Smooth pursuit eye movements allow primates to keep gaze pointed at small objects moving across stationary surroundings. In monkeys trained to track a small moving target, we have injected brief perturbations of target motion under different initial conditions as probes to read out the state of the visuo-motor pathways that guide pursuit. A large eye movement response was evoked if the perturbation was applied to a moving target the monkey was tracking. A small response was evoked if the same perturbation was applied to a stationary target the monkey was fixating. The gain of the response to the perturbation increased as a function of the initial speed of target motion and as a function of the interval from the onset of target motion to the time of the perturbation. The response to the perturbation also was direction selective. Gain was largest if the perturbation was along the axis of ongoing target motion and smallest if the perturbation was orthogonal to the axis of target motion. We suggest that two parallel sets of visual motion pathways through the extrastriate visual cortex may mediate, respectively, the visuo-motor processing for pursuit and the modulation of the gain of transmission through those pathways.

Axon diameters of retinal ganglion cells were measured from electron micrographs of the nerve fiber layer of the cat. Three adult retinae were examined which had mean axonal diameters of 1.18 ± 0.86 (n = 5553), 1.12 ± 0.79 (n = 7265), and 1.47 ±1.11 μm (n = 10,867). Cumulative histograms from several locations adjacent to the optic disc were unimodal (modal peaks: 0.6–0.8 μm). This unimodal distribution, however, did not reflect the regional differences in axonal diameters found throughout the retina. In many locations, especially those related to axons of the temporal retina, axon diameter distributions were clearly bimodal or even trimodal (modal peaks: 0.6–0.8, 1.4–2.1, and 3.3 μm). Measurements from one retina indicated that the mean diameters of axons arising from the area centralis and visual streak (0.94 ± 0.63 and 0.98 ± 0.68, respectively) were not significantly different from each other; however, when compared to other areas around the optic disc, the percentage of fibers with diameters between 1.5–2.0 μm was highest in the sample adjacent to the area centralis. Axons temporal to the optic disc were found to be on average larger than those nasal to the optic disc; similarly superior axons were larger than inferior axons. Axonal distributions at the retinal periphery were found to be significantly different from those at the optic disc (p ≤ 0.05) and contained a higher percentage of medium-sized axons and fewer small axons. In each of the three retinae the proportions small, medium, and large axons were respectively γ: 46; 47; 48, β: 50; 49; 48, and α: 4; 4; 4; regional differences in the proportions of each axonal class are compared to previously published ganglion cell density maps. Differences between axonal bundles within each sample location were not significantly different; however, in one retina axons in the scleral half of the fiber layer were significantly larger (P ≤ 0.01) than axons in the vitreal half of the nerve fiber layer adjacent to the optic disc. When compared to the axonal diameter distributions found within the optic nerve (Cottee et al., 1991) and optic tract (Reese et al., 1991), our data indicates that the diameter of retinal axons may increase by up to 30% along the length of the visual pathway.

Anterograde transport of cholera toxin subunit B (CTb) was used to study the retinal projections in birds, with an emphasis on retinohypothalamic connections. Pigeons (Columba livia) were deeply anesthetized and received unilateral intraocular injections of CTb. In addition to known contralateral retinorecipient regions, CTb-immunoreactive fibers and presumptive terminals were found in several ipsilateral regions, such as the nucleus of the basal optic root, ventral lateral geniculate nucleus, intergeniculate leaflet, nucleus lateralis anterior, area pretectalis, and nucleus pretectalis diffusus. In the hypothalamus, CTb-immunoreactive fibers were observed in at least two contralateral cell groups, a medial hypothalamic retinorecipient nucleus, and a lateral hypothalamic retinorecipient nucleus. To compare retinorecipient hypothalamic nuclei in pigeons with the mammalian suprachiasmatic nucleus, double-label experiments were conducted to study the existence of neurophysin-like immunoreactivity in the retinorecipient avian hypothalamus. The results showed that only cell bodies in the medial hypothalamic nucleus contained neurophysin-like immunoreactivity. The results demonstrate CTb to be a sensitive anterograde tracer and provide further anatomical information on the avian equivalent of the mammalian suprachiasmatic nucleus.

Naturally occurring cell death has been hypothesized to sculpt various features of the organization of the mature visual pathways, including the recent proposal that the selective elimination of ganglion cells in the temporal retina shapes the formation of decussation patterns. Through a class-specific interocular competition, ganglion cells in the two temporal hemiretinae are selectively lost to produce the decussation patterns characteristic of each individual cell class (Leventhal et al., 1988). The present study has tested this hypothesis by asking whether the removal of one retina in newborn ferrets, which should disrupt binocular interactions at the level of the terminals, alters the decussation pattern of the alpha cells, a cell class that is entirely decussating in the normal adult ferret. Enucleation on the day of birth was found to increase the uncrossed projection by ≈50%, but not a single uncrossed alpha cell was found in the temporal retina. Either alpha cells never project ipsilaterally during development, or if they do, they cannot be rescued by early enucleation. While naturally occurring cell death plays many roles during development, creating the decussation pattern of the ferreth's alpha cell class via a binocular competition at the level of the targets is unlikely to be one of them.

We have studied the responses of MT neurons to moving gratings, spatially modulated in luminance and chromaticity. Most MT neurons responded briskly and with high contrast sensitivity to targets whose luminance was modulated, with or without added chromatic contrast. When luminance modulation was removed and only chromatic stimulation was used, the responses of all MT neurons were attenuated. Most were completely unresponsive to stimulation with targets whose modulation fell within a “null” plane in color space; these null planes varied from neuron to neuron, but all lay close to the plane of constant photometric luminance. For about a third of the neurons, there was no color direction in which responses were completely abolished; almost all of these neurons had a definite minimum response for chromatic modulation near the isoluminant plane. MT neurons that responded to isoluminant targets did so inconsistently and with poor contrast sensitivity, so that only intensely modulated targets were effective. Whereas the best thresholds of MT neurons for luminance targets are close to behavioral contrast threshold, the thresholds for isoluminant targets lie considerably above behavioral contrast threshold. Therefore, although some MT neurons do give responses to isoluminant targets, they are unlikely to be the source of the chromatic motion signals revealed behaviorally.

An important issue in understanding the function of primary visual cortex in the macaque monkey is how the several efferent neuron groups projecting to extrastriate cortex acquire their different response properties. To assist our understanding of this issue, we have compared the anatomical distribution of VI intrinsic relays that carry information derived from magno- (M) and parvocellular (P) divisions of the dorsal lateral geniculate nucleus between thalamic recipient neurons and interareal efferent neuron groups within area VI. We used small, iontophoretic injections of biocytin placed in individual cortical laminae of area VI to trace orthograde and retrograde inter- and intralaminar projections. In either the same or adjacent sections, the tissue was reacted for cytochrome oxidase (CO), which provides important landmarks for different efferent neuron populations located in CO rich blobs and CO poor interblobs in laminae ⅔, as well as defining clear boundaries for the populations of efferent neurons in laminae 4A and 4B. This study shows that the interblobs, but not the blobs, receive direct input from thalamic recipient 4C neurons; the interblobs receive relays from mid 4C neurons (believed to receive convergent M and P inputs), while blobs receive indirect inputs from either M or P (or both) pathways through layers 4B (which receives M relays from layer 4Cα) and 4A (which receives P relays directly from the thalamus as well as from layer 4Cβ). The property of orientation selectivity, most prominent in the interblob regions and in layer 4B, may have a common origin from oriented lateral projections made by mid 4C spiny stellate neurons. While layer 4B efferents may emphasize M characteristics and layer 4A efferents emphasize P characteristics, the dendrites of their constituent pyramidal neurons may provide anatomical access to the other channel since both blob and interblob regions in layers ⅔ have anatomical access to M and P driven relays, despite functional differences in the way these properties may be expressed in the two compartments.

