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At least 937 records · Page 52Linked to original sources
PubMed1991

Chick embryo neural retina cell culture as a screen for developmental toxicity.

An in vitro screen for developmental toxic potential of chemicals using primary cultures of chick embryo neural retina cells is described. The neural retinas of incubation Day 6.5 White Leghorn chick embryos are dissociated into single cells, which are subsequently maintained in a rotating suspension culture. Under normal circumstances, neural retina cells form spheroidal aggregates of a consistent size over the first 24 hr of culture, an event which is dependent on competent cell-cell interactions. Over the remaining 7-day period of culture, cells continue to divide and grow, and differentiation takes place. Each of these developmentally important events--aggregation, growth, and differentiation--is objectively and quantitatively measured as aggregate size and number, aggregate protein content, and glutamine synthetase (a marker of differentiation) activity, respectively. The effects on each developmental endpoint of 22 chemicals, 14 of which have been demonstrated to be developmentally toxic in one or more mammalian species in vivo, and 8 of which are not developmentally toxic, were evaluated. Chemicals were tested up to a concentration of 40 mM, or until marked cytolethality was observed. Of the known developmental toxicants, all but one, 2-methoxyethanol, affected one or more endpoints in the assay. The teratogenic metabolite of 2-methoxyethanol, 2-methoxyacetic acid, was active in the assay. None of the 8 nondevelopmental toxicants had any effect up to a concentration of 40 mM, or at biologically achievable concentrations (e.g., in vivo systemic concentrations at the LD50). Thus, the assay is 95% concordant with in vivo results for this set of chemicals. Quantitative comparisons were made (1) between developmentally toxic ip dosages in rats or mice in vivo and effective concentrations in the chick retina cell culture, and (2) between effective concentrations in chick retina cell culture and rodent whole embryo culture. In the first instance, 71% of the comparisons, and in the second instance, 89% of the comparisons, were within the same order of magnitude (and usually within a factor of two), indicating that the chick retina cell culture is also concordant with developmental toxic potency. Last, it was observed that test agents differentially affect developmental endpoints. Because the assay's endpoints are measured separately and objectively, it may be possible to use the assay to evaluate the effects of test agents on cellular development.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1982

A-type horizontal cells of the superior edge of the linear visual streak of the rabbit retina have oriented, elongated dendritic trees.

The horizontal cells of the rabbit retina have been studied by light microscopy of Golgi-impregnated whole-mount retinas. The two types of horizontal cell of the rabbit retina are similar to the horizontal cells of the cat retina in most respects. However, the majority of the A-type horizontal cells of the rabbit have asymmetrical dendritic fields compared to the circular, symmetrical dendritic fields of this cell type in the cat. The A-type horizontal cells of the superior edge of the linear visual streak in the rabbit retina are the most strikingly asymmetric and most of them are elongated and oriented in a direction approximately parallel to the linear visual streak. Like H1 axon terminals of the turtle retina the oriented, elongated A-type horizontal cells of the rabbit visual streak region may play a role in the neurocircuitry which underlies orientation sensitive ganglion cells.

Animals↗
PubMed1986

Development and degeneration of retina in rds mutant mice: effects of light on the rate of degeneration in albino and pigmented homozygous and heterozygous mutant and normal mice.

The effect of light on the rate of visual cell loss in mice afflicted by the rds (retinal degeneration slow) gene was analyzed by comparing the changes in the thickness of the outer nuclear layer. Visual cell loss in the pigmented, homozygous mutant mice, maintained in cyclic light, is slightly slower than in the albino mutant mice. In the pigmented mutant mice, exposed to constant light, and in the albino mutant mice, reared in darkness, rate of cell loss is not significantly altered. In the albino animals exposed to constant light, the rate of cell loss is faster in the homozygous mutant than in the normal, and intermediate in the heterozygous mutant retina. The accelerated cell loss in the mutant retina progresses from the centre to the periphery, and affects the rods earlier than the cones. This resembles the photic lesion in the normal retina but is unlike the genetic lesion in the mutant retina which appears to progress from the periphery to the centre and affects both rods and cones. It is concluded that the visual cells in the retina of rds mutant mice are more vulnerable to photic damage than those in the retina of normal mice.

Albinism↗
PubMed1987

Constant dendritic coverage by ganglion cells with growth of the goldfish's retina.

