Morphological types of horizontal cell in the retina of the domestic cat.

Two morphologically distinct types of horizontal cell are described from Golgi-stained whole mounts of the cat retina. They are referred to as A-type and B-type cells. The two types differ in their dendritic branching pattern, their overall size and the absence or presence of an axon. At every retinal position the dendrites of B-type cells branch more densely and overlap each other more frequently than do the dendrites of A-type cells. At equivalent retinal positions the dendritic field size of A-type cells is greater than that of B-type cells by a factor of about 1.5. Only B-type cells have an axon, which branches at the end into a large axon terminal system. The axons have no preferred direction of orientation. The stain-ability of horizontal cells by different Golgi methods is discussed.     

The morphology and topographic distribution of AII amacrine cells in the cat retina

When cat retina is incubated in vitro with the fluorescent dye, 4',6-diamidino-2-phenyl-indole (DAPI), a uniform population of neurons is brightly labelled at the inner border of the inner nuclear layer. The dendritic morphology of the DAPI-labelled cells was defined by iontophoretic injection of Lucifer yellow under direct microscopic control: all the filled cells had the narrow-field bistratified morphology that is distinctive of the AII amacrine cells previously described from Golgi-stained retinae. Although the AII amacrines are principal interneurons in the rod-signal pathway, their density distribution does not follow the topography of the rod receptors, but peaks in the central area like the cone receptors and the ganglion cells. There are some 512 000 AII amacrines in the cat retina and their density ranges from 500 cells per square millimetre at the superior margin to 5300 cells per square millimetre in the centre (retinal area is 450 mm2). The isodensity contours are kite-shaped, particularly at intermediate densities, with a horizontal elongation towards nasal retina. The cell body size and the dendritic dimensions of AII amacrines increase with decreasing cell density. The lobular dendrites in sublamina a of the inner plexiform layer span a restricted field of 16-45 microns diameter, while the arboreal dendrites in sublamina b form a varicose tree of 18-95 microns diameter. The dendritic field coverage of the lobular appendages is close to 1.0 (+/- 0.2) at all eccentricities whereas the coverage of the arboreal dendrites doubles within the first 1.5 mm and then remains constant at 3.8 (+/- 0.7) throughout the periphery.     

Morphology and topography of on- and off-alpha cells in the cat retina

Neurofibrillar staining methods were found to stain all alpha cells of the cat retina completely, that is the perikaryon, the axon and the dendritic branches. The dendrites of the alpha cells in vertical sections were found to be unistratified and to occupy two narrow strata in the outer half of the inner plexiform layer. This difference in branching level could also be observed in whole-mount preparations and it has been demonstrated in the preceding paper (Peichl & W?ssle 1981) that it corresponds to the physiological on-off dichotomy. Thus the topographical distribution of on- and off-alpha cells could be studied. They are found to occur in about equal numbers. Both on- and off-alpha cell perikarya form a regular lattice and both lattices are superimposed independently. The dendritic branches of neighbouring alpha cells overlap and each retinal point is covered by the dendritic field of at least one on- and one off-alpha cell. The dendritic trees of on-alpha cells seem to have more small branches and are on the average smaller than those of off-alpha cells. The density of alpha cells was found to peak in the central area whence it continuously decreased towards the retinal periphery.     

Receptor contacts of horizontal cells in the retina of the domestic cat.

The terminal aggregations of A- and B-type horizontal cells, stained by the Golgi-Colonnier method, have been analysed. The pattern of the aggregations is regular and is shown to be in register with the cone mosaic. Both tyes of horizontal cell are in contact with at least 80% of the cones above their dendritic fields. Therefore, the different horizontal cell classes cannot be selective for a special kind of cone but must have at least 60% of the cone input in common. Each A-type horizontal cell makes contacts with between 120 and 170 cones, and each B-type horizontal cell with 60-90 cones. An individual A-type horizontal cell occupies an average of 20% of the lateral elements of the triads in a cone pedicle, but an individual B-type cell fills only some 13%. Each and every cone is connected with several of both types of horizontal cell. An estimation of the number of rods converging onto a single axon terminal system showed that it could be as many as 3000.     