Psychophysical studies have documented that many observers show lower thresholds for rapid-off than for rapid-on sawtooth luminance modulation. This finding, together with physiological findings from chromatically opponent ganglion cells of the macaque monkey, prompted a search for a similar bias in psychophysical detection of chromatic increments and decrements of light. Using a luminance pedestal in conjunction with a luminance background to favor detection by chromatic mechanisms, we measured spectral sensitivity for rapid-on and rapid-off sawtooth stimuli presented spatially coextensive with the pedestal. There were two different pedestal chromaticities: one broadband, and the second composed only of long-wavelength light to enhance short-wavelength-sensitive, cone-mediated detection. Spectral-sensitivity measurements for different wavelength stimuli revealed no systematic differences across the visible spectrum as a function of sawtooth waveform polarity or pedestal chromaticity. Similarly, temporal contrast-sensitivity functions for hetero-chromatically modulated red-green sawtooth stimuli did not reveal an asymmetry in sensitivity for rapid-red and rapid-green chromatic change. Some of the observers showed a higher sensitivity for luminance modulated rapid-off sawtooth stimuli, as also noted in previous studies. This asymmetry was not found when a white luminance pedestal and background was used. These results suggest that the cone inputs to chromatically opponent ON- and OFF-center cells are sufficiently balanced to provide equivalent psychophysical thresholds for chromatic increments and decrements of light.

Retinal horizontal cells of four rodent species, rat, mouse, gerbil, and guinea pig were examined to determine whether they conform to the basic pattern of two horizontal cell types found in other mammalian orders. Intracellular injections of Lucifer-Yellow were made to reveal the morphologies of individual cells. Immunocytochemistry with antisera against the calcium-binding proteins calbindin D-28k and parvalbumin was used to assess population densities and mosaics.

Lucifer-Yellow injections showed axonless A-type and axon-bearing B-type horizontal cells in guinea pig, but revealed only B-type cells in rat and gerbil retinae. Calbindin immunocytochemistry labeled the A-and B-type populations in guinea pig, but only a homogeneous regular mosaic of cells with B-type features in rat, mouse, and gerbil. All calbindin-immunoreactive horizontal cells in the latter species were also parvalbumin-immunoreactive; comparison with Nissl-stained retinae showed that both antisera label all of the horizontal cells. Taken together, the data from cell injections and the population studies provide strong evidence that rat, mouse, and gerbil retinae have only one type of horizontal cell, the axon-bearing B-type, where as the guinea pig has both A-and B-type cells. Thus, at least three members of the family Muridae differ from other rodents and deviate from the proposed mammalian scheme of horizontal cell types.

The absence of A-type cells is apparently not linked to any peculiarities in the photoreceptor populations, and there is no consistent match between the topographic distributions of the horizontal cells and those of the cone photoreceptors or ganglion cells across the four rodent species. However, the cone to horizontal cell ratio is rather similar in the species with and without A-type cells.

Existing models of the primate photopic electroretinogram (ERG) attribute the light-adapted b–wave to activity of depolarizing bipolar cells (DBCs), mediated through a release of potassium that is monitored by Müller cells. However, possible ERG contributions from OFF-bipolar cells (HBCs) and horizontal cells (HzCs) have not been explored. We examined the contribution of these hyperpolarizing second-order retinal cells to the photopic ERG of monkey by applying glutamate analogs to suppress photoreceptor transmission selectively to HBC/HzCs vs. DBCs.

ERGs of Macaca monkeys were recorded at the cornea before and after intravitreal injection of drugs. Photopic responses were elicited by bright 200–220 ms flashes on a steady background of 3.3 log scotopic troland to suppress rod ERG components.

2–amino-4–phosphonobutyric acid (APB), which blocks DBC light responses, abolished the photopic b–wave and indicated that DBC activity is requisite for photopic b–wave production.

However, applying cis–2,3–piperidine dicarboxylic acid (PDA) and kynurenic acid (KYN), to suppress HBCs/HzCs and third-order neurons, revealed a novel ERG response that was entirely positive and was sustained for the duration of the flash. The normally phasic b–wave was subsumed into this new response. Applying n–methyl-dl-aspartate (NMA) did not replicate the PDA+KYN effect, indicating that third-order retinal cells are not involved. This suggests that HBC/HzC activity is critical for shaping the phasic b–wave.