This study demonstrates that in the retina of the goldfish a type of ganglion cell, whose dendritic development has been well characterized [Hitchcock P.F. and Easter S. S. Jr (1986) J. Neurosci. 6, 1037-1050], is distributed across the retina in a nonrandom pattern, and the dendritic fields of this cell type overlap to completely cover, or "tile" the retina. Further, it is shown that the dendritic coverage of the retina by this cell type is established when the retina is small and is maintained as the retina grows.

Animals↗
PubMed1991

Light onset stimulates tyrosine hydroxylase activity in isolated teleost retinas.

The action of tyrosine hydroxylase is the rate-limiting step in the synthesis of dopamine, the most abundant catecholamine in vertebrate retinas. I have examined the activation and regulation of this enzyme in isolated retinas of green sunfish, Lepomis cyanellus. Exposing previously dark-adapted retinas to constant illumination for a period of 10 min increased enzymatic activity 2.2-fold over that present in retinas incubated in darkness. Thus, light onset activates tyrosine hydroxylase in teleost retinas. Stimulation of the activity of tyrosine hydroxylase under these conditions was associated with a decrease in the apparent Km of the enzyme for its pteridine cofactor without a change in the apparent Vmax of the reaction. This result suggests that short-term exposure to light increases dopamine synthesis by enhancing the affinity of the enzyme for its naturally occurring cofactor. These findings are consistent with the idea that light activates dopaminergic neurons in teleost retinas.

Animals↗
PubMed1992

Neurons immunoreactive to choline acetyltransferase in the turtle retina.

Light microscopic immunocytochemistry using anti-choline acetyltransferase (ChAT) was performed to stain putative cholinergic amacrine cells in turtle retina. ChAT-immunoreactive somata lie in the inner nuclear (INL) and ganglion cell (GCL) layers. Three types of amacrine cells were found according to the location of their somata and their dendritic stratification pattern in the inner plexiform layer (IPL). Type I amacrines lie in the row of cells closest to the INL/IPL limits and they branch along the s1/s2 border of the IPL. Type II amacrines are displaced to the GCL and they ramify along the s3/s4 border of the IPL. Type III amacrines lie in the middle of the INL, 2-3 rows away from the IPL limits and their dendrites appear to be bi- or tri-stratified in s1 and s3-s4 of the IPL. The turtle ChAT-IR amacrines are thus similar to the types described in chicken retina. A regular, non-random mosaic formed by stained type II amacrine cells was observed in the GCL. Their density in mid-central retina was 750 cells/mm2, tapering off to 393 cells/mm2 in peripheral retina. Our study indicates that a pair of cholinergic amacrine cell types in turtle retina is arranged in mirror-image symmetry contributing to sublamina "a" and sublamina "b" of the IPL, like in other vertebrate retinas.

Animals↗
PubMed1993

The spatial organization of tyrosine hydroxylase-immunoreactive amacrine cells in the chicken retina and the consequences of myopia.

We examined the spatial organization of the putative dopaminergic amacrine cells in the chicken retina and how this organization was affected by myopic eye enlargement. Myopia was produced by monocular lid suture for 4-7 months from hatching. Dopaminergic amacrine cells (TH-IR) were labelled by tyrosine hydroxylase immunohistochemistry. The somata of the TH-IR cells were usually located at the inner border of the inner nuclear layer; they gave rise to a dense plexus in stratum 1 (S1) of the inner plexiform layer, to a sparse plexus in stratum 3 (S3), and to short spiny dendrites at the border of strata 4 and 5 (S4/S5). The long thin processes in S1 and S3 could seldom be traced to their cell of origin, whereas the S4/S5 dendrites formed discrete fields that tiled the retina with little overlap. Lid suture resulted in retinal expansion of between 25-70%, but the total number of TH-IR amacrine cells was unaltered. Per retina, there were about 4700 TH-IR amacrine cells which showed a 3:1 density gradient from central to peripheral retina. The size of the S4/S5 dendritic fields increased proportionately in the expanded retinae, thus maintaining their coverage across the retina. The increase was achieved through scaled growth of the S4/S5 dendrites, involving both terminal and non-terminal dendrites. These findings suggest that the expansion of retinal neurons during myopia occurred through normal, albeit excessive, growth mechanisms.

Animals↗
PubMed1994

Co-localization of GABA- and tyrosine hydroxylase-like immunoreactivities in amacrine cells of the rabbit retina.