The formation of the area centralis of the retinal ganglion cell layer in the chick

In adult domestic chickens, the neurones in the retinal ganglion cell layer are very unevenly disposed such that there is a sixfold increase in neurone density from the retinal edge to the retinal centre. The formation of the high ganglion-cell-density area centralis was studied on chick retinal wholemounts from the 8th day of incubation (E8) to 4 weeks after hatching (4WAH). The density of viable neurones and the number and the distribution of pyknotic neurones in the ganglion cell layer were estimated across the whole retina. Between E8 and E10, the distribution of neurones in the ganglion cell layer was anisodensitic with 53,000 mm-2 in the centre compared to 34,000 mm-2 in the periphery of the retina. Thereafter, a progressively steeper gradient of neurone density developed, which decreased from 24,000 mm-2 in the retinal centre to 6000 mm-2 at the retinal periphery by 4WAH. Neuronal pyknosis in the ganglion cell layer was observed between E9 and E17. From E11 onwards, consistently more pyknotic neurones were found in the peripheral than in the central retina. It was estimated that over the period of cell death approximately twice as many neurones died per unit area in the retinal periphery than in the centre. Retinal area measurements and estimation of neurone densities in the ganglion cell layer after the period of neurone generation and neurone death indicated differential retinal expansion, with more expansion in the peripheral than in the central retina. These observations allow us to conclude that the formation of the area centralis of the chick retina involves (1) slightly higher cell generation in the retinal centre, (2) higher rate of cell loss in the retinal periphery and (3) differential retinal expansion.     

Horizontal cells of the rabbit retina: some quantitative properties revealed by selective staining

Horizontal cells of the rabbit retina were selectively stained by demonstration of beta-hydroxybutyrate dehydrogenase activity. Thereafter, the size of the cell bodies, the distance between neighbouring cells, and the number of cells per mm2 were measured. In the area centralis, the horizontal cell bodies occupy only 4.2% of the total retinal area; in the far periphery, however, 12.8% of the retinal area consist of horizontal cell bodies. Furthermore, the horizontal cells of the retinal periphery have much larger cell bodies as compared with those of the retinal center. The far periphery, for these reasons, is concluded to be the optimal region for intracellular microelectrode recordings from rabbit retinal horizontal cells.     

The shape and arrangement of the cholinergic neurons in the rabbit retina

The acetylcholine-synthesizing neurons of the rabbit retina were selectively stained by intraocular injection of the fluorescent dye 4,6-diamidino-2-phenylindole (DAPI). Retinas were then isolated from the eye, fixed for 10-30 min with 4% paraformaldehyde, and mounted flat on the stage of a fluorescence microscope. The acetylcholine-synthesizing cells were penetrated under visual control by microelectrodes filled with lucifer yellow CH. When the dye was electrophoretically injected into the cells, complete filling of their dendrites often occurred. Cells were successfully injected as long as one month after fixation of the tissue. Complete or nearly complete filling of 281 cells was accomplished, at retinal locations systematically covering the retinal surface. The cells stained with DAPI were found to form a single morphological population. They have two to seven primary dendrites, which branch repeatedly within a narrow plane and form a round or slightly oval dendritic tree. The branching becomes very fine for the distal one third of the dendritic tree, and the dendrites there are studded with small swellings. The distal dendritic tree lies mainly within one of the two thin strata of the inner plexiform layer where acetylcholine is present. The shape and size of the dendritic tree are continuously graded across the retina, the dendritic tree is narrower and the branching denser in the central retina, wider and sparser in the periphery. From knowledge of the population density and the shape of the neurons, one can reconstruct the array of dendrites that exists within the inner plexiform layer. The overlap of the dendritic fields is an order of magnitude greater than of any other retinal neuron previously described. Because the cells not only overlap widely but branch quite profusely, a very dense plexus of cholinergic dendrites is created.     

Functional anatomy of the photoreceptor and second-order cell mosaics in the retina of Xenopus laevis

Mosaics of photoreceptors, and horizontal and bipolar cells of the Xenopus laevis retina were studied in whole-mount preparations applying lectin-cytochemical, immunocytochemical and intracellular labeling techniques. The combined density of all photoreceptor types was about 13700/mm2, of which rods represented 53%. Of the cones, the large long-wavelength-sensitive (86% of all cones) and the miniature ultraviolet-wavelength-sensitive (4%) ones could be labeled with peanut agglutinin, whereas the large short-wavelength-sensitive (10%) cones remained unlabeled. There were no significant regional differences in photoreceptor distribution. Bipolar cells were selectively labeled with antibodies against calretinin. Their density was between 4000 and 6000 cells/cm2, with slightly elevated numbers in the superior nasal quadrant. Two types of horizontal cell were injected intracellularly. The luminosity-type cells were more frequent (approximately 1000 cells/mm2) than the chromaticity cells (approximately 450 cells/mm2). The dendritic field size of the latter cell type was threefold bigger than that of the luminosity cells. The coverage factors were estimated to be 3.3 for the luminosity cells and 5.2 for the chromaticity cells. The luminosity cells contacted all photoreceptor types, whereas chromatic horizontal cells received their inputs from the short-wavelength-sensitive cones and from some, but not all, rods. Luminosity cells encounter about 50-60 potential synaptic partners within their dendritic fields, whereas chromatic horizontal cells only about 20. Chromatic horizontal cells form multiple synaptic contacts with the short-wavelength-sensitive cones. The results indicate that the overall photoreceptor to bipolar and bipolar to ganglion cell convergence in Xenopus retina is similar to that in the central retinal specialized regions of mammals, predicting comparable spatial resolutions.     

Cadherin is required for dendritic morphogenesis and synaptic terminal organization of retinal horizontal cells

Dendrite morphology of neurons provides a structural basis for their physiological characteristics, and is precisely regulated in a cell type-dependent manner. Using a unique transposon-mediated gene transfer system that enables conditional and cell-type specific expression of exogenous genes, we investigated the role of cadherin on dendritic morphogenesis of horizontal cells in the developing chicken retina. We first visualized single horizontal cells by overexpressing membrane-targeted EGFP, and confirmed that there were three subtypes of horizontal cells, the dendritic terminals of which projected to distinct synaptic sites in the outer plexiform layer. Expression of a dominant-negative cadherin decreased the dendritic field size, and perturbed the termination of dendritic processes onto the photoreceptor cells. The cadherin blockade also impaired the accumulation of GluR4, a postsynaptic marker, at the cone pedicles. We thus provide in vivo evidence that cadherin is required for dendrite morphogenesis of horizontal cells and subsequent synapse formation with photoreceptor cells in the vertebrate retina.     

Nuclear lamins are differentially expressed in retinal neurons of the adult rat retina

Lamins are type V intermediate filament proteins that support nuclear membranes. They are divided into A-type lamins, which include lamin A and C, and B-type lamins, which include lamin B1 and B2. In the rat brain, lamin A and C are expressed in relatively equal amounts, while the expressions of lamin B1 and B2 vary depending on the cell type. Lamins play important roles in normal morphogenesis and function. In the nervous system, their abnormal expression causes several neurodegenerative diseases such as peripheral neuropathy, leukodystrophy and lissencephaly. The retina belongs to the central nervous system (CNS) and has widely been used as a source of CNS neurons. We investigated the expression patterns of lamin subtypes in the adult rat retina by immunohistochemistry and found that the staining patterns differed when compared with the brain. All retinal neurons expressed lamin B1 and B2 in relatively equal amounts. In addition, horizontal cells and a subpopulation of retinal ganglion cells expressed lamin A and C, while photoreceptor cells expressed neither lamin A nor C, and all other retinal neurons expressed lamin C only. This differential expression pattern of lamins in retinal neurons suggests that they may be involved in cellular differentiation and expression of cell-specific genes in individual retinal neurons.     

The structure and diversity of retinal ganglion cells in the masked greenling Hexagrammos octogrammus Pallas, 1814 (Pisces: Scorpaeniformes: Hexagrammidae)

The authors studied the structure and diversity of retinal ganglion cells (GC) in the masked greenling Hexagrammos octogrammus. In vivo labelling with horseradish peroxidase revealed GCs of various structures in retinal wholemounts. A total of 154 cells were camera lucida drawn, and their digital models were generated. Each cell was characterized by 17 structural and topological parameters. Using nine clustering algorithms, a variety of clusterings were obtained. The optimum clustering was found using silhouette analysis. It was based on a set of three variables associated with dendritic field size and dendrite stratification depth in the retina. A total of nine cell types were discovered. A number of non-parametric tests showed significant pair-wise between-cluster differences in at least four parameters with medium and large effect sizes. Three large-field types differed mainly in dendritic field size, total dendrite length, level of dendrite stratification in the retina and position of somata. Six medium- to small-field types differed mainly in the structural complexity of dendritic arbors and level of dendrite arborization. Cells similar and obviously homologous to types 1–4 were identified in many fish species, including teleosts. Potential homologues of type 5 cells were identified in fewer teleost species. Cells similar to types 6–9 in relative dendritic field size and dendrite arborization pattern were also described in several teleostean species. Nonetheless, their homology is more questionable as their stratification patterns do not match so well as they do in large types. Potential functional matches of the GC types were identified in a number of teleostean species. Type 1 and 2 cells probably match spontaneously active units with the large receptive field centre, so-called dimming and lightening detectors; type 4 may be a counterpart of changing contrast detectors with medium receptive field centre size preferring fast-moving stimuli. Type 3 (biplexiform) cells have no obvious functional matches. Probable functional matches of types 6, 8 and 9 belong to ON-centre elements with small receptive fields such as ON-type direction-selective cells, ON-type spot detectors or ON-type spontaneously active units. Type 5 and 7 cells may match ON–OFF type units, in particular, changing contrast detectors or orientation-selective units. Potential functional matches of GC types presently described are involved in a wide spectrum of visual reactions related to adaptation to gradual change in illumination, predator escape, prey detection and capture, habitat selection and social behaviour.     

Organization of the outer plexiform layer of the primate retina: electron microscopy of Golgi-impregnated cells

Golgi-impregnated retinae of rhesus monkeys have been examined by serial section electron microscopy to establish in a quantitative manner the neural connexions in the outer plexiform layer. The results have shown that there are two types of midget bipolar cell, here called the invaginating midget bipolar and the flat midget bipolar. Both types of midget bipolar are exclusive to a single cone. The invaginating midget bipolar has been found to fit a dendritic terminal process into every invagination in the cone pedicle base. The flat midget bipolar has dendritic terminals that make superficial contact on the cone pedicle base. There are twice as many dendritic terminals and points of contact with the cone pedicle on a flat midget bipolar top as compared with an invaginating midget bipolar top. These observations, together with light microscope counts of the numbers of the two types of midget bipolars, suggest that there are two midget bipolars per cone. The diffuse cone bipolar (the flat bipolar) also makes superficial contacts on the cone pedicle base, and serial sections have shown that a flat bipolar contacts about six cones. Rod bipolars connect exclusively to rods and their dendritic terminals always end as one of the central processes that penetrate the invagination. Horizontal cell dendrites end exclusively in cone pedicles and their axon terminals end in rod spherules. The point of contact with both the types of receptor is as the lateral elements of the invaginations. A single small horizontal cell contacts about seven cones and a large horizontal cell contacts about twelve cones. The numbers of contacts per cone pedicle decrease from the centre to the periphery of the horizontal cell's dendritic field, suggesting there is an overlap of four to six horizontal cells onto a single cone pedicle. The horizontal cell axon terminals are too numerous to assess in absolute numbers but there is only one terminal to a given rod spherule from any particular axon.     

Spatial Distribution of Excitatory Synapses on the Dendrites of Ganglion Cells in the Mouse Retina

Excitatory glutamatergic inputs from bipolar cells affect the physiological properties of ganglion cells in the mammalian retina. The spatial distribution of these excitatory synapses on the dendrites of retinal ganglion cells thus may shape their distinct functions. To visualize the spatial pattern of excitatory glutamatergic input into the ganglion cells in the mouse retina, particle-mediated gene transfer of plasmids expressing postsynaptic density 95-green fluorescent fusion protein (PSD95-GFP) was used to label the excitatory synapses. Despite wide variation in the size and morphology of the retinal ganglion cells, the expression of PSD95 puncta was found to follow two general rules. Firstly, the PSD95 puncta are regularly spaced, at 1–2 µm intervals, along the dendrites, whereby the presence of an excitatory synapse creates an exclusion zone that rules out the presence of other glutamatergic synaptic inputs. Secondly, the spatial distribution of PSD95 puncta on the dendrites of diverse retinal ganglion cells are similar in that the number of excitatory synapses appears to be less on primary dendrites and to increase to a plateau on higher branch order dendrites. These observations suggest that synaptogenesis is spatially regulated along the dendritic segments and that the number of synaptic contacts is relatively constant beyond the primary dendrites. Interestingly, we also found that the linear puncta density is slightly higher in large cells than in small cells. This may suggest that retinal ganglion cells with a large dendritic field tend to show an increased connectivity of excitatory synapses that makes up for their reduced dendrite density. Mapping the spatial distribution pattern of the excitatory synapses on retinal ganglion cells thus provides explicit structural information that is essential for our understanding of how excitatory glutamatergic inputs shape neuronal responses.     

Distribution pattern of three neural calcium-binding proteins (NCS-1, VILIP and recoverin) in chicken, bovine and rat retina

Summary Neural Ca²⁺-binding proteins (NCaPs) constitute a subfamily of 4-EF-hand proteins, and display a histological and structural dichotomy: the A-type NCaPs are selectively expressed by the retina and pineal organ and display two canonical EF-hands, whereas the B-type NCaPs are found in the entire brain and present three regular EF-hands. In this study, antisera were raised against the A-type NCaP recoverin (26 kDa) and the B-type NCaPs VILIP and NCS-1 (22 kDa). Since the sequence identity among NCaPs is high, specific polyclonal antibodies were purified by double cross-immunoaffinity chromatography; both ELISA and immunoblot analyses determined that the resulting antibodies showed selectivity ratios inferior to 1/363 for the two other related NCaPs. Besides, the anti-VILIP antibodies displayed some affinity toward neurocalcin δ, and the antirecoverin antibodies recognized a 24 kDa protein, which is most likely visinin. Thus, immunohistochemical studies on the chicken, rat and cow retina revealed that anti-recoverin antibodies recognized the vertebrate photoreceptors and a small number of mammalian bipolar cells. Anti-VILIP antibodies exclusively labelled the inner Retina, I.e. the amacrine and ganglion cells. NCS-1 was mainly present in the photoreceptor inner segments, the inner plexiform layer and the ganglion cells. NCS-1 showed the highest species disparity. The retinal localization of NCS-1 and VILIP offered an important morphological basis for the understanding of their function. Furthermore, specific antibodies against the NCaPs may enable the identification of cell populations in more complex neural tissues, such as the brain.     

Identification of a Circadian Clock-Controlled Neural Pathway in the Rabbit Retina

Background

Although the circadian clock in the mammalian retina regulates many physiological processes in the retina, it is not known whether and how the clock controls the neuronal pathways involved in visual processing.

Methodology/Principal Findings

By recording the light responses of rabbit axonless (A-type) horizontal cells under dark-adapted conditions in both the day and night, we found that rod input to these cells was substantially increased at night under control conditions and following selective blockade of dopamine D₂, but not D₁, receptors during the day, so that the horizontal cells responded to very dim light at night but not in the day. Using neurobiotin tracer labeling, we also found that the extent of tracer coupling between rabbit rods and cones was more extensive during the night, compared to the day, and more extensive in the day following D₂ receptor blockade. Because A-type horizontal cells make synaptic contact exclusively with cones, these observations indicate that the circadian clock in the mammalian retina substantially increases rod input to A-type horizontal cells at night by enhancing rod-cone coupling. Moreover, the clock-induced increase in D₂ receptor activation during the day decreases rod-cone coupling so that rod input to A-type horizontal cells is minimal.

Conclusions/Significance

Considered together, these results identify the rod-cone gap junction as a key site in mammals through which the retinal clock, using dopamine activation of D₂ receptors, controls signal flow in the day and night from rods into the cone system.     

A-type and B-type lamins initiate layer assembly at distinct areas of the nuclear envelope in living cells

To investigate nuclear lamina re-assembly in vivo, Drosophila A-type and B-type lamins were artificially expressed in Drosophila lamin Dm₀null mutant brain cells. Both exogenous lamin C (A-type) and Dm₀ (B-type) formed sub-layers at the nuclear periphery, and efficiently reverted the abnormal clustering of the NPC. Lamin C initially appeared where NPCs were clustered, and subsequently extended along the nuclear periphery accompanied by the recovery of the regular distribution of NPCs. In contrast, lamin Dm₀ did not show association with the clustered NPCs during lamina formation and NPC spacing recovered only after completion of a closed lamin Dm₀ layer. Further, when lamin Dm₀ and C were both expressed, they did not co-polymerize, initiating layer formation in separate regions. Thus, A and B-type lamins reveal differing properties during lamina assembly, with A-type having the primary role in organizing NPC distribution. This previously unknown complexity in the assembly of the nuclear lamina could be the basis for intricate nuclear envelope functions.     

Horizontal slice preparation of the retina

Traditionally the vertical slice and the whole-mount preparation of the retina have been used to study the function of retinal circuits. However, many of retinal neurons, such as amacrine cells, expand their dendrites horizontally, so that the morphology of the cells is supposed to be severely damaged in the vertical slices. In the whole-mount preparation, especially for patch-clamp recordings, retinal neurons in the middle layer are not easily accessible due to the extensive coverage of glial cell (Mueller cell) s endfeets. Here, we describe the novel slicing method to preserve the dendritic morphology of retinal neurons intact. The slice was made horizontally at the inner layer of the retina using a vibratome slicer after the retina was embedded in the low-temperature melting agarose gel. In this horizontal slice preparation of the retina, we studied the function of retinal neurons compared with their morphology, by using patch-clamp recording, calcium imaging technique, immunocytochemistry, and single-cell RT-PCR.     

Morphological alterations in retinal neurons in the S334ter-line3 transgenic rat

The S334ter-line-3 rat is a transgenic model of retinal degeneration developed to express a rhodopsin mutation similar to that found in human retinitis pigmentosa (RP) patients. Previous studies have focused on physiological changes in retinal cells and higher centers of the visual system with this model of retinal degeneration. However, little is known about the morphological changes in retinal cells during the development of the S334ter-line-3 rat. In order to understand and aid vision-rescue strategies, our aim has been to describe the retinal degeneration pattern in this model. We focus on changes in the morphologies of horizontal, bipolar, and amacrine cells in developing S334ter-line-3 rat retinas. Degeneration of photoreceptors begins in the central retina and progresses toward the periphery. In retinas at post-natal day 15 (P15), horizontal and rod bipolar cells show normal morphology. However, at P21, horizontal and rod bipolar cells exhibit abnormal processes at the outer plexiform layer, whereas the outer nuclear layer is significantly thinner. A glial reaction occurs concomitantly. In contrast, modifications in cone-bipolar and amacrine cells are much slower and do not occur until P90 and P180, respectively. The density of horizontal and rod-bipolar cells significantly drops after P60. Overall, the S334ter-line-3 model exhibits the hallmarks of cellular remodeling caused by photoreceptor degeneration. Its moderately fast time course makes the S334ter-line-3 a good model for studying vision-rescue strategies.     

Differential cell death and Bcl-2 expression in the mouse retina after glutathione decrease by systemic D,L-buthionine sulphoximine administration

Glutathione (GSH) plays a critical role in cellular defense against unregulated oxidative stress in mammalian cells including neurons. We previously demonstrated that GSH decrease using [D, L]-buthionine sulphoximine (BSO) induces retinal cell death, but the underlying mechanisms of this are still unclear. Here, we demonstrated that retinal GSH level is closely related to retinal cell death as well as expression of an anti-apoptotic molecule, Bcl-2, in the retina. We induced differential expression of retinal GSH by single and multiple administrations of BSO, and examined retinal GSH levels and retinal cell death in vivo. Single BSO administration showed a transient decrease in the retinal GSH level, whereas multiple BSO administration showed a persistent decrease in the retinal GSH level. Retinal cell death also showed similar patterns: transient increases of retinal cell death were observed after single BSO administration, whereas persistent increases of retinal cell death were observed after multiple BSO administration. Changes in the retinal GSH level affected Bcl-2 expression in the retina. Immunoblot and immunohistochemical analyses showed that single and multiple administration of BSO induced differential expressions of Bcl-2 in the retina. Taken together, the results of our study suggest that the retinal GSH is important for the survival of retinal cells, and retinal GSH appears to be deeply related to Bcl-2 expression in the retina. Thus, alteration of Bcl-2 expression may provide a therapeutic tool for retinal degenerative diseases caused by retinal oxidative stress such as glaucoma or retinopathy.     

Invariants of spatial summation for S (short wavelength) cone vision

Spatial summation in the human visual system was studied as a function of retinal eccentricity upon selective stimulation of the short-wavelength sensitive cones. The area of complete spatial summation (Ricco's area) was found to increase with retinal eccentricity while the threshold of stimuli equal in size with Ricco's area remained constant. Comparisons with known morphology of the small bistratified retinal ganglion cells, the only cells known to be excited by S-one ON stimulation, showed that Ricco's area included 2-4 such cells and is up to 1.5 times larger than the dendritic field of a single cell. These relationships were relatively constant within the eccentricity range tested (5-20 deg along the temporal horizontal meridian) and might be the source of threshold invariance of stimuli matching Ricco's area.