Components attributable to depolarizing vs. hyperpolarizing cells were separated by subtracting waveforms after each drug from responses immediately before. This analysis indicated that DBCs and HBC/HzCs each can produce large but opposing field potentials that nearly cancel and that normally leave only the residual phasic b–wave response in the photopic ERG.

Latency of the DBC component was 5–9 ms slower than the HBC/HzC component. However, once activated, the DBC component had a steeper slope. This resembles properties known for the two types of cone synapses in lower species, in which the sign-preserving HBC/HzC synapse has faster kinetics but probably lower gain than the slower sign-inverting G-protein coupled DBC synapse.

A human patient with “unilateral cone dystrophy” was found to have a positive and sustained ERG that mimicked the monkey ERG after PDA+KYN, indicating that these novel positive photopic responses can occur naturally even without drug application.

These results demonstrate that hyperpolarizing second-order neurons are important for the primate photopic ERG. A “Push-Pull Model” is proposed in which DBC activity is requisite for b–wave production but in which HBC/HzC activity limits the amplitude and controls the shape of the primate photopic b–wave.

Retinal bipolar cells exhibit a center-surround antagonistic receptive field to a light stimulus (Werblin & Dowling, 1969; Kaneko, 1970), and thus constitute an early stage of spatial information processing. We injected Lucifer Yellow and a small biotinylated tracer, biocytin, into bipolar cells of the teleost retina to examine electrical coupling in these cells. Lucifer-Yellow coupling was observed in one of 55 stained bipolar cells; the coupling pattern was one injected bipolar cell and three surrounding cells. Biocytin coupling was observed in 16 of 55 stained bipolar cells, six of which were ON center and ten OFF center. Although biocytin usually coupled to three to six bipolar cells, some OFF-center bipolar cells showed strong coupling to more than 20 cells. The biocytin-coupled bipolar cells were morphologically homologous. Membrane appositions resembling gap junctions were found between dendrites and between axon terminals of neighboring bipolar cells.

In the strongest biocytin-coupled bipolar cells, the contacts between bipolar cells and cone photoreceptor cells were examined after reconstruction of the dendritic trees of five well-stained, serially sectioned OFF-center bipolar cells. Each of these bipolar cells was in contact with different numbers of cones: 11 to 20 for twin cones and two to four for single cones. This implies that, although these bipolar cells belong to the same category, the signal inputs differ among bipolar cells. Numerical simulation conducted on a hexagonal array network model demonstrated that the electrical coupling of bipolar cells can decrease the difference in input (≈80%) without causing significant loss of spatial resolution. Our results suggest that electrical coupling of bipolar cells has the advantage of decreasing the dispersion of input signals from cones, and permits bipolar cells of the same class to respond to light with similar properties.

Both A- and B-type horizontal cells in the rabbit retina were labeled by brief in vitro incubations of the isolated retina in the blue fluorescent dye 4,6–diamino-2–phenylindole. Intracellular injection of Lucifer Yellow into the somata revealed the morphology of the individual cells. Dye-coupling with Lucifer Yellow was seen only between A-type horizontal cells. By contrast, injection of the tracer Neurobiotin showed dye-coupling between both A- and B-type horizontal cells. There also appeared to be coupling between the axon terminals of B-type horizontal cells.

The extensive dye-coupling seen following injection of Neurobiotin into a single horizontal cell soma can be used to obtain population counts of each cell type. Staining of large numbers of each cell type across the retina showed that each type increased in number and declined in dendritic diameter as the visual streak was approached, such that relatively constant coverage across the retina was maintained. In the visual streak, A-type horizontal cells numbered 555 cells/mm2 and averaged 120 μm in diameter, compared to 1375 cells/mm2 and 100 μm for B-type horizontal cells. In the periphery, the A- and B-types numbered 250 cells/mm2 and 400 cells/mm2, respectively. The average diameters of the dendritic trees at these locations were 225 μm for the A-type and 175 μm for the B-type. Coverage across the retina averaged almost six for A-type horizontal cells and 8–10 for the B-type. A-type horizontal cells in the visual streak whose elliptical dendritic fields were shown by Bloomfield (1992) to correlate physiologically with orientation bias were shown to be dye-coupled to cells with symmetrical dendritic fields.