Previous studies have shown that many of the tyrosine hydroxylase-immunoreactive (TH-IR) amacrine cells in the retina of mammals also show GABA-like immunoreactivity (GABA-IR). However such co-localization has yet to be demonstrated in the rabbit retina. In this study the proportion of TH-IR amacrine cells that show GABA-IR has been determined in the retina of the cat, rat and rabbit following fixation with 4% paraformaldehyde plus varying concentrations of glutaraldehyde. In the retina of the cat and rat, most of the TH-IR amacrine cells showed GABA-IR following fixation using low concentrations (0.01 or 0.1%) of glutaraldehyde. However in the rabbit retina very few of the TH-IR amacrine cells showed GABA-IR unless the glutaraldehyde concentration in the fixative was 0.5% or greater. The results suggest that the dopaminergic amacrine cells in the rabbit retina contain only very low levels of GABA, which are close to the detection limit of the antibody.

Animals↗
PubMed1994

Immunocytochemical characterization and spatial distribution of midget bipolar cells in the macaque monkey retina.

Midget bipolar cells form the first distinct step in the parvocellular (P-) pathway of the primate visual system, and are the major determinant of the receptive field properties of colour selective midget ganglion cells. This paper describes the sampling properties of the midget bipolar cell population and relates this to the processing of chromatic information in the P-pathway. Immunocytochemical markers were used to label midget bipolar cells so that their spatial density could be compared with that of cones and ganglion cells. Sections through macaque monkey retinae were immunostained with antibodies against cholecystokinin (CCK), and recoverin. In CCK-labelled sections, in addition to blue cone bipolar cells, numerous thin bipolar cell dendrites, which could be associated with individual cone pedicles are stained. CCK-immunoreactive midget bipolar cells are found throughout the retina. A different population of midget bipolar cells is revealed in recoverin-labelled sections. Based on a comparison with midget bipolar cells in Golgi-stained retinae we propose that ON-midget (invaginating) bipolars are immunoreactive for CCK and confirm that OFF-midget (flat) bipolar cells are immunoreactive for recoverin [Milam, Dacey and Dizhoor (1993) Visual Neuroscience, 10, 1-12]. The density of recoverin labelled midget bipolars matches the cone density to an eccentricity of about 10 mm; from there outwards it drops to 60% of the cone density. This suggests convergence of several cones to individual midget bipolar cells in peripheral retina. We conclude that midget bipolar cells are present throughout the entire primate retina, and could, in peripheral as well as in central retina, provide chromatically specific input to the P-pathway.

Animals↗
PubMed1976

Stimulation of DNA synthesis by ouabain and concanavalin A in cultures of embryonic neural retina cells.

Ouabain and concanavalin A, agents which bind to specific sites in the cell membrane, stimulate DNA synthesis and cell replication in monolayer cultures of neural retina cells from late chick embryos. The results suggest a relationship between control of retina cell replication and properties of the cell membrane. The experiments involved measurements of 3H-thymidine incorporation in primary monolayer cultures (24-48h) of retina cells from embryos of different ages. Stimulation by ouabain was greatest in cells from 14-day embryos, and its magnitude was similar to that elicited in these cell cultures by concanavalin A. Simultaneous treatment of 14-day retina cells with both agents resulted in a greater than additive stimulation of DNA synthesis. Our results demonstrated that, although during normal embryogenesis cell replication in the neural retina has virtually ceased by day 14 of development, some cells retained a capacity for mitogenesis when exposed to conditions such as provided in these experiments. By autoradiography the responding cells were identified as large epithelioid retina cells (LER cells). Under optimal conditions of simultaneous treatment with ouabain and Con A about 20% of the LER cells showed stimulation of DNA synthesis. The nature of LER cells and other aspects of our findings are discussed.

Animals↗
PubMed1987

Lipofuscin autofluorescence: evidence for vitamin A involvement in the retina.

A lipofuscin-like autofluorescence develops in the degenerating photoreceptor cells of the RCS rat, a strain with inherited retinal dystrophy. In animals with normal retinas, age-related lipofuscin accumulation in the eye is restricted to the retinal pigment epithelium (RPE). Previous investigations have established that RPE lipofuscin accumulation in the normal rat retina can be reduced by dietary vitamin A deficiency. In order to determine whether the photoreceptor-derived fluorescence in the RCS rat retina is related to RPE lipofuscin fluorescence, the influence of dietary vitamin A on the fluorophore content of the RCS rat retina was studied. Vitamin A deficiency substantially reduced the autofluorescence associated with degenerating photoreceptor cells of the RCS rat retina. A specific vitamin A-dependent fluorophore was isolated from these retinas using thin-layer chromatography (TLC). The mobility of this fluorophore on TLC differs from that of the major age-dependent fluorophore isolated from the RPE of normal rats. Thus, if the vitamin A-dependent fluorophores of the photoreceptors and RPE are related, it appears that the fluorophore generated in the photoreceptor cells must undergo chemical modification once it has been taken up by the RPE. The fact that both the RPE- and photoreceptor-associated fluorophores are vitamin A-dependent suggests that such a relationship between them is likely. These experiments indicate that the RPE is somewhat different from other lipofuscin-accumulating tissues in that a major precursor of RPE lipofuscin fluorophores originates in another cell type and enters the RPE via phagocytosis.

Animals↗
PubMed1989

An eyecup slice preparation for intracellular recording in vertebrate retinas.

This report describes a new preparation for intracellular recording from the vertebrate retina, the eyecup slice preparation. It consists of a small (2 X 5 mm) strip cut from the posterior wall of the eye and thereby keeps the sclera, pigment epithelium and neural retina in place. Initial results are presented here for two vertebrates: the turtle, Pseudemys scripta elegans, and the toad, Bufo marinus. With conventional microscopy, the histological layers of the retina can be resolved, as well as individual photoreceptors, to provide landmarks for intracellular recording. When superfused, the eyecup slice remains in good condition for many hours and yields intracellular recordings of good quality and stability. Recordings of the light-evoked responses of cones and horizontal cells show that the slice is large enough to preserve the characteristic spatial interactions mediated by the laterally coursing neural networks of the distal retina. Recordings from rods show that full dark adaptation is achieved. Thus, photochemical dark adaptation as well as other normal cellular interactions between the neural retina and pigment epithelium can be preserved in this preparation, in contrast to isolated retinal slice preparations. The eyecup slice preparation might be particularly useful for work on mammalian retinas.

Adaptation, Physiological↗
PubMed1995

A novel, rapid flat-mounting technique for visualizing antibody labeling in the retina.

Complete analysis of retinal tissue is difficult because it consists of a thin neural tissue spread across the back of a hemispheric surface. Conventional sectioning in a plane parallel to a central axis of symmetry produces a large number of samples, each containing only a small amount of the tissue of interest. Consequently, quantitative comparison of any feature of interest typically uses a small fraction of the sections from each retina, because analysis of the entire collection of sections is too time consuming. Such a sampling process can lead to misleading or erroneous conclusions. We present a new method which allows complete analysis of the retina using a small number of samples produced by sectioning flattened retinas. This procedure is straightforward as illustrated using an antibody against proliferating cell nuclear antigen (PCNA) to locate dividing cells in the teleost fish retina. Immunocytochemical staining on flat-sectioned retinas was quantified using a computer-based image analysis system. When the cells of interest are randomly distributed, conventional sampling procedures can seriously under- or over-estimate their number. The new technique presented allows significantly more efficient examination and quantification of the entire retina as compared to conventional techniques.

Animals↗
PubMed1983

Lucifer yellow uptake in developing rat retina: selective staining of horizontal cells.

We have examined the uptake of Lucifer yellow CH by developing rat retina. We find that selective dye uptake occurs in retina incubated in vitro in the absence of external Ca2+. The pattern of uptake appears to depend on the developmental stage examined. Although little uptake is seen before postnatal day 5, between days 6 and 9, the dye selectively stains putative horizontal cell bodies and their process. Later, other cell bodies located deeper in the INL begin to show staining until day 21, when the uptake pattern becomes analogous to that seen in adult retina. By nearest neighbor analysis of labeled cell bodies in wholemounts of D-9 retina, we have shown that the labeled cells are distributed in a quasi-regular mosaic in the retina. Upon proteolytic dissociation of labeled retina, individual cells with Lucifer yellow fluorescence could be readily identified in the cell suspensions.

Animals↗
PubMed1984

In vitro effects of kainate on embryonic and posthatching chick retina.

The onset and developmental pattern of kainate effects were determined in isolated embryonic and posthatching chick neural retinas exposed to KA (0.01-0.24 mM; 1-30 min) in vitro. Damage was first apparent in day 8 embryonic retinas; cells in the inner region of the inner nuclear layer and neuronal processes in the inner plexiform layer were affected. In older embryonic retinas, cells in the inner region of the inner nuclear layer continued to be the most damaged although cells in the ganglion cell layer and the outer region of the inner nuclear layer, and neurites in the outer plexiform layer were also affected. Cells in the outer nuclear layer stained normally in both embryonic and posthatching retinas. Partial reversibility of kainate effects was observed in retinas which had been exposed briefly to kainate and then allowed to recover in kainate-free medium. The results of our in vitro experiments are consistent with those reported for a variety of neonatal and adult retinas treated with kainate in vivo.

