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PubMed1993

Developmental control of glucocorticoid receptor transcriptional activity in embryonic retina.

In chicken embryo retina, competence for induction of the glutamine synthetase [L-glutamate:ammonia ligase (ADP-forming); EC 6.3.1.2] gene by glucocorticoid hormones increases progressively with development; this competence is minimal in 6-day retina (E6) and high by day 10 (E10). Because the level of glucocorticoid receptors (GRs) in the retina does not increase during that time, we investigated whether the transcriptional activity of GR increased between days 6 and 10 of development. The glucocorticoid-inducible chloramphenicol acetyltransferase (CAT) constructs 2GRE-37TK and p delta G46TCO, which contain glucocorticoid-responsive elements attached to a TATA box and to the thymidine kinase promoter, respectively, were transfected into E6 and E10 retinas, and their inducibility was examined. CAT expression could be induced in the transfected E10 retina but was not induced in the transfected E6 retina. However, induction was obtained also in E6 retina after cotransfection with a GR expression vector. Noninducible CAT constructs (pRSV-CAT, pSV2CAT, and pBLCAT2) were expressed at both ages at similar levels. The CAT construct pGS2.1CAT, which is controlled by the upstream sequence of the chicken glutamine synthetase gene, could be induced in E10 retina but was not induced in E6 retina; however, cotransfection with the GR expression vector resulted in induction of pGS2.1CAT also in E6 retina. We interpret these results as showing that the transcriptional activity of GR in embryonic retina is developmentally controlled and suggest that its increase is causally implicated in the development of competence for glutamine synthetase induction.

Animals↗
PubMed1979

Autoradiographic identification of acetylcholine in the rabbit retina.

Rabbit retinas were studied in vitro under conditions known to maintain their physiological function. Retinas incubated in the presence of [3H]choline synthesized substantial amounts of both [3H]phosphorylcholine and [3H]acetylcholine. With time, [3H]phosphorylcholine proceeded into phospholipids, primarily phosphatidylcholine. Retinas pulse-labeled by a 15-min exposure to 0.3 microM [3H]choline were incubated for a subsequent hour under chase conditions designed either to retain newly synthesized acetylcholine within synapses or to promote its release. At the end of this time the two groups of retinas were found to contain equal amounts of radioactivity in the phospholipid pathway, but only the retinas incubated under the acetylcholine-protecting conditions contained [3H]acetylcholine. Freeze-dried, vacuum-embedded tissue from each retina was autoradiographed on dry emulsion. All retinas showed silver grains over the photoreceptor cells and faint labeling of all ganglion cells. In the retinas that contained [3H]acetylcholine, silver grains also accumulated densely over a few cells with the position of amacrine cells, over a subset of the cells of the ganglion cell layer, and in two bands over the inner plexiform layer. Fixation of the retina with aqueous osmium tetroxide retained only the radioactive compounds located in the photoreceptor and ganglion cells. Sections from freeze-dried tissue lost their water-soluble choline metabolites when exposed to water, and autoradiography of such sections again revealed radioactivity primarily in the photoreceptor and ganglion cells. Radioactive compounds extracted from the sections were found to faithfully reflect those present in the tissue before processing; analysis of the compounds eluted from sections microdissected along the outer plexiform layer showed [3H]acetylcholine to have been synthesized only by cells of the inner retina. Taken together, these results indicate that the photoreceptor and ganglion cells are distinguished by a rapid synthesis of choline-containing phospholipids, while acetylcholine synthesis is restricted to a few cells at both margins of the inner plexiform layer. They imply that the only neurons to release acetylcholine within the rabbit retina are a small group of probable amacrine cells.

Acetylcholine↗
PubMed1989

Light-induced dopamine release from teleost retinas acts as a light-adaptive signal to the retinal pigment epithelium.

In the retinal pigment epithelium (RPE) of lower vertebrates, melanin pigment granules migrate in and out of the cells' long apical projections in response to changes in light condition. When the RPE is in its normal association with the retina, light onset induces pigment granules to disperse into the apical projections; dark onset induces pigment granules to aggregate into the cell bodies. However, when the RPE is separated from the retina, pigment granule movement in the isolated RPE is insensitive to light onset. It thus seems likely that a signal from the retina communicates light onset to the RPE to initiate pigment dispersion. We have examined the nature of this retina-to-RPE signal in green sunfish, Lepomis cyanellus. In isolated retinas with adherent RPE, light-induced pigment dispersion in the RPE is blocked by treatments known to block Ca2+-dependent transmitter release in the retina. In addition, the medium obtained from incubating previously dark-adapted retinas in the light induces light-adaptive pigment dispersion when added to isolated RPE. In contrast, the medium obtained from incubating dark-adapted retinas in constant darkness does not affect pigment distribution when added to isolated RPE. These results are consistent with the idea that RPE pigment dispersion is triggered by a substance that diffuses from the retina at light onset. The capacity of the conditioned medium from light-incubated retinas to induce pigment dispersion in isolated RPE is inhibited by a D2 dopamine antagonist, but not by D1 or alpha-adrenergic antagonists. Light-induced pigment dispersion in whole RPE-retinas is also blocked by a D2 dopamine antagonist.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Topographical analysis of peripheral vs central retina with pattern reversal visual evoked response and the scanning laser ophthalmoscope.