The immunocytochemical labeling of arrestin (S-antigen) in photoreceptors of the ovine retina was examined following incubation of the retina with hydroxylamine (NH2OH), an agent known to inhibit the phosphorylation of photoactivated rhodopsin. Intact, isolated retinas bathed in medium containing 20 mM NH2OH, or in control medium lacking NH2OH, were maintained in darkness or exposed to bright light for 3 min (dark-adapted and light-adapted conditions, respectively); further incubated in darkness for 10 min; and then fixed and prepared for cryosectioning. Cryosections were incubated with anti-S-antigen monoclonal antibody MAb A2G5; with secondary antibodies that were conjugated with horseradish peroxidase; and with either 3–amino-9–ethyl carbazole or diaminobenzidine as chromogen. Anti-arrestin labeling in cryosections was then analyzed densitometrically using a light-microscopic image processing system. In dark-adapted control retinas, labeling density of the photoreceptor outer segment (OS) layer (0.061 ± 0.004; average ± S.e.m.) was less than that of the inner segment (IS) layer (0.138 ± 0.011). In light-adapted control retinas, OS labeling density (0.139 ± 0.007) exceeded IS labeling density (0.095 ± 0.005). Incubation with NH2OH eliminated this light-dependent increase in labeling of the OS relative to that of the IS, i.e. eliminated the increase in relative OS/IS labeling. Densities of labeling were 0.110 ± 0.006 (OS) and 0.183 ± 0.006 (IS) in NH2OH-treated dark-adapted retinas vs. 0.078 ± 0.004 (OS) and 0.182 ± 0.008 (IS) in NH2OH-treated light-adapted retinas. Anti-arrestin labeling was also examined in retinas that were exposed to 3 min or 13 min of bright light and then immediately fixed. Among retinas incubated in the absence of NH2OH, an increase in OS/IS labeling density was evident after 3 min of illumination, and retinas illuminated for 13 min exhibited an even larger increase in OS/IS labeling. An increase in OS/IS labeling was also exhibited by NH2OH-treated retinas that had been illuminated for 3 min; by comparison with dark-adapted NH2OH-treated controls (average value of OS/IS labeling: 0.60), OS/IS labeling in these illuminated retinas was 0.97. However, OS/IS labeling in NH2OH-treated retinas that had been illuminated for 13 min (average value: 0.35) was lower than that of the dark-adapted controls. The results indicate that, within intact rods, NH2OH inhibits the light-dependent increase in OS/IS anti-arrestin labeling that is ordinarily expressed at long times (~10 min) after major bleaching of the visual pigment. Among the possible bases for the effect of NH2OH are a reduction in the driving force for the movement of arrestin from the inner to the outer segment and/or a facilitation of the degradation of arrestin in the outer segment.

Immunohistochemical and in situ hybridization techniques were used to localize the β4 subunit of the neuronal nicotinic acetylcholine receptors (nAChRs) in the ground squirrel retina. The β4 nAChR subunit was detected in both transverse and horizontal sections of the retina using a subunit-specific antiserum and the avidin-biotin complex technique. Two bands of labeled processes were seen in the inner plexiform layer, corresponding approximately to the laminae where the cholinergic cells arborize. Labeled cells were found in the ganglion cell layer and the inner third of the inner nuclear layer. The cells in the ganglion cell layer were medium- to large-sized and were frequently observed to give rise to axon-like processes. Most of the labeled neurons in the inner nuclear layer were small presumptive amacrine cells, but a few medium-to-large cells were also labeled. These could constitute a different class of amacrine cells or displaced ganglion cells. The latter possibility is supported by the existence of nAChR-containing displaced ganglion cells in the avian retina. In situ hybridization with a 35S-labeled cRNA probe revealed the expression of mRNA coding for the nAChR β4 subunit in the ganglion cell layer and the inner third of the inner nuclear layer. This finding confirmed the immunohistochemical data of the cellular localization of β4 nAChR subunit.

