onset and offset of a photopic, luminance modulated stimulus. A burst of fast, large-amplitude (Σ60 mV) action potentials is associated with the depolarizations at both the ON and OFF phase of the response. No evidence was found for an inhibitory receptive-field surround. The spatial extent of the ON-OFF response was mapped by measuring the strength of the spike discharge and/or the amplitude of the depolarizing slow potential as a function of the position of a bar or spot of light within the receptive field. Receptive fields derived from the slow potential and associated spike discharge corresponded in size and shape. Thus, the amplitude of the slow potential above spike threshold was well encoded as spike frequency. The diameter of the receptive field determined from the spike discharge was Σ10% larger than the spiny dendritic field. The correspondence in size between the spiking receptive field and the spiny dendritic tree suggests that light driven signals are conducted to the soma from the dendritic tree but not from the axon-like arbor. The function of the axon-like component is unknown but we speculate that it serves a classical output function, transmitting spikes distally from initiation sites near the soma.

The compound eye of the tobacco hornworm moth Manduca sexta contains green-, blue-, and ultraviolet-sensitive photoreceptors. Electroretinogram spectral-sensitivity measurements were recorded from different regions of the retina in order to broadly map the distribution of the three receptor types. The relative contribution of the three receptors to spectral-sensitivity curves was estimated by fitting theoretical curves based on the absorption spectra of the three rhodopsins. This analysis indicated that the dorsal retina is green and ultraviolet dichromatic, with green-sensitive cells greatly predominating. The ventral retina is trichromatic with a substantial population of blue- and ultraviolet-sensitive receptors. We previously showed that flower visitation for nectar feeding is mediated mainly by blue-sensitive cells. Their localization in the ventral retina seems an appropriate adaptation of the receptor mosaic, since the moths hover above flowers as they feed.

The perception of dynamic random-dot stereograms (RDS) depends on the physiological fusion of horizontally disparate binocular visual input. Thus, the use of RDS offers the possibility to study selectively cortical processing of visual information in man. We investigated the influence of horizontal disparity on the scalp topography of RDS evoked brain activity in 33 healthy subjects. Stereoscopic checkerboard patterns were presented in the center or lateralized in the left or right visual field with horizontal disparities changing at temporal frequencies of six or eight depth reversals/s using different disparity values ranging from 3.5 to 28 min of arc. In 11 subjects evoked potential fields were recorded from 16 electrodes, and 21 subjects participated in 30-channel recordings with electrodes located over the parietal and occipital brain areas. Stimulation frequency-related brain activity was obtained with all disparity values; however, with large or small disparities the potential field strength decreased significantly while largest responses were obtained with intermediate disparities. Significant differences were observed in RDS evoked brain activity when central and lateralized stimulus locations were compared. With lateral stimuli (extending from the fovea to 17.1-deg eccentricity) maximal amplitudes were obtained at larger disparities than with central stimuli. In addition there were pronounced differences between brain activity evoked with stimuli presented in the left or right visual field; however, there were very similar evoked potential signals recorded from electrodes located over the left and right hemispheres. Our findings indicate that the processing of disparity information with lateralized stimuli is different from the processing in the center of the visual field. In addition, lateralized stimulation yields a significant disparity tuning mainly with stereoscopic targets occurring to the right from the fixation point (but not with stimuli to the left) suggesting a functional difference between the visual half-fields.

The multi-input technique of Sutter and Tran (1992) yields multiple focal ERGs. The purpose here was to compare the components of this multifocal ERG to the components of the standard, full-field ERG. To record multifocal ERGs, an array of 103 hexagons was displayed on a monitor. Full-field (Ganzfeld) ERGs were elicited by flashes presented upon steady background fields. The latencies of two prominent subcomponents of the full-field ERG were altered by varying the intensity of the incremental flash or the intensity of the background field. By showing that similar manipulations of the multi-input parameters produce similar changes in latency, we were able to relate the components of the multifocal ERG to the components of the full-field ERG. The biphasic responses of the multifocal ERG appear to be generated by the same cells generating the a-wave and positive peaks of the full-field cone ERG.