Animals↗
PubMed1985

Gangliosides in postmitotic retina of chick embryo: changes in vivo and in cell cultures.

Developmental changes in ganglioside composition of postmitotic neural retina of chick embryo were analyzed by thin-layer chromatography. Gangliosides were identified by comparing their chromatographic mobilities with reference standards. The outstanding changes are decrease in the concentration of GD3L and increase in GD1a and GM1 concentrations. By depleting Müller glia cells from retina tissue of 13- and 16-day embryos (R13, R16) we determined that the bulk of the major gangliosides is associated with the neurons. Analysis of gangliosides in monolayer cultures of R13 and R16 cells highly enriched for Müller cell-derived gliocytes indicated that these cells express the same types of gangliosides as neurons, but in somewhat different concentrations and relative proportions; however, after time in culture these cells showed ganglioside types and changes in ganglioside profile that are not characteristic of normal retina. The latter observation is consistent with other evidence that the phenotype of Müller glia cells becomes altered in monolayer culture. In contrast to cultures of early embryonic retina, in organ cultures of later postmitotic retina, ganglioside composition did not continue to change as in normal development. This suggests that in postmitotic retina, normal developmental progression of ganglioside changes requires systemic and/or other conditions which are missing or altered when this tissue is isolated and cultured in vitro.

2-Aminoadipic Acid↗
PubMed1996

Calbindin and parvalbumin immunoreactivity in the developing and adult human retina.

We report the expression and pattern of two calcium-binding proteins (CBPs), calbindin (CALB) and parvalbumin (PV), in the fetal (13-25 weeks of gestation), postnatal (5 months) and adult human retina, as studied by immunohistochemistry. Both CBPs appear prenatally in different neurones as well as in the nerve fibre, inner and outer plexiform layers. The cones do not show immunoreactivity for both CBPs up to 25 weeks, the last fetal stage studied; however, they are immunopositive in the postnatal and adult retina. Of the two CBPs, CALB appears first, followed by PV. The immunoreactivity for both CALB and PV in the fetal retina follows a centroperipheral gradient and vitreal to scleral pattern of retinal differentiation. The CALB immunoreactivity shows a gradual increase in intensity with age. A spurt in intensity of PV immunoreactivity in the 24-25 week fetal retina and increased intensity in the 60 year normal adult retina when compared to the light-deprived retina of a 34-year-old staphyloma patient suggests an activity-dependent functional role for PV.

Adult↗
PubMed1993

Fibroblast growth factor induces proliferating cell nuclear antigen-immunoreactive cells in goldfish retina.

New rod photoreceptors are added to mature teleost retinas throughout life by regulated proliferation of rod precursor cells (RPCs). In this study, candidate regulators of RPC proliferation, acidic and basic fibroblast growth factors (aFGF and bFGF; 0.1 microgram/eye), interleukin-6 (IL-6; 0.1 microgram) and phytohaemagglutinin (HA15; 1.0 microgram), were injected intravitreally into one eye of goldfish (body length 5-6 cm), and mitotic RPCs in both retinas were detected and counted 3-50 days later by immunohistochemistry for proliferating cell nuclear antigen (PCNA). Retinal integrity after treatment was assessed by immunohistochemistry for tyrosine hydroxylase (TH) and other retinal antigens. All the agents applied altered the density of PCNA-immunoreactive (ir) cells in the outer and inner nuclear layers (ONL and INL) in both retinas as soon as 2-3 days after unilateral injection. Initially (2-20 days after injection), particularly in the treated retina, PCNA-ir cells appeared in clusters accompanied by various numbers of scattered individual cells, but subsequently the clusters of PCNA-ir cells disappeared while the density of singly distributed cells increased until 30 days after injection. At the doses given, these effects were most striking with aFGF and bFGF and less with IL-6 and HA15. In radial cryosections, other cellular elements immunoreactive to markers such as TH, serotonin, neuropeptide Y, substance P, glutamine synthetase, glial fibrillary acidic protein and protein kinase C, were found normal in terms of morphology. In addition, a monoclonal antibody (NN-2) was found to label some non-neuronal structures (macrophages, microglia and blood vessels) inside and outside the retina intoxicated with 6-hydroxydopamine, a few NN-2-ir cells being PCNA-positive. However, clustered PCNA-ir and marginal neuroblast cells were NN-2-negative. These results indicate that FGFs may play an important role in stimulating the proliferation of RPCs, for example, in the regeneration of fish retinas following neurotoxic destruction.

Animals↗