We have developed an optical method to monitor the activity of the ON and OFF channels in the anuran retina. The change in the fraction of near infrared that is transmitted transversely through the retina in an eyecup slice is monitored during stimulation by visible, green light. Near-infrared transmission increases both at the onset and at the termination of a step stimulus. This “ON/OFF” response is maximal in the neural retina. Sodium L-aspartate, which blocks the light-evoked activity of post-photoreceptor neurons, abolishes the “ON/OFF” response. L-AP4, used as a selective blocker of the ON channel, reduces the “ON” component and has little or no effect on the “OFF” component. The “ON” and “OFF” processes observed optically are distinct from those that generate the b– and d–waves of the electroretinogram, and the “ON” and “OFF” components may be superior to the b– and d–waves as indicators of ON and OFF channel activity. The optical method is almost as simple as electroretinography and has the advantage that responses can be spatially localized with ease.

Spatial summation curves were determined in the circumscribed visual-field defects of five patients with blindsight. Under light-adapted conditions that favor the color-opponent system, increment thresholds for white and red targets presented on a white background were measured as a function of stimulus size which ranged from 9–110 min arc. In both normal and defective hemifields, summation was linear for the red stimuli. In contrast, the curves measured with the white stimuli showed some nonlinearity in the normal hemifield, and a pronounced eccentricity-dependent notch in the field defect. The results indicate that the neurons mediating sensitivity differ in their summation properties for wavelength and intensity information.

A perfusion system was used to monitor the release of [3H]-GABA from isolated retinas of Xenopus laevis. Measurable release was stimulated by glycine at concentrations as low as 200 μM. Glycine-stimulated release was blocked by strychnine, and was not reduced in “calcium-free” Ringer's solution (0 Ca2+/20 mM Mg2+). Glutamate also stimulated calcium-independent release, using concentrations as low as 100 μM. In contrast, release stimulated by 25 mM potassium was reduced by 80% in calcium-free medium.

In most experiments, agonists were applied in six consecutive 4-mm pulses separated by 10-mm washes with Ringer's solution. Under these conditions, the release stimulated by 0.5 mM glutamate or 25 mM potassium decreased by at least 50% from the first to the second pulse, and then gradually decreased with successive applications. In contrast, the response to 0.5 mM glycine at first increased and then only gradually decreased with successive pulses. These patterns of response to different agonists were similar in calcium-free medium.

Somatostatin (—14 or —28) also stimulated release, and this effect was inhibited by AOAA, an inhibitor of GABA degradation. In the presence of AOAA, somatostatin had little effect, except at high concentrations of somatostatin (5 μM), which increased both basal and glycine-stimulated release. In contrast to somatostatin, glycine-stimulated release was much larger in the presence of AOAA.

Autoradiography was used to investigate which cell types released [3H]-GABA under our conditions. Autoradiograms showed that horizontal cells and a population of apparent “off” bipolar cells were well-labeled by [3H]-GABA high-affinity uptake. In addition, light labeling was seen over numerous amacrine cells. After application of glycine, glutamate, or potassium, there was a decrease in label density over horizontal cells.

Cat-301 is a monoclonal antibody which recognizes a cell surface associated antigen of selected neurons in the central nervous system (CNS). In the visual system, cat-301 selectively labels Y-like cells in several visual structures, including portions of the lateral geniculate nucleus complex and visual cortex. The cat superior colliculus (SC) also receives Y input and contains cells driven by Y input which are selectively distributed in the deep superficial gray and deeper laminae. If cat-301 is selective to the Y-cell system in SC, labeled cells should be restricted to those laminae. To test this hypothesis, we have examined quantitatively the laminar distribution, percentage, size, and morphology of cells in SC labeled by the cat-301 antibody. Cat-301 labeled a variety of cells in the cat SC. Labeled cells were found within the deep portion of the superficial gray layer (6.6%), optic layer (27.6%), intermediate gray layer (26.9%), and the deep gray and white layers (38.5%). By contrast, only 2 of 667 labeled cells (0.3%) were found within that part of the upper superficial gray layer innervated exclusively by W input and thought to contain only W-driven cells. When considered as a percentage of the total cell population, cat-301 labeled cells represented less than 3% of cells in the superficial gray layer and approximately 15% in the deeper layers. Neurons labeled by cat-301 were all of medium to large size (mean average diameter = 33.3μm; range = 15–84μm) and included vertical fusiform and stellate cells in the upper layers and the very large neurons found in the intermediate gray and deeper layers. These results provide further evidence that the cat-301 antibody selectively recognizes the Y channel of the cat visual system.

The vast majority of neurons of the rat medial terminal nucleus (MTN) project to the nucleus of the optic tract (NOT), but the MTN also projects to a lesser degree upon a number of other brainstem nuclei controlling optokinetic nystagmus. Because of the diversity of targets of the MTN, it is possible that individual neurons have branched axons that project to two or more brainstem nuclei. The possibility that axons of MTN-NOT neurons collateralize to innervate other MTN targets is examined in the rat with the fluorescent, double-labeling, retrograde tracer technique. Fluoro-Gold was injected into the NOT while Fast Blue was simultaneously injected into each of five other known targets of the MTN: the supraoculomotor-periaqueductal gray; the dorsal cap of the inferior olive; the visual tegmental relay zone; the dorsolateral nucleus of the basal pons; and the superior/lateral vestibular nuclei. Brainstem sections were processed for fluorescence microscopy and the MTN was examined for single- and double-labeled neurons. Results show that virtually all neurons of the MTN (>97.5%), together with neurons in the visual tegmental relay zone immediately surrounding the MTNd, are single-labeled in all paired injections involving the NOT and the other target nuclei. It was found that about 69% of MTN neurons project exclusively to the NOT, 5–10% project to each one of the other nuclei, and 3% of MTN neurons project to more than one target. Based upon cell counts from the fluorescent material, and previous analysis of Nissl-stained serial sections, the findings show that virtually all MTN neurons are projection neurons. It was concluded that the MTN is comprised of independent projection systems, possibly involved in different aspects of generating optokinetic nystagmus.

Cells projecting into the commissure of the cat's superior colliculus were identified during extracellular recording by antidromic activation. Electrical stimulation of the ipsilateral optic tract evoked action potentials in the majority of commissural neurons. Response latencies of 1.4 ± 0.5 ms (mean ± S.D.) in a few cells indicated that some commissural neurons receive direct input from the axons of retinal Y–cells. Most commissural cells responded 5.9 ± 1.9 ms (mean ± S.D.) following optic tract shock, implying that the responsible pathway was composed of more slowly conducting axons or did not proceed directly to the colliculus. Results of previous studies of retinal inputs to cells in the deep tectal layers suggest that the later responses were mediated by an indirect Y pathway through the visual cortex.

Physiological studies of intracellular messengers frequently employ intracellular injections of large molecules that either monitor or modulate the metabolism of the messenger cascade. Injected molecules have unknown mobility in the cytosol and unknown accessibility to various cytosolic compartments, including those postulated to be traversed by intracellular messenger molecules. In order to determine whether injected molecules have access to the confined spaces through which messenger molecules must diffuse, we injected 5-nm colloidal gold or horseradish peroxidase, or both, into Limulus ventral photoreceptors. Injections were made by applying pressure pulses to the back of an intracellular micropipette that also monitored membrane voltage. The tissue was fixed at varying times after injection and processed for electron microscopy by conventional techniques. Cells fixed 1–3 min after injection contained HRP reaction product only in the cell body. HRP reaction product was found at varying distances down axons in direct relation to the interval between injection and fixation. Colloidal gold particles were found throughout the cell body but not in axons of tissue fixed 1–3 min after injection. Both HRP reaction product and 5-nm colloidal gold particles were observed within the microvillar projections of internal and external rhabdomere, as well as within the extracisternal spaces of endoplasmic reticulum. We conclude that large molecules injected from an intracellular micropipette into an arbitrary locus of ventral photoreceptor cells have access to all of the presumed sites of the phototransduction cascade.

We studied the development of sensitivity to complex motion using plaid patterns. We hypothesized, based on neurophysiological data showing a dearth of pattern direction–selective (PDS) cells in area medial temporal (MT) of infant macaques, that sensitivity to pattern motion would develop later than other forms of global motion sensitivity. We tested 10 macaque monkeys (Macaca nemestrina) ranging in age from 7 weeks to 109–160 weeks (adult). The monkeys discriminated horizontal from vertical pattern motion; sensitivity for one-dimensional (1D) direction discrimination and detection were tested as control tasks. The results show that pattern motion discrimination ability develops relatively late, between 10 and 18 weeks, while performance on the 1D control tasks was excellent at the earliest test ages. Plaid discrimination performance depends on both the speed and spatial scale of the underlying patterns. However, development is not limited by contrast sensitivity. These results support the idea that pattern motion perception depends on a different mechanism than other forms of global motion perception and are consistent with the idea that the representation of PDS neurons in MT may limit the development of complex motion perception.

There is common consensus now that color-defined motion can be perceived by the human visual system. For global motion integration tasks based on isoluminant random dot kinematograms conflicting evidence exists, whether observers can (Ruppertsberg et al., 2003) or cannot (Bilodeau & Faubert, 1999) extract a common motion direction for stimuli modulated along the isoluminant red-green axis. Here we report conditions, in which S-cones contribute to chromatic global motion processing. When the display included extra-foveal regions, the individual elements were large (∼0.3°) and the displacement was large (∼1°), stimuli modulated along the yellowish-violet axis proved to be effective in a global motion task. The color contrast thresholds for detection for both color axes were well below the contrasts required for global motion integration, and therefore the discrimination-to-detection ratio was >1. We conclude that there is significant S-cone input to chromatic global motion processing and the extraction of global motion is not mediated by the same mechanism as simple detection. Whether the koniocellular or the magnocellular pathway is involved in transmitting S-cone signals is a topic of current debate (Chatterjee & Callaway, 2002).

Immunocytochemical localization was carried out for five isoforms of protein kinase C (PKC) in the cat retina. In common with other mammalian species, PKCα was found in rod bipolar cells. Staining was also seen in a small population of cone bipolar cells with axon terminals ramifying near the middle of the inner plexiform layer (IPL). PKCβI was localized to rod bipolar cells, one class of cone bipolar cell, and numerous amacrine and displaced amacrine cells. Staining for PKCβII was seen in three types of cone bipolar cells as well as in amacrine and ganglion cells. Immunoreactivity for both PKCε and PKCζ was found in rod bipolar cells; PKCε was also seen in a population of cone bipolar cells and a few amacrine and ganglion cells whereas PKCζ was found in all ganglion cells. Double-label immunofluorescence studies showed that dendrites of the two PKCβII-positive OFF-cone bipolar cells exhibit immmunoreactivity for the kainate-selective glutamate receptor GluR5. The third PKCβII cone bipolar is an ON-type cell and did not stain for GluR5. The retinal distribution of these isoforms of PKC is consistent with a role in modulation of various aspects of neurotransmission including synaptic vesicle release and regulation of receptor molecules.

SM, a 21-year-old female, presents an extensive central scotoma (30 deg) with dense absolute scotoma (visual acuity = 10/100) in the macular area (10 deg) due to Stargardt's disease. We provide behavioral evidence of cortical plastic reorganization since the patient could perform several visual tasks with her poor-vision eyes better than controls, although high spatial frequency sensitivity and visual acuity are severely impaired. Between 2.5-deg and 12-deg eccentricities, SM presented (1) normal acuity for crowded letters, provided stimulus size is above acuity thresholds for single letters; (2) a two-fold sensitivity increase (d-prime) with respect to controls in a simple search task; and (3) largely above-threshold performance in a lexical decision task carried out randomly by controls. SM's hyper-vision may reflect a long-term sensory gain specific for unimpaired low spatial-frequency mechanisms, which may result from modifications in response properties due to practice-dependent changes in excitatory/inhibitory intracortical connections.

The ability of color-deficient observers to discriminate between illuminant changes and surface-reflectance changes in a scene was tested with natural and Munsell reflectance spectra. To avoid the confounding effects of spatial structure, stimuli were simulations of Mondrian-like colored patterns, presented on a computer-controlled color monitor. Protanopes performed less well than normal trichromats, regardless of the type of reflectance spectra, but they were least disadvantaged with patterns comprising reflectance spectra drawn from urban and rural scenes, more characteristic of the natural environment.

Phosducin (Pd) is a 28-kD phosphoprotein whose expression in retina appears limited to photoreceptor cells. Pd binds to the β,γ subunits of transducin (Gt). Their binding affinity is markedly diminished by Pd phosphorylation. While Pd has long been regarded as a candidate for the regulation of Gt, the molecular details of Pd function remain unclear. This gap in understanding is due in part to a lack of precise information concerning the total amount and subcellular localization of rod Pd. While earlier studies suggested that Pd was a rod outer segment (ROS) protein, recent findings have demonstrated that Pd is distributed throughout the rod. In this report, the subcellular distribution and amounts of rat Pd are quantified with immunogold electron microscopy. After light or dark adaptation, retinal tissues were fixed in situ and prepared for ultrathin sectioning and immunogold labeling. Pd concentrations were analyzed over the entire length of the rod. The highest Pd labeling densities were found in the rod synapse. Less intense Pd staining was observed in the ellipsoid and myoid regions, while minimal labeling densities were found in the ROS and the rod nucleus. In contrast with rod Gt, no evidence was found for light-dependent movement of Pd between inner and outer segments. There is a relative paucity of Pd in the ROS as compared with the large amounts of Gt found there. This does not support the earlier idea that Pd could modulate Gt activity by controlling its concentration. On the other hand, the presence of Pd in the nucleus is consistent with its possible role as a regulator of transcription. The functions of Pd in the ellipsoid and myoid regions remain unclear. The highest concentration of Pd was found at the rod synapse, consistent with a suggested role for Pd in the regulation of synaptic function.

We measured the latency of 35 cells from V1 in two rhesus monkeys, to dynamic random dot stimuli monocular and binocularly presented. Mean latencies after non-dominant eye stimulation (97.9 ms) were longer than those for dominant eye (78.2 ms) and binocular (70.7 ms) stimulation. Differences between latencies for dominant eye and binocular stimulation were not statistically significant. For dominant eye, there was a significant statistical correlation between dominance strength and latency (R = −0.36; p = 0.03). We failed to find significant statistical differences between latencies for cells with temporal and nasal dominant receptive-field. We conclude that, in V1, the response latency is largely determined by the dominant eye, whereas interocular interactions do not seem to play a relevant role regarding response latency.

Antisera directed against hyperpolarization-activated, cyclic nucleotide–sensitive (HCN) channels bind to somata in the ganglion cell layer of rat and rabbit retinas, and mRNA for different HCN channel isoforms has been detected in the ganglion cell layer of mouse retina. However, previous studies neither provided evidence that any of the somata are ganglion cells (as opposed to displaced amacrine cells) nor quantified these cells. We therefore tested whether isoform-specific anti-HCN channel antisera bind to ganglion cells labeled by retrograde transport of fluorophore-coupled dextran. In flat-mounted adult rat retinas, the number of dextran-backfilled ganglion cells agreed with cell densities reported in previous studies, and anti-HCN4 antisera bound to the somata of approximately 40% of these cells. The diameter of these somata ranged from 7 to 30 μm. Consistent with localization to cell membranes, the immunoreactivity formed a thin line that circumscribed individual somata. Optic fiber layer axon fascicles, and the proximal dendrites of some ganglion cells, also displayed binding of anti-HCN4 antisera. These results suggest that the response of some mammalian retinal ganglion cells to hyperpolarization may be modulated by changes in intracellular cAMP levels, and could thus be more complex than expected from previous voltage and current recordings.

Previous studies have shown that multiple sclerosis (MS) affects the visual system, mainly by reducing contrast sensitivity (CS), a function that can be assessed by measuring contrast sensitivity function (CSF). To this end, we measured both the CSF for sine-wave gratings and angular frequency stimuli with 20 participants aged between 21 and 44 years, of both genders, with normal or corrected to normal visual acuity. Of these 20 participants, there were 10 volunteers with clinically defined MS of the relapsing–remitting clinical form, with no history of optic neuritis (ON), as well as 10 healthy volunteers who served as the control group (CG). We used a forced-choice detection paradigm. The results showed reduced CS to both classes of stimuli. Differences were found for sine-wave gratings at spatial frequencies of 0.5, 1.25, and 2.5 cycles per degree (cpd) (P < 0.002) and for angular frequency stimuli of 4, 24, and 48 cycles/360° (P < 0.05). On the one hand, comparing the maxima of the respective CSFs, the CS to angular frequency stimuli (24 cycles/360°) was 1.61-fold higher than that of the CS to vertical sine-wave gratings (4.0 cpd) in the CG; for the MS group, these values were 1.55-fold higher. On the other hand, CS in the MS group attained only 75% for 24 cycles/360° and 78% for 4.0 cpd of the 100% CS estimates found for the CG at the peak frequencies. These findings suggest that MS affects the visual system, mostly at its maximum contrast sensitivities. Also, since angular frequencies and sine-wave gratings operate at distinct levels of contrast in the visual system, MS seems to affect CS at both high and low levels of contrast.

Vision begins with photoisomerization of 11-cis retinal to the all-trans conformation within the chromophore-binding pocket of opsin, leading to activation of a biochemical cascade. Release of all-trans retinal from the binding pocket curtails but does not fully quench the ability of opsin to activate transducin. All-trans retinal and some other analogs, such as β-ionone, enhance opsin’s activity, presumably on binding the empty chromophore-binding pocket. By recording from isolated salamander photoreceptors and from patches of rod outer segment membrane, we now show that high concentrations of β-ionone suppressed circulating current in dark-adapted green-sensitive rods by inhibiting the cyclic nucleotide-gated channels. There were also decreases in circulating current and flash sensitivity, and accelerated flash response kinetics in dark-adapted blue-sensitive (BS) rods and cones, and in ultraviolet-sensitive cones, at concentrations too low to inhibit the channels. These effects persisted in BS rods even after incubation with 9-cis retinal to ensure complete regeneration of their visual pigment. After long exposures to high concentrations of β-ionone, recovery was incomplete unless 9-cis retinal was given, indicating that visual pigment had been bleached. Therefore, we propose that β-ionone activates and bleaches some types of visual pigments, mimicking the effects of light.

The pond turtle (Trachemys scripta elegans) exhibits a notably sluggish pupillary light reflex (PLR), with pupil constriction developing over several minutes following light onset. In the present study, we examined the dynamics of the efferent branch of the reflex in vitro using preparations consisting of either the isolated head or the enucleated eye. Stimulation of the oculomotor nerve (nIII) using 100-Hz current trains resulted in a maximal pupil constriction of 17.4% compared to 27.1% observed in the intact animal in response to light. When current amplitude was systematically increased from 1 to 400 μA, mean response latency decreased from 64 to 45 ms, but this change was not statistically significant. Hill equations fitted to these responses indicated a current threshold of 3.8 μA. Stimulation using single pulses evoked a smaller constriction (3.8%) with response latencies and threshold similar to that obtained using train stimulation. The response evoked by postganglionic stimulation of the ciliary nerve using 100-Hz trains was largely indistinguishable from that of train stimulation of nIII. However, application of single-pulse stimulation postganglionically resulted in smaller pupil constriction at all current levels relative to that of nIII stimulation, suggesting that there is amplification of efferent drive at the ganglion. Time constants for constrictions ranged from 88 to 154 ms with relaxations occurring more slowly at 174–361 ms. These values for timing from in vitro are much faster than the time constant 1.66 min obtained for the light response in the intact animal. The rapid dynamics of pupil constriction observed here suggest that the slow PLR of the turtle observed in vivo is not due to limitations of the efferent pathway. Rather, the sluggish response probably results from photoreceptive mechanisms or central processing.

The number, dendritic morphology, and retinal distribution of displaced ganglion cells were studied in two anuran species, Xenopus laevis and Bufo marinus. Horseradish peroxidase or cobaltic lysine complex was applied to the cut end of the optic nerve, and the size, shape, and retinal position of retrogradely filled ganglion cells displaced into the inner nuclear layer were determined in retinal wholemount and sectioned material. Approximately 1% of ganglion cells in Xenopus and 0.1% in Bufo were found to be displaced. In both species, many of the previously described orthotopic ganglion cell types (Straznicky & Straznicky, 1988; Straznicky et al., 1990) were present among displaced ganglion cells. In Xenopus more displaced ganglion cells were found in the retinal periphery than in the retinal center, and they formed 3 or 4 distinct bands around the optic nerve head. In Bufo the incidence of displaced ganglion cells was higher along the visual streak than in the dorsal and ventral peripheral retina. These results indicate that the distribution of displaced ganglion cells approximates the retinal distribution of orthotopic ganglion cells. One of the likely mechanisms to account for this developmental paradox may be that the formation of the inner plexiform layer, adjacent to the ciliary margin, acts as a mechanical barrier by preventing the entry of some of the late developing ganglion cells into the ganglion cell layer.

Long- and medium-wavelength cones in the turtle retina participate in complex neural interactions. They are coupled via excitatory pathways to other cones and receive negative feedback inputs from luminosity-type horizontal cells. Little information has been collected on the S- (short-wavelength or blue) cones because they are scarce in the turtle retina and of smaller dimensions compared to the other cone types.

In this paper, flash sensitivity action spectra and photoresponses of seven turtle S-cones were measured in the dark-adapted state and during chromatic background illuminations. The desensitizing action of monochromatic background lights was not uniform across the visible spectrum. A red background was most effective in desensitizing the S-cones to long-wavelength stimuli while a blue background light produced its strongest action on the photoresponses elicited by short-wavelength stimuli. The effects of chromatic adaptation on the S-cone action spectrum and on the kinetics of the small-amplitude photoresponses suggested that the S-cones in the turtle retina were involved in complex neural interactions. These included excitatory inputs probably originating in neighboring L-cones and inhibitory long-wavelength inputs probably mediated by L-type horizontal cells.