Dopamine is one of the major neurotransmitters in the retina. It is released from amacrine and interplexiform cells into both inner (IPL) and outer (OPL) plexiform layers. Several dopaminergic actions are known to occur through D1 receptors (D1R) but the precise location of these receptors has not been established. An antibody that recognizes the intracytoplasmic C-terminal of the rat D1R was used to detect D1R, immunohistochemically, in rats (Wistar and RCS), mouse, hamster, and macaque monkey retinas. The OPL was heavily stained in each species, consistent with the known actions of dopamine on horizontal cells. Three to five bands were observed in the IPL, depending on species. Three were in the a sublayer, the outermost of which was close to the amacrine cell layer, and may represent the massive dopamine input to the AII rod-amacrine cells. As observed in mice, where bipolar cells are D1-immunoreactive, the band located in sublayer 3 of the IPL may contain cone-bipolar cell terminals. A band of D1R-immunoreactivity in the b sublayer of the IPL contains ON-bipolar cell terminals and a second site of interaction between dopaminergic cells and the AII amacrine cells. This sublayer was absent from the RCS rat retina, suggesting a severe impairment of the rod-driven pathway following rod degeneration in these mutant rats. Cells in the ganglion cell layer exhibited relatively heavy staining, and may be ganglion cells or displaced amacrine cells. Some extrasynaptic localizations of D1R in the retina are suggested.

Whole-cell voltage clamp in the retinal slice and intracellular current clamp in the intact retina were used to study inhibitory interactions in the inner plexiform layer. Picrotoxin or strychnine reduced inhibitory, light-evoked currents in a majority of ganglion cells. However, in nearly a third of the ganglion cells, each of these antagonists enhanced the inhibitory synaptic current. All inhibitory current was blocked by the addition of the other antagonist. This indicates a cross-inhibition between GABAergic and glycinergic feedforward pathways. Blocking of GABAARs with SR95531 shortened the time course of both excitatory and inhibitory synaptic currents in ganglion cells. Application of picrotoxin, which blocked both GABAARs and GABACRs, produced the opposite effect. Recordings in the intact retina indicated that the light responses of ON bipolar cells, sustained ON, and transient ON-OFF third-order neurons were all made more transient by SR95531 and made more sustained by picrotoxin. The data suggest that a GABAC feedback pathway to bipolar cells makes light responses more phasic and that this feedback is inhibited through a GABAAR pathway. Consequently, the balance between GABAAR and GABACR inhibition regulates the time course of inputs to ganglion cells.

The rod-driven, AII amacrine cells in the mammalian retina maintain homologous gap junctions with one another as well as heterologous gap junctions with on-cone bipolar cells. We used background illumination to study whether changes in the adaptational state of the retina affected the permeabilities of these two sets of gap junctions. To access changes in permeability, we injected single AII amacrine cells with the biotinylated tracer, Neurobiotin, and measured the extent of tracer coupling to neighboring AII cells and neighboring cone bipolar cells. We also measured the center-receptive field size of All cells to assess concomitant changes in electrical coupling. Our results indicate that in well dark-adapted retinas, AII cells form relatively small networks averaging 20 amacrine cells and covering about 75 μm. The size of these networks matched closely to the size of AII cell on-center receptive fields. However, over most of their operating range, AII cells formed dramatically larger networks, averaging 326 amacrine cells, which corresponded to an increased receptive-field size. As the retina was light adapted beyond the operating range of the AII cells, they uncoupled to form networks comparable in size to those seen in well dark-adapted retinas. Our results, then, indicate that the adaptational state of the retina has a profound effect on the extent of electrical coupling between AII amacrine cells. Although we observed light-induced changes in the number of tracer-coupled cone bipolar cells, these appeared to be an epiphenomenon of changes in homologous coupling between AII amacrine cells. Therefore, in contrast to the robust changes in AII–AII coupling produced by background illumination, our data provided no evidence of a light-induced modulation of coupling between AII cells and on-cone bipolar cells.

We recorded visual evoked potentials in response to square-wave contrast-reversal checkerboards undergoing a transition in the mean contrast level. Checkerboards were modulated at 4.22 Hz (8.45-Hz reversal rate). After each set of 16 cycles of reversals, stimulus contrast abruptly switched between a “high” contrast level (0.06 to 1.0) to a “low” contrast level (0.03 to 0.5). Higher contrasts attenuated responses to lower contrasts by up to a factor of 2 during the period immediately following the contrast change. Contrast-response functions derived from the initial second following a conditioning contrast shifted by a factor of 2–4 along the contrast axis. For low-contrast stimuli, response phase was an advancing function of the contrast level in the immediately preceding second. For high-contrast stimuli, response phase was independent of the prior contrast history. Steady stimulation for periods as long as 1 min produced only minor effects on response amplitude, and no detectable effects on response phase. These observations delineate the dynamics of a contrast gain control in human vision.

Morphology of the central retina and scotopic visual sensitivity were compared in juvenile albino and normally pigmented rainbow trout living under natural and reduced daylight. Outdoor albinos avoided exposing their eyes to direct sunlight, whereas normals were indifferent to it. After 4 months outdoors (Σ10,000 lux in albinos, Σ100,000 lux in normals), albinos had severely truncated or missing rod outer segments (ROS) and some missing rod ellipsoids, but normal numbers of photoreceptor nuclei and fully intact cones. Albino estimated ROS volume was only 7.1% of normal in July, but increased to 20% by the following February, mainly via an increase in numbers of ROS. However, in albinos moved indoors October 7 and exposed to 10–30 lux ambient daylight, both the number and length of ROS increased significantly, with estimated ROS volume reaching 95% of normal by 34 days. Albinos generally had more phagosomes (Σ3 X normal) and more macrophages (Σ2 X normal) in their outer retina. An optomotor reflex was used to define the effect of ROS volume on the ability to respond visually during dark adaptation. In July, albinos and normals from outdoor raceways (3 months) or indoor raceways (35 days) showed equal sensitivity after first being placed in darkness, but after 1 h in darkness, outdoor albinos with 6% of normal ROS volume were 2.0 log units less sensitive than indoor or outdoor normals, whereas indoor albinos with 53% of normal ROS volume were only 0.7 log units less sensitive. This verifies that most rod cell bodies of albino trout can persist without functional ROS in indirect sunlight, and can regrow functional outer segments in dim daylight. This finding is distinct from the extensive retinal light damage observed in albino rats exposed to more moderate cyclic light, in which entire rod cells degenerate early on.

A class 1 aldehyde dehydrogenase (ALDH) catalyzes oxidation of retinaldehyde to retinoic acid in bovine retina. We used immunocytochemistry and in situ hybridization to localize this enzyme in adult and fetal bovine retinas. Specific ALDH immunoreactivity was present in the cytoplasm of wide-field amacrine cells restricted in distribution to the dorsal part of the adult retina. The somata diameters ranged from ∼8 μ to ∼15 μ, and the cells increased in density from ∼125 cells/mm2 near the horizontal meridian to ∼425 cells/mm2 in the superior far periphery. The ALDH-positive cells had somata on both sides of the inner plexiform layer (IPL) and processes in two IPL strata. The majority of ALDH-positive cells were unreactive with antibodies against known amacrine cell enzymes and neurotransmitters, including GABA and glycine. The ALDH-positive amacrine cells also did not react with anti-cellular retinoic acid-binding protein, which was present in a subset of GABA-positive amacrine cells. In flat-mounted retinas processed by in situ hybridization, the larger ALDH-positive amacrine cells tended to be more heavily labeled. In addition to amacrine cells, Müller cell processes in the inner retina were weakly immunoreactive for ALDH; however, these glial cells did not contain ALDH mRNA. The pattern of ALDH expression in fetal bovine retinas was documented by immunocytochemistry. No ALDH reactivity was found before 5.5 months; for the remainder of the fetal period, ALDH immunoreactivity was present in amacrine cells similar to those in adult retina. The ALDH-positive amacrine cells in bovine retina are novel, being limited in distribution to the dorsal retina and unlabeled with other amacrine cell-specific markers. Identification of ALDH in amacrine cells provides additional evidence that cells of the inner retina are involved in retinoid metabolism.