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Functional analyses exist only for a few of the morphologically described primate ganglion cell types, and their correlates in other mammalian species remain elusive. Here, we recorded light responses of broad thorny cells in the whole-mounted macaque retina. They showed ON-OFF-center light responses that were strongly suppressed by stimulation of the receptive field surround. Spike responses were delayed compared with parasol ganglion cells and other ON-OFF cells, including recursive bistratified ganglion cells and A1 amacrine cells. The receptive field structure was shaped by direct excitatory synaptic input and strong presynaptic and postsynaptic inhibition in both ON and OFF pathways. The cells responded strongly to dark or bright stimuli moving either in or out of the receptive field, independent of the direction of motion. However, they did not show a maintained spike response either to a uniform background or to a drifting plaid pattern. These properties could be ideally suited for guiding movements involved in visual pursuit. The functional characteristics reported here permit the first direct cross-species comparison of putative homologous ganglion cell types. Based on morphological similarities, broad thorny ganglion cells have been proposed to be homologs of rabbit local edge detector ganglion cells, but we now show that the two cells have quite distinct physiological properties. Thus, our data argue against broad thorny cells as the homologs of local edge detector cells.

Detailed measurement of ganglion cell receptive fields often reveals significant deviations from a smooth, Gaussian profile. We studied the effect of these irregularities on the representation of fine spatial information in the retina. We recorded from nearby clusters of ganglion cells, testing their ability to determine the location of small flashed spots, and we compared the results to the prediction of a Gaussian receptive field model derived from reverse correlation. Despite considerable receptive field overlap, almost all ganglion cell pairs signaled nearly independently. For groups of five cells with highly overlapping receptive fields, the measured light-evoked currents encoded ∼33% more information than predicted by the Gaussian receptive field model. Including measured local irregularities in the receptive field model increased performance to the level observed experimentally. These results suggest that instead of being an unavoidable defect, irregularities may be a positive design feature of population neural codes.

In the primate visual system approximately 20 morphologically distinct pathways originate from retinal ganglion cells and project in parallel to the lateral geniculate nucleus (LGN) and/or the superior colliculus. Understanding of the properties of these pathways and the significance of such extreme early pathway diversity for later visual processing is limited. In a companion study we found that the magnocellular LGN-projecting parasol ganglion cells also projected to the superior colliculus and showed Y-cell receptive field structure supporting the hypothesis that the parasol cells are analogous to the well studied alpha-Y cell of the cat's retina. We here identify a novel ganglion cell class, the smooth monostratified cells, that share many properties with the parasol cells. Smooth cells were retrogradely stained from tracer injections made into either the LGN or superior colliculus and formed inner-ON and outer-OFF populations with narrowly monostratified dendritic trees that surprisingly appeared to perfectly costratify with the dendrites of parasol cells. Also like parasol cells, smooth cells summed input from L- and M-cones, lacked measurable S-cone input, showed high spike discharge rates, high contrast and temporal sensitivity, and a Y-cell type nonlinear spatial summation. Smooth cells were distinguished from parasol cells however by smaller cell body and axon diameters but ∼2 times larger dendritic tree and receptive field diameters that formed a regular but lower density mosaic organization. We suggest that the smooth and parasol populations may sample a common presynaptic circuitry but give rise to distinct, parallel achromatic spatial channels in the primate retinogeniculate pathway.

The distinctive parasol ganglion cell of the primate retina transmits a transient, spectrally nonopponent signal to the magnocellular layers of the lateral geniculate nucleus. Parasol cells show well-recognized parallels with the alpha-Y cell of other mammals, yet two key alpha-Y cell properties, a collateral projection to the superior colliculus and nonlinear spatial summation, have not been clearly established for parasol cells. Here, we show by retrograde photodynamic staining that parasol cells project to the superior colliculus. Photostained dendritic trees formed characteristic spatial mosaics and afforded unequivocal identification of the parasol cells among diverse collicular-projecting cell types. Loose-patch recordings were used to demonstrate for all parasol cells a distinct Y-cell receptive field "signature" marked by a nonlinear mechanism that responded to contrast-reversing gratings at twice the stimulus temporal frequency [second Fourier harmonic (F2)] independent of stimulus spatial phase. The F2 component showed high contrast gain and temporal sensitivity and appeared to originate from a region coextensive with that of the linear receptive field center. The F2 spatial frequency response peaked well beyond the resolution limit of the linear receptive field center, showing a Gaussian center radius of approximately 15 microm. Blocking inner retinal inhibition elevated the F2 response, suggesting that amacrine circuitry does not generate this nonlinearity. Our data are consistent with a pooled-subunit model of the parasol Y-cell receptive field in which summation from an array of transient, partially rectifying cone bipolar cells accounts for both linear and nonlinear components of the receptive field.

The development of retinal implants for the blind depends crucially on understanding how neurons in the retina respond to electrical stimulation. This study used multielectrode arrays to stimulate ganglion cells in the peripheral macaque retina, which is very similar to the human retina. Analysis was restricted to parasol cells, which form one of the major high-resolution visual pathways in primates. Individual cells were characterized using visual stimuli, and subsequently targeted for electrical stimulation using electrodes 9–15 μm in diameter. Results were accumulated across 16 ON and 9 OFF parasol cells. At threshold, all cells responded to biphasic electrical pulses 0.05–0.1 ms in duration by firing a single spike with latency lower than 0.35 ms. The average threshold charge density was 0.050 ± 0.005 mC/cm2, significantly below established safety limits for platinum electrodes. ON and OFF ganglion cells were stimulated with similar efficacy. Repetitive stimulation elicited spikes within a 0.1 ms time window, indicating that the high temporal precision necessary for spike-by-spike stimulation can be achieved in primate retina. Spatial analysis of observed thresholds suggests that electrical activation occurred near the axon hillock, and that dendrites contributed little. Finally, stimulation of a single parasol cell produced little or no activation of other cells in the ON and OFF parasol cell mosaics. The low-threshold, temporally precise, and spatially specific responses hold promise for the application of high-density arrays of small electrodes in epiretinal implants.

Synaptic circuits in primate fovea have been quantified for midget/parvocellular ganglion cells. Here, based on partial reconstructions from serial electron micrographs, we quantify synaptic circuits for two other types of ganglion cell: the familiar parasol/magnocellular cell and a smaller type, termed "garland." The excitatory circuits both derive from two types of OFF diffuse cone bipolar cell, DB3 and DB2, which collected unselectively from at least 6 +/- 1 cones, including the S type. Cone contacts to DB3 dendrites were usually located between neighboring triads, whereas half of the cone contacts to DB2 were triad associated. Ribbon outputs were as follows: DB3, 69 +/- 5; DB2, 48 +/- 4. A complete parasol cell (30 microm dendritic field diameter) would collect from approximately 50 cones via approximately 120 bipolar and approximately 85 amacrine contacts; a complete garland cell (25 microm dendritic field) would collect from approximately 40 cones via approximately 75 bipolar and approximately 145 amacrine contacts. The bipolar types contributed differently: the parasol cell received most contacts (60%) from DB3, whereas the garland cell received most contacts (67%) from DB2. We hypothesize that DB3 is a transient bipolar cell and that DB2 is sustained. This would be consistent with their relative inputs to the brisk-transient (parasol) ganglion cell. The garland cell, with its high proportion of DB2 inputs plus its high proportion of amacrine synapses (70%) and dense mosaic, might correspond to the local-edge cell in nonprimate retinas, which serves finer acuity at low temporal frequencies. The convergence of S cones onto both types could contribute S-cone input for cortical areas primary visual cortex and the middle temporal area.

An antibody against recoverin, the calcium-binding protein, labels photoreceptors, cone bipolar cells, and a subpopulation of cells in the ganglion cell layer. In the present study, we sought to establish the origin and identity of the cells expressing recoverin in the ganglion cell layer of the rat retina. By double labeling with rhodopsin, we demonstrate that early in development some of the recoverin-positive cells in the ganglion cell layer are photoreceptors. During the first postnatal week, these rhodopsin-positive cells are eliminated from the ganglion cell layer, but such neurons remain in the inner nuclear layer well into the first postnatal month. Another contingent of recoverin-positive cells, with morphological features equivalent to those of bipolar cells, is present in the postnatal retina, and approximately 50% of these neurons survive to maturity. The incidence of such cells in the ganglion cell layer was not affected by early transection of the optic nerve, a manipulation that causes rapid loss of retinal ganglion cells. These recoverin-positive cells were not double-labeled by cell-specific markers expressed by photoreceptors, rod bipolar cells, or horizontal and amacrine cells. Based on their staining with recoverin and salient morphological features, these ectopic profiles in the ganglion cell layer are most likely cone bipolar cells. Collectively, the results provide evidence for photoreceptors in the ganglion cell and inner nuclear layers of the developing retina, and a more permanent subpopulation of cone bipolar cells displaced to the ganglion cell layer.

We report a survey of the population of ganglion cells in the rabbit retina. A random sample of 301 neurons in the ganglion cell layer was targeted for photofilling, a method in which the arbors of the chosen neurons are revealed by diffusion of a photochemically induced fluorescent product from their somas. An additional 129 cells were labeled by microinjection of Lucifer yellow. One hundred and thirty-eight cells were visualized by expression of the gene encoding a green fluorescent protein, introduced by particle-mediated gene transfer. One hundred and sixty-six cells were labeled by particle-mediated introduction of DiI. In the total population of 734 neurons, we could identify 11 types of retinal ganglion cell. An analysis based on retinal coverage shows that this number of ganglion cell types would not exceed the available total number of ganglion cells. Although some uncertainties remain, this sample appears to account for the majority of the ganglion cells present in the rabbit retina. Some known physiological types could easily be mapped onto structural types, but half of them could not; a large set of poorly known codings of the visual input is transmitted to the brain.

A search for POU domain sequences expressed in the human retina has led to the identification of three closely related genes: Brn-3a, Brn-3b, and Brn-3c. The structure and expression pattern of Brn-3b was reported earlier (Xiang et al., 1993); we report here the structures and expression patterns of Brn-3a and Brn-3c. Antibodies specific for each Brn-3 protein were generated and shown to label only ganglion cells in a variety of vertebrate retinas. A complex pattern of strongly and weakly immunolabeled ganglion cells was observed in mouse, cat, and monkey retinae. In mouse and cat retinae, Brn-3a and Brn-3b proteins are found in a large fraction of ganglion cells, whereas Brn-3c is present in fewer ganglion cells. In the cat retina, anti-Brn-3a immunoreactivity was strong in the small ganglion cells (gamma cells) and weak in the remaining ganglion cells (alpha and beta cells); anti- Brn-3b immunoreactivity was present in all ganglion cells; and anti- Brn3c immunoreactivity was confined to the small ganglion cells. Immunolabeling of macaque retinae following retrograde labeling from the lateral geniculate nucleus revealed strong anti-Brn-3a immunoreactivity in a minority of retrogradely labeled P-type ganglion cells, and weak Brn-3a immunoreactivity in all of the remaining P- and M-type ganglion cells. In the same retinae, strong anti-Brn-3b immunoreactivity was seen in nearly all P-type ganglion cells and weak immunoreactivity in nearly all M-type ganglion cells. Each of the Brn-3- specific antibodies also labeled subsets of neurons in the dorsal root and trigeminal ganglia, suggesting that primary somatosensory neurons and retinal ganglion cells share genetic regulatory hierarchies. In vitro selection of an optimal DNA binding site using the Brn-3b POU domain has revealed a consensus [(A/G)CTCATTAA(T/C)] that is recognized by each of the Brn-3 POU domains and is distinct from binding sites previously described for other POU domain proteins.

We studied the relationship between the receptive fields of directionally selective retinal ganglion cells and the dendritic arbors of the same cells. The cells were recorded from extracellularly under visual control and then injected with Lucifer yellow. The arbor of Lucifer-filled dendrites could then be directly compared with the properties of the receptive field. A large population of on-off directionally selective cells was injected and drawn. The directionally selective ganglion cells had bistratified receptive fields similar to those previously described by others in the central retina. In the periphery, the dendritic fields became larger, rounder, and sparser than centrally. The diameters of the dendrites were measured in living or lightly fixed retinas; they were found to be somewhat larger than previously estimated by electron microscopy. The local structure of the dendritic arbor bore no obvious relation to the directional properties of the cell. The receptive fields of most cells were centered symmetrically around their dendritic fields. For about 10% of the cells, however, the receptive field was displaced. The displacement was always toward the preferred direction, relative to the dendritic field. The meaning of these shifts is not clear. In both cases, the diameter of the receptive field exceeded the diameter of the dendritic field only slightly; in our sample, the diameters of the receptive fields averaged 6% larger than the dendritic fields. This means that the neurons afferent to the directionally selective ganglion cells must either have narrow dendritic fields or, if they are wide spreading, have dendrites that do not conduct effectively along their length. It also means that the observed spread of neurobiotin between DS ganglion cells (Vaney, 1991) must be due to a very few gap junctions, or to some mechanism other than a gap junction.

We have examined in vitro the morphology and visual response properties of retinal ganglion cells innervating a component of the cat's lateral geniculate nucleus known as the geniculate wing (or retinorecipient zone of the pulvinar). Ganglion cells were first labeled in situ by retrograde transport of fluorescent microspheres from the geniculate wing. Labeled cells were injected intracellular with Lucifer yellow and biocytin in the isolated retina and visualized immunohistochemically. With one exception, stained cells appeared to belong to a single morphological class that corresponded closely to the epsilon cell of earlier descriptions (Leventhal et al., 1980; Rodieck and Watanabe, 1986). They had somas comparable in size to those of beta cells and large, sparse dendritic trees that ramified in the inner (ON) sublayer of the inner plexiform layer. Dendritic fields increased in size with eccentricity, but only within the central retina, and were among the largest so far reported for cat ganglion cells, exceeding those of alpha cells at most eccentricities. Dendritic profiles were typically elliptical with long axes pointing toward the area centralis. Axons were about as thick as those of beta cells and thicker than those of other varieties of non-alpha, non-beta ganglion cells. We recorded extracellularly from microsphere-labeled wing-projecting ganglion cells in a superfused, flattened eyecup preparation. All such cells exhibited sustained responses to standing contrast and had very large, concentric receptive fields with ON-centers and OFF-surrounds. Their response to gratings showed that they have relatively poor spatial resolution and a moderate amount of nonlinearity of spatial summation. These cells thus have many physiological response properties in common with ganglion cells previously termed “on-center tonic W-cells,” “on-center sluggish sustained cells,” and “Q-cells.” These findings indicate that ganglion cells innervating the cat's geniculate wing form a structurally and functionally homogeneous class. Their large dendritic and receptive fields and low-pass spatial frequency tuning suggest that fine spatial resolution is not required for the execution of their functional role(s).

Like the retinal ganglion cells from which they receive their input, most relay neurons in the lateral geniculate nucleus have ON- or OFF- center receptive fields with antagonistic surrounds. In the cat, neurons with these two types of receptive fields are anatomically intermingled, even though the ON and OFF systems are functionally segregated. In the ferret, there is a sublamination of the retinal input to lateral geniculate nucleus laminae A and A1. We have investigated the function of this sublamination by making microelectrode recordings and have found that each sublamina consists of geniculate neurons of a single center type.

Bilaterally projecting retinal ganglion cells (BPRGCs) in the adult Syrian golden hamster were identified through the use of two retrogradely transported neuronal labels, horseradish peroxidase and Nuclear Yellow, placed separately in each optic tract. The distribution and size of doubly labeled retinal ganglion cells were characterized and their numbers were determined. Strict criteria were used to exclude artifactual doubly labeled cells. This work revealed that: (a) BPRGCs comprise less than 0.01% of the entire retinal ganglion cell population, averaging 7.4 (SD = 3) cells per retina; (b) BPRGCs are found primarily in the upper, peripheral retina and not along the vertical meridian or in the temporal crescent; and (c) BPRGCs correspond in size to ordinary retinal ganglion cells in their immediate vicinity, thus providing no evidence that they comprise a separate population of cells. Electrophysiological collision experiments were also performed, with stimulating electrodes in the two brachia of the superior colliculi and a recording electrode in one optic nerve. A collision effect was not detected, thus supporting the anatomical findings of rare bilateral branching of optic nerve axons. The occurrence of BPRGCs may reflect occasional ambiguities in the cues that guide axons through the chiasm.

The degree to which a retinal ganglion cell's environment can affect its morphological development was studied by manipulating the distribution of ganglion cells in the developing cat retina. In the newborn kitten there is an exuberant ganglion cell projection from temporal retina to the contralateral lateral geniculate nucleus (LGNd) (Leventhal et al., 1988) and from nasal retina to the ipsilateral LGNd. Neonatal, unilateral optic tract section results in the survival of many of these ganglion cells (Leventhal et al., 1988). The morphology of ganglion cells which survive in regions of massively reduced ganglion cell density was studied. As reported previously (Linden and Perry, 1982; Perry and Linden, 1982; Ault et al., 1985; Eysel et al., 1985), we found that the dendritic fields of all types of ganglion cells on the border of an area depleted of ganglion cells extended into the depleted area. The cell bodies and dendritic fields of alpha and beta cells within depopulated areas, as well as on the borders of the depopulated areas, were larger than normal. The dendritic fields of these cells also exhibited abnormal branching patterns. For alpha and beta cell types the relative increase in size tended to be greatest where the relative change in density was the greatest. In fact, isolated beta cells within the cell-poor area centralis region resembled normal central alpha cells in the cell-rich region of the area centralis in the same retina. Interestingly, in the same regions of reduced density where alpha and beta cells were dramatically larger than normal, the cell body and dendritic field sizes of other cell types (epsilon, g1 and g2 were unchanged. These results indicate that neuronal interactions during development contribute to the morphological differentiation of retinal ganglion cells and that different mechanisms mediate the morphological development of different classes of cells in cat retina.

We have studied the development of retinal ganglion cell morphology in the cat's visual system from early fetal to postnatal times. In particular, we have examined the contribution of growth and remodeling to the establishment of mature retinal ganglion cell form. Ganglion cells were identified by retrograde labeling with rhodamine latex microspheres deposited in the superior colliculus and lateral geniculate nucleus between embryonic day 34 (E34; birth = E65) and adulthood. To reveal the fine morphological details of retrogradely labeled ganglion cells, 48 hr later Lucifer yellow was injected intracellularly in living retinae that had been dissected and maintained in vitro. Our results show that at E35–37 the majority of ganglion cells are very simple in morphology, with a few dendritic processes that are generally aligned in a radial direction towards or away from the optic disc. During the ensuing 2 week period, there is a progressive growth and elaboration of dendrites. By E50, some ganglion cells resembling the adult alpha, beta, and gamma classes can be identified based on comparisons of the appearance and dimensions of their dendritic trees and somata with neighboring filled cells. However, ganglion cell dendrites and axons at this age express several transient morphological features. The axons of ganglion cells give rise to delicate processes originating from the intraretinal portion of the axon, including side branches, present in about half of the cells, and occasionally bifurcations that give rise to axon collaterals. These transient axonal features are present throughout development, including the neonatal period; no axon collaterals were observed after postnatal day 15, while axonal side branches persisted even at P31 but were gone by adulthood. Ganglion cell dendrites exhibit excessive branches and exuberant somatic and dendritic spines. Quantitative analysis of these processes shows that after E45 dendritic trees increase dramatically in complexity, reaching the peak number of spines and branch points by the first week of postnatal life. The number of dendritic processes then falls abruptly to reach near-adult levels by the end of the first postnatal month. Even though dendritic morphology closely resembles that seen in the adult at this age, ganglion cell bodies and dendrites must continue to grow to reach their adult size.(ABSTRACT TRUNCATED AT 400 WORDS)

The ganglion cells of the cat's retina form several classes distinguishable in terms of soma size, axon diameter, dendritic morphology, physiological properties, and central connections. Labeling with [3H]thymidine shows that the ganglion cells which survive in the adult are produced as several temporally shifted, overlapping waves: medium-sized cells are produced before large cells, whereas the smallest ganglion cells are produced throughout the period of ganglion cell generation (Walsh, C., E. H. Polley, T. L. Hickey, and R. W. Guillery (1983) Nature 302: 611–614). Large cells and medium-sized cells show the same distinctive pattern of production, forming rough spirals around the area centralis. The oldest cells tend to lie superior and nasal to the area centralis, whereas cells in the inferior nasal retina and inferior temporal retina are, in general, progressively younger. Within each retinal quadrant, cells nearer the area centralis tend to be older than cells in the periphery, but there is substantial overlap. The retinal raphe divides the superior temporal quadrant into two zones with different patterns of cell addition. Superior temporal retina near the vertical meridian adds cells only slightly later than superior nasal retina, whereas superior temporal retina near the horizontal meridian adds cells very late, contemporaneously with inferior temporal retina. The broader wave of production of smaller ganglion cells seems to follow this same spiral pattern at its beginning and end. The presence of the area centralis as a nodal point about which ganglion cell production in the retinal quadrants pivots suggests that the area centralis is already an important retinal landmark even at the earliest stages of retinal development. This sequence of ganglion cell production differs markedly from that seen in the retinae of nonmammalian vertebrates, where new ganglion cells are added as concentric rings to the retinal periphery, and also bears no simple relationship to the cat's retinal decussation line. However, it can be related in a straightforward manner to the organization of axons in the cat's optic tract, suggesting that the fiber order in the tract represents a grouping of fibers by age.