The contribution of the central and the surrounding peripheral retina to the pattern reversal visual evoked response (PVER) was analyzed simultaneously using double frequency stimulation and the Fast Fourier transform (FFT) method. The Scanning Laser Ophthalmoscope (SLO) was used to project the pattern stimulus, a square subtending 10 degrees on a side, on a specific location in the fundus and to monitor accurately the stimulus' position during PVER testing. When the stimulus area on the central retina was less than or equal to 50 min of arc on a side, or 0.69% of the total area was stimulated (a square of 10 degrees on a side), the contribution from the central retina was negligible, and the PVER was dominated by surrounding peripheral retinal activity. When the stimulus area of the central retina was greater than or equal to 100 min of arc on a side, or 2.78% of the whole area was stimulated, activity from the central retina became more evident. When the stimulus area projected on the central retina became 150 min on a side (6.25% of the area or greater), activity from the central retina showed an increase and exceeded that of the peripheral retina. When the central area became larger, its activity dominated the PVER, and the contribution from the peripheral retina became negligible at this stage. The PVER was highly dependent upon the activity of the central retina, and activity levels from the central and surrounding peripheral retina were correlated.

Adult↗
PubMed2006

The iron carrier transferrin is upregulated in retinas from patients with age-related macular degeneration.

PURPOSE: Iron can cause oxidative stress, and elevated iron levels have been associated with several neurodegenerative diseases including age-related macular degeneration (AMD). Transferrin, an iron transport protein, is expressed at high levels in the retina. The purpose of this study was to assess transferrin involvement in AMD by determining the expression profile of transferrin in retinas with AMD compared with retinas without evidence of disease. METHODS: Postmortem retinas were obtained from AMD and non-AMD eyes. Expression of transferrin was assessed in a microarray dataset from 33 retinas of unaffected donors and 12 retinas of patients with AMD (six with neovascular AMD and six with non-neovascular AMD). Quantitative real-time RT-PCR (QPCR) was used to confirm the microarray results. Transferrin protein expression was assessed by semiquantitative Western blot analysis and immunohistochemistry. RESULTS: In comparison to unaffected retinas, mean transferrin mRNA levels, as measured by microarray analysis were elevated 3.5- and 2.1-fold in non-neovascular and neovascular AMD retinas, respectively. Semiquantitative Western blot analysis demonstrated a 2.1-fold increase in transferrin protein in AMD eyes. Immunohistochemistry showed more intense and widespread transferrin label in AMD maculas, particularly in large drusen, Müller cells, and photoreceptors. CONCLUSIONS: These data demonstrate that transferrin expression is increased in the retinas of patients with AMD relative to those of healthy control patients of comparable age. Along with previous studies that have demonstrated elevated iron levels in AMD retinas, early onset drusen formation in a patient with retinal iron overload resulting from aceruloplasminemia, and retinal degeneration with some features of macular degeneration in the iron-overloaded retinas of ceruloplasmin/hephestin knockout mice, the present study suggests that altered iron homeostasis is associated with AMD.

Adult↗
PubMed2005

Comprehensive in silico functional specification of mouse retina transcripts.

BACKGROUND: The retina is a well-defined portion of the central nervous system (CNS) that has been used as a model for CNS development and function studies. The full specification of transcripts in an individual tissue or cell type, like retina, can greatly aid the understanding of the control of cell differentiation and cell function. In this study, we have integrated computational bioinformatics and microarray experimental approaches to classify the tissue specificity and developmental distribution of mouse retina transcripts. RESULTS: We have classified a set of retina-specific genes using sequence-based screening integrated with computational and retina tissue-specific microarray approaches. 33,737 non-redundant sequences were identified as retina transcript clusters (RTCs) from more than 81,000 mouse retina ESTs. We estimate that about 19,000 to 20,000 genes might express in mouse retina from embryonic to adult stages. 39.1% of the RTCs are not covered by 60,770 RIKEN full-length cDNAs. Through comparison with 2 million mouse ESTs, spectra of neural, retinal, late-generated retinal, and photoreceptor -enriched RTCs have been generated. More than 70% of these RTCs have data from biological experiments confirming their tissue-specific expression pattern. The highest-grade retina-enriched pool covered almost all the known genes encoding proteins involved in photo-transduction. CONCLUSION: This study provides a comprehensive mouse retina transcript profile for further gene discovery in retina and suggests that tissue-specific transcripts contribute substantially to the whole transcriptome.

Animals↗
PubMed1995

[Age-related changes in the retina of WBN/Kob rats--a pathological study].

Male rats of the WBN/Kob strain, which are known to spontaneously develop diabetes with aging, were examined for histopathological changes in the retina. Five rats (10 eyes) each of WBN/Kob and Wistar/ST as a control were used, and the thickness of the retinal layers, both the central region and the peripheral region of the retina, were measured on weeks 1, 2, 3, 5, 9, 13, 17, 23, 27, 36, 45, 54, 67 and 80 after birth. The rod and cone cell layer in WBN/Kob rats was under-grown, and its thickness decreased 71.7% in the central zone and 59.3% in the peripheral zone of the retina compared with that of the control. In the central and peripheral retina, the rod and cone cell layer, outer nuclear layer and outer plexiform layer, as well as the inner plexiform layer in the central retina, gradually decreased in thickness from 5-45 weeks of age. In the central and peripheral parts of the retina, the number of nuclei decreased in accordance with the thinning of the nuclear layer. The thinned layers showed only cell loss. The rod and cone cell layer of the peripheral retina was thinner than that of the central retina. We obtained the following findings in the retina of male WBN/Kob rats. First, the rod and cone cell layer is undergrown compared with that of the control. Second, the first change occurred in the rod and cone cell layer. Third, the thinning of the rod and cone cell layer appeared at 5 weeks of age, and thinning with aging was slow. And finally, the thinning of the peripheral retina was more severe than that of the central retina. From the above findings, it seems that retinal changes in WBN/Kob rats are similar to the retinal degeneration of rds mice (retinal degeneration slow mice) and that WBN/Kob rats provide a useful animal model for human retinopathy.

Aging↗
PubMed2001

IGF-I mRNA and signaling in the diabetic retina.

IGF-I promotes the survival of multiple cell types by activating the IGF-I receptor (IGF-IR), which signals downstream to a serine/threonine kinase termed Akt. Because in diabetes vascular and neural cells of the retina undergo accelerated apoptosis, we examined IGF-I synthesis and signaling in the human and rat diabetic retina. In retinas obtained postmortem from six donors aged 64 +/- 8 years with a diabetes duration of 7 +/- 5 years, IGF-I mRNA levels were threefold lower than in the retinas of six age-matched nondiabetic donors (P = 0.005). In the retinas of rats with 2 months' duration of streptozotocin-induced diabetes, IGF-I mRNA levels were similar to those of control rats, but after 5 months of diabetes they failed to increase to the levels recorded in age-matched controls (P < 0.02). Retinal IGF-I expression was not altered by hypophysectomy, proving to be growth-hormone independent. IGF-IR levels were modestly increased in the human diabetic retinas (P = 0.02 vs. nondiabetic retinas) and were unchanged in the diabetic rats. Phosphorylation of the IGF-IR could be measured only in the rat retina, and was not decreased in the diabetic rats (94 +/- 18% of control values). In the same diabetic rats, phosphorylation of Akt was 123 +/- 21% of control values. There was not yet evidence of increased apoptosis of retinal microvascular cells after 5 months of streptozotocin-induced diabetes. Hence, in the retina of diabetic rats, as in the retina of diabetic human donors, IGF-I mRNA levels are substantially lower than in age-matched nondiabetic controls, whereas IGF-IR activation and signaling are not affected, at least for some time. This finding suggests that in the diabetic retina, the activation of the IGF-IR is modulated by influences that compensate for, or are compensated by, decreased IGF-I synthesis.

Aged↗
PubMed2000

Morphologic preservation and variability of human donor retina.

PURPOSE: To facilitate studies of human retina and utilization of human retinal tissue for treatment of retinal diseases, we studied morphologic preservation in postmortem human retina. METHODS: Morphology of retinas from thirty-one human eyes was examined using light and electron microscopy. The inner and outer retina, rod and cone photoreceptor cells, and central and peripheral retina were compared with regard to morphologic preservation. Possible factors affecting survival were analyzed. RESULTS: The earliest postmortem change was vacuolