Cat Retina Research Articles

Local oxygen diffusion coefficients in the cat retina and cornea

Local oxygen diffusion coefficients in the cat retina and cornea

Labeling and distribution of AII amacrine cells in the rabbit retina

The fluorescent dye 4,6-diamino-2-phenylindole (DAPI) has previously been used to label starburst amacrine cells selectively in the rabbit retina and AII amacrine cells in the cat retina. Using the rabbit retina, we show that intraocular injection of DAPI labels starburst amacrine cells as seen 1-2 days later. In contrast, after a brief in vitro incubation with DAPI, AII amacrine cells are selectively labeled. Amacrine cells were identified by intracellular staining with Lucifer Yellow. AII amacrine cells are arranged in a regular mosaic with a density of 2,800 cells/mm2 near the visual streak declining to about 500 cells/mm2 in the far periphery. The coverage of the lobular dendritic field in sublamina alpha is approximately 1.5 across the retina, but the coverage of the fine dendritic field in sublamina b increases from 3 centrally to 4 in the inferior periphery, and to above 8 in the superior periphery.

Microcircuitry related to the receptive field center of the on-beta ganglion cell

1. We have investigated the anatomic basis for the Gaussian-like receptive field center of the on-beta ("X") ganglion cell in the area centralis of cat retina. Three adjacent on-beta cells were reconstructed from electron micrographs of 279 serial sections cut vertically through a patch of retina at approximately 1 degree eccentricity. 2. All the bipolar synapses on these cells were identified, and about one-half of these were traced to type b1 bipolar cells, which formed a regular array in the plane of the retina. 3. On average, seven b1 cells contributed to a beta cell: bipolar axons near the middle of the beta dendritic field tended to give many contacts (12-33 contacts); axons near the edge of the field tended to give few contacts (3-4 contacts). 4. Each b1 cell collected from four to seven cones, and the mean number of cones converging through the b1 array to a beta cell was 30. 5. Assuming equal effectiveness for all b1----beta cell synapses, a spatial weighting function was derived from these results. The mean radius of this function at 1/e amplitude for three beta cells was 18.0 +/- 1.1 (SD) microns. This is considerably narrower than the corresponding measurements of the beta cell receptive field center (28 +/- 3 microns) at this eccentricity. 6. It is concluded, in agreement with previous work, that all cones encompassed by the beta cell's dendritic field and those slightly beyond it connect directly to the beta cell via the b1 bipolar cell array. However, the center of the beta cell receptive field is still broader by approximately 60%. This suggests that pooling of cone signals may occur at the level of the outer plexiform layer.

Anatomical correlations between soma size, axon diameter, and intraretinal length for the alpha ganglion cells of the cat retina

Retinal ganglion cells within the same region of the retina may have different lengths of axon before reaching the optic disc depending on the route they take with respect to the temporal raphe. We have investigated whether there is a correlation between soma and intraretinal axon diameter and how these parameters relate to intraretinal axon length on both sides of the cat temporal raphe. Retinas were wholemounted and alpha-cell somata and fibers stained with a modified neurofibrillar method. Moving peripherally from the area centralis along the raphe there was a progressively increasing difference between the intraretinal axon lengths for nearly adjacent cells across the raphe, which reached a maximum of 4-5 mm at the retinal periphery. Cells on the nasal aspect of the raphe had shorter axons than did adjacent cells on the temporal aspect of the raphe. Comparison of soma diameter samples across the raphe showed there was no clear trend between soma diameter and intraretinal length. Replotting the raphe and sample areas on a cell density map indicated that differences in soma diameter could be attributed to ganglion-cell density differences between the sampled areas. Examination of the stained cells revealed that within the initial length of the axon there was a region showing a reduction of axon diameter (diameter less than 1 micron), which varied in length from cell to cell. The axon was, therefore, divided into three segments: the portion of axon prior to thinning (A), the thin segment itself (B), and the part of the axon after the thin segment (C). The diameter of each segment (A,B,C) and the lengths of the first and second segments (A,B) were significantly correlated with soma diameter (P less than 0.001). From measurements of the axon diameter of segment C, it was concluded that alpha-cell axons continue to increase in diameter along their path towards the optic disc. The present report indicates that alpha-cell soma size, when going from the area centralis to the periphery along the raphe, reaches a plateau and then declines within more peripheral retinal locations in spite of increasing intraretinal axon length. Thus, there is no positive correlation between soma or axon diameter and intraretinal axon length. The anatomical findings are discussed in relation to previous reports of retinal development and complementary conduction times within intraretinal and extraretinal visual pathways.

Factors determining the morphology and distribution of astrocytes in the cat retina: A ‘contact‐spacing’ model of astrocyte interaction

The retina provides a valuable opportunity to examine the interaction of astrocytes with neurones and vasculature, in adult tissue and in vivo. We have studied astrocytes in cat retina to delineate the interactions that determine their morphology and distribution. Their morphology varied with their interaction with surrounding cells, from a classic stellate shape to an elongated bipolar form associated with axon bundles. Evidence is presented that the distribution of astrocytes across the retina is determined by their morphology and by a previously unrecognised interaction between astrocytes, which we term 'contact-spacing,' in which astrocytes maintain contact with their neighbours through their processes, but keep their somas apart. Evidence is also presented that astrocytes are not influenced in their distribution by surrounding neurones, and the influence of developmental mechanisms is identified. These observations are summarised in a contact-spacing model of astrocyte distribution, and four predictions of the model are tested. The concentration of astrocytes along axon bundles dispersed when the axons degenerate but not when vessels were prevented from forming. Further, when both axons and vessels were eliminated, the concentrations of astrocytes dispersed and they became stellate in form. Finally, in the retina of the rat, in which astrocytes show no affinity for axons, the distribution of astrocytes is essentially uniform. We suggest that the contact-spacing interaction among astrocytes provides the anatomical basis of a functional glial network extending across the retina and throughout the central nervous system.

Factors determining the migration of astrocytes into the developing retina: Migration does not depend on intact axons or patent vessels

Astrocytes migrate into the cat retina from the optic nerve, beginning from embryonic day (E) 52. Once they have entered the retina they concentrate along major axon bundles and fail to enter regions of the retina with high densities of neurones, in particular the area centralis region of the ganglion cell layer. These nonuniformities appear as the astrocytes spread over the retina during development, and in this study we have examined factors that might control their spread. First we examined astrocytes in a retina in which the axon bundles had degenerated following an optic nerve lesion at birth. The area over which astrocytes had spread was normal, suggesting that their spread does not depend on the presence of intact axons. Second, we noted that, despite the degeneration of all ganglion cells following the nerve lesion, astrocytes still did not spread over the area centralis. Their spread is apparently not inhibited by concentrations of neurones. Third, we examined astrocytes in retinas of animals raised in an atmosphere containing 70-80% oxygen, which prevents the formation of retinal vessels. Again, the area over which the astrocytes had spread was normal, suggesting that their spread does not depend on the presence of patent blood vessels. These negative findings led us to compare the distribution of spindle cells (precursors of retinal vasculature) and astrocytes in the cat during development. The close correspondence in their topographical distribution and the earlier spread of the spindle cells lead us to suggest that spindle cells provide a basal lamina component that may guide the migration of astrocytes.

“Sunbursts” in the inner plexiform layer: A spectacular feature of Müller cells in the retina of the cat

A previously unrecognised structure in the cat retina is described. Seen in Golgi-impregnated, wholemounted retinas, each such structure comprises processes radiating across the inner plexiform layer from a dense, vellate core. The processes are numerous, and largely unbranched, and give the impression of rays radiating from a point source; the structure is therefore termed a "sunburst." Evidence is presented from Golgi-impregnated retinas, and from retinas labelled with monoclonal antibodies to Müller cells, that the core of each sunburst is the inner process of a Müller cell. The sunbursts are numerous and overlap extensively, so that when neighbouring sunbursts are impregnated, they are seen to form a dense mat of processes extending across the IPL. It is suggested that each Müller cell forms a sunburst and that sunbursts form a major glial component of the neuropil of the inner plexiform layer.

Spatial characteristics of the contrast gain control in the cat's retina

Impulse rate was recorded from X- and Y-type ganglion cells in the cat's retina. The stimuli were stationary gratings for which luminance varied sinusoidally with distance across the stimulus, and amplitude varied sinusoidally in time. Y cell fundamental and second harmonic responses recorded at medium to high spatial frequencies advanced in phase with increasing contrast, an effect attributable to the contrast gain control. As spatial frequency increased to the highest value capable of evoking a second harmonic response, the phase of this response became retarded, indicating that the contrast gain control was losing its effect. This result showed that the spatial resolution of the contrast gain control is very close to that of the Y cell's rectifying subunits. The observations strongly suggest that the contrast gain control has its effect early in retinal image processing.

4B2: A monoclonal antibody to ganglion and bipolar cells in the retina of the cat, generated by intrasplenic immunization

Using tissue from the inner layers of the retina as the immunogen, we have prepared a monoclonal antibody which is selective for two classes of neuron in the cat retina. The antibody, termed 4B2, was generated by intrasplenic injection, demonstrating that neuron-specific antibodies can be elicited by this technique, which requires only micrograms of immunogen. The 4B2 binds to the somas, dendrites and axons of ganglion cells. Double labelling with 4B2 and with a retrograde tracer injected into the retino-recipient nuclei of the brain demonstrates that, of the cell classes in the ganglion cell layer, 4B2 labels only ganglion cells, and that, amongst ganglion cells, 4B2 is strongly selective for large (alpha-) and medium-sized (beta- and perhaps gamma-) cells. Double labelling with 4B2 and with markers for amacrine cells confirms tht 4B2 does not label amacrine cells. Double labelling with 4B2 and with anti-GFAP confirms that 4B2 does not label the macroglia of the retina. The 4B2 also labels bipolar cells, showing their somas, dendrites and axons. The axons of the labelled cells terminate in large boutons, in the innermost part of the inner plexiform layer and in the ganglion cell layer. This level of termination suggests that 4B2 labels rod bipolars, and perhaps a subgroup of cone bipolars. Double labelling with 4B2 and with markers for amacrine, horizontal and Müller cells indicates that other cell classes with somas in the inner nuclear layer remain unlabelled. In addition, we have examined the ontogeny of 4B2 labelling in the cat retina. The developmental sequence of labelling with 4B2 follows the sequence in which the cells are born, first ganglion and then bipolar cells.

Novel Area Serving Binocular Vision in the Retinae of Procellariiform Seabirds

Procellariiforms are pelagic seabirds which fly close to the sea surface and feed either by taking items from the surface or by shallow diving. The retinal ganglion cells in five species (Manx shearwater, Puffinus puffinus, Kerguelen petrel, Pterodroma brevirostris, great shearwater, Puffinus gravis, broad-billed prion, Pachyptila vittata, and common diving petrel, Pelecanoides urinatrix) were examined by Nissl staining and also by silver staining in the case of the common diving petrel. In all five species, a well-defined region in the dorsotemporal retina, close to the ora, was identified. This region is characterized by the presence of ganglion cells which are both regularly arrayed and larger than those found in the rest of the retina. These cells also have a large dendritic field of sparsely branched dendrites with much dendritic overlap between cells, thick axons, and dendrites confined to the proximal inner plexiform layer. Morphologically, they appear similar to the alpha cells of the retina in cats. It is suggested that the region containing these cells should be regarded as a retinal area, and the name area giganto cellularis is proposed. In the Manx shearwater, it is found that this novel area projects visually into the binocular field below the bill. Unlike previously described areas in avian retinae, it seems that this novel area is not concerned with high spatial resolution. It may function in the detection of objects on the sea surface and/or be concerned with the detection of the actual sea surface as a bird flies low over it.

TISSUE PLASMINOGEN ACTIVATOR TREATMENT OF EXPERIMENTAL SUBRETINAL HEMORRHAGE

Previous investigators have suggested that subretinal blood damages the retina in part because of its solid fibrin meshwork. The role of fibrinolysis in facilitating the clearance of subretinal hemorrhage and preventing degeneration of the overlying retina was studied. Autologous whole blood (0.1 ml) was injected into the subretinal space of 20 rabbits. Twenty-four hours later, the animals were randomized to subretinal treatment with 2.5 micrograms of tissue plasminogen activator or a similar volume of physiologic saline. Mean subretinal hemorrhage thickness 3 days after treatment had decreased to 42% of pretreatment thickness in treated eyes and remained unchanged in control eyes (P less than 0.0005). By 7 days mean clot thickness was 9% in treated eyes and 60% in controls (P = 0.005). Light microscopy revealed severe progressive retinal degeneration in both groups. No histologic evidence of retinal toxicity was found in cat retina after subretinal injection of tissue plasminogen activator (50 micrograms/ml). Although treatment with tissue plasminogen activator accelerated the clearance of subretinal hemorrhage, it failed to prevent secondary retinal degeneration in this rabbit model.

Light adaptation and frequency transfer properties of cat horizontal cells

The frequency transfer properties of horizontal cells in the cat retina were studied as a function of the mean light intensity level and stimulus contrast. To this end, horizontal cell responses to sinusoidally modulated light stimuli were recorded intracellularly in the optically intact, in vivo eye. The light stimulus consisted of a 3.9 deg dia. spot superimposed on a steady background (8.8 deg dia.). A discrete Fourier analysts was performed in order to describe the amplitude and phase characteristics of the linear response component and in order to specify the nonlinear distortion of the response. The amplitude of the fundamental Fourier component was found to increase linearly with the contrast of the sinusoidal light intensity modulation. Increasing the mean light level while keeping the contrast constant caused a frequency dependent increase in response amplitude. The increase was most pronounced at high temporal frequencies and resulted in a conspicuous increase of the flicker fusion frequency. Steady background illumination caused a reduction of the response amplitudes at the lower temporal frequencies. Responses in the high frequency range, however, were not affected. The phase shifts of the fundamental Fourier components were found to diminish at increasing mean illumination levels. The harmonic distortion of horizontal cell responses to sinewave flicker was studied as a function of stimulus frequency and stimulus contrast. By comparing the data obtained using sinusoidal light intensity modulation with the intensity profiles described in a preceding paper it was investigated to what extent the harmonic distortion can be explained by the nonlinearity expressed in the response vs intensity profiles.

Proportion of regenerating alpha and beta cells and their stratification in the cat retina

Proportion of regenerating alpha and beta cells and their stratification in the cat retina

The scotopic threshold response of the cat erg is suppressed selectively by GABA and glycine

Corneal electroretinograms (ERGs) were recorded from anesthetized cats under scotopic conditions. We examined whether the scotopic threshold response (STR) of the ERG could be functionally distinguished from scotopic PII and a-wave using intravitreal application of neuroactive agents. We found that neurotransmitters with active sites on third order neurons had several different effects. Results were: (1) glycine and γ-amino butyric acid (GABA) selectivity suppressed the STR but had relatively small and/or opposite effects on PII; (2) serotonin, acetylcholine and dopamine were nonselective and suppressed both STR and PII; (3) strychnine blocked the suppression of the STR by glycine. GABA- a antagonists alone only partially blocked GABA effects on the STR, and GABA- b antagonists were ineffective; (4) strychnine enhanced the STR. Bicuculline also increased STR amplitudes, but only in the presence of haloperidol. Our results suggest that the retinal pathway that contributes to the rod-driven STR is strongly influenced by cells that release glycine or GABA in the dark. These cells are possibly third order neurons in the retina. Our results also suggest that picrotoxin and bicuculline can facilitate the release of dopamine in the cat retina. Furthermore, the data indicate a light evoked release of dopamine which was first noticeable at about two log units above ERG threshold.

The shift effect can be elicited with both foveal and peripheral masks

Foveal target detection thresholds are elevated by presenting a counterphasing, vertical squarewave grating in the peripheral retina. This psychophysical “shift effect” has been considered to be an analogue of the neurophysiological “periphery effect” first described by McIlwain (1964; Journal of Neurophysiology, 27, 1154–1173). Physiological response properties of cells from the retina and lateral geniculate nucleus of cat and primate visual systems predict that sensitivity thresholds should also be elevated for peripheral targets in the presence of a foveal counterphasing mask. In these experiments, contrast sensitivities for human observers were obtained using a two-interval forced-choice procedure for peripheral target sinusoids in the presence of a foveal counterphasing mask. A suppressive shift effect was elicited by the foveal counterphasing squarewave mask, but only for counterphasing peripheral sinusoids. Masking was only obtained at the lowest spatial frequencies for both the peripheral and foveal shift effects.

How thick should a retina be? A comparative study of mammalian species with and without intraretinal vasculature

In this study we describe a measurement method which closely approximates in vivo retinal thickness. Using this method we examined the laminar organisation of vascular and avascular retinae from placental and marsupial mammals. Thickness measurements on retinal wholemounts show that the avascular retinae of the placental guinea pig (140 μm) and the marsupial brushtail possum (170 μm) are thinner and show less centroperipheral taper than do the vascular retinae of placental cats (250 μm), rats (220 μm) and marsupial quolls (220 μm). In general, limitation in thickness of avascular retinae is borne by most retinal layers, but most particularly by the inner plexiform layer, the synaptic region farthest removed from the choroidal blood supply. Except for the absence of blood vessels, the histological organisation of the brushtail possum's retina resembles closely that of its fellow marsupial, the quoll's. In contrast, intraretinal organisation differs amongst the two avascular retinal species with the guinea pig displaying a much coarser photoreceptor grain.

Glutamate-evoked responses in isolated bipolar cells of the cat retina

Glutamate-evoked responses in isolated bipolar cells of the cat retina

Photoreceptor degeneration and loss of immunoreactive GABA in the Abyssinian cat retina

GABA (gamma-amino butyric acid) and its synthesizing enzyme, GAD (glutamate decarboxylase; EC 4. 1. 1. 15) were localized in the retina of Abyssinian cats homozygous for a recessively inherited retinal degenerative disorder which in several respects is similar to the human disease, retinitis pigmentosa. Clinically normal mongrel cats and heterozygous Abyssinian cats were studied for comparison. The GABA and GAD immunoreactive neurons of the heterozygous or young homozygous (clinically unaffected animals) had the same distribution and morphology as normal mongrel European type cats. The neuronal GABA Immunoreactivity in both the inner and outer parts of the retina gradually disappeared in the course of the disease, with little or no loss of GAD immunoreactive neurons. Early in the disease, the changes were most severe in patches in the mid periphery of the eye and then spread both centrally and peripherally. Loss of photoreceptors was a prerequisite for the loss of GABA immunoreactivity. The observations show that retinal changes are not limited to the photoreceptors. The GABA loss is not likely to be due to a loss of neurons, because of the persistence of GAD immunoreactive neurons.

Na currents in X and W ganglion cells isolated from the adult cat retina

Na currents in X and W ganglion cells isolated from the adult cat retina

Demonstration of cell types among cone bipolar neurons of cat retina

We identified all the cone bipolar cells (80) in a small patch of one retina and then studied in detail the complete subset (42) that sends axons to sublamina b of the inner plexiform layer. The point was to learn whether the 'types' suggested previously, based on a few examples from a large population, could be substantiated or whether there would be intermediate forms. Tissue from the area centralis (1 degree eccentricity), was prepared as a series of 279 ultrathin sections and photographed in the electron microscope. Thirteen cells were reconstructed completely and parcelled into five categories (b1-b5) based on external morphology. For nine of these cells (two from categories b1-b4 and one from b5) most of the synaptic inputs and outputs were identified. When these nine cells were parcelled according to their synaptic patterns, they sorted into the same five categories. The remaining 29 cells in the population, though not reconstructed, were studied in detail by tracing their processes through the series. Ten of these cells, those near the margin of the series, were incomplete. The other 19 cells had essentially the same distribution of morphologies and synaptic patterns as the subset studied by total reconstruction: when plotted in multiparametric space, they formed distinct clusters corresponding to the five morphological categories. There was no hint of intermediate forms. That all the neurons in the population sort into some cluster (no intermediate forms), and that each neuron sorts into the same cluster by different criteria, argues that the clusters represent natural types. Each type forms a regular array in the region studied with an axonal 'coverage factor' that is close to one.