These results indicate that the β4 nAChR subunit is expressed by specific subtypes of neurons on the ground squirrel retina. As the expression of that particular nAChR subunit appears to be very limited in the brain, the present data suggest that the retina might represent a useful model to study the function of nAChRs containing the β4 subunit.

Extrastriate areas TEO and V4 have been associated with form and color vision. Area V4 has also been suggested to participate in processes concerned with attention, stimulus salience, and perceptual learning. In a continuing effort to elucidate the connectional interactions and microcircuitry of these areas, we describe in this report the pattern of feedback connections from TEO and V4. Connections were demonstrated by injections of the high-resolution anterograde tracers PHA-L or biocytin and further analyzed by reconstruction of 25 individual axons through serial sections. This analysis yielded several new results: (1) Both areas TEO and V4 have widespread feedback connections (defined by their preferential termination in layer 1 and avoidance of layer 4). From TEO, there are dense projections to area V4 and moderate ones to V2 and V1. From V4, there are dense projections to V2 and moderate ones to V3 and V1. (2) Terminal fields span large territories in area V1, up to 6.0 mm in the case of axons originating from TEO; up to 5.0 mm in the case of axons originating from V4. In V2, fields tend to be smaller, between 3.0–5.0 mm. (3) Many axons from TEO and some from V4 have terminations in both areas V1 and V2. (4) Because individual terminal clusters and segments are often larger than cytochrome oxidase compartments, especially in V1, we suggest they may not be correlated with this compartmental organization. These results are consistent with the hypothesis that feedback connections may contribute to processes other than perceptual discrimination.

We studied the effect of cooling the striate cortex on parvocellular lateral geniculate nucleus (PLGN) neurons in awake monkeys. Cooling the striate cortex produced both facilitation and inhibition of the responses of all neurons, depending on the stimulus presented. Cooling the striate cortex also altered the temporal distribution of spikes in the responses of PLGN neurons. Shannon's information measure revealed that cooling the striate cortex reduced the average stimulus-related information transmitted by all PLGN neurons. The reduction in transmitted information was associated with both facilitation and inhibition of the response. Cooling the striate cortex reduced the amount of information transmitted about all of the stimulus parameters tested: pattern, luminance, spatial contrast, and sequential contrast. The effect of cooling was nearly the same for codes based on the number of spikes in the response as for codes based on their temporal distribution. The reduction in transmitted information occurred because the differences among the responses to different stimuli (signal separation) were reduced, not because the variability of the responses to individual stimuli (noise) was increased. We conclude that one function of corticogeniculate feedback is to improve the ability of PLGN neurons to discriminate among stimuli by enhancing the differences among their responses.

In the δAsn20 Drosophila stock, the N-linked glycosylation site of opsin in Rl-6 receptors (Rhl) is absent. We used electroretinography (ERG), microspectrophotometry (MSP), and electron microscopy (EM) to quantify visual cell defects. Positive controls, w9, had wild type Rhl. MSP revealed minimal photopigment in δAsn20 for 6 days posteclosion; w9 had near normal visual pigment. ERG sensitivity and prolonged depolarizing afterpotential (PDA) were compared for δAsn20 and w9. δAsn20's Rl-6 function is decreased 100–fold at eclosion and diminishes until only R7/8 functions at 11 days. What little rhodopsin is routed to the rhabdomere functions. Morphometry showed smaller Rl-6 rhabdomeres in δAsn20 for 8 days posteclosion. Rhabdomeres in w9 were normal. A negative control, ninaE0117, a deletion of the Rhl gene, also has small rhabdomeres. δAsn20 and ninaE0117 lack the extreme rhabdomere elimination of ora (outer rhabdomeres absent), a nonsense mutant interrupting Rhl's coding sequence. δAsn20 and ora have surplus membrane while ninaE0117 does not. Freeze fracture reveals that δAsn20's rhabdomeric P-face particle count is as low as for vitamin A deprivation, consistent with an opsin defect. High particle density, organized into rows, is present in adjacent plasmalemma where surplus membrane accumulates. In summary, δAsn20 interferes with either synthesis, deployment, or maintenance of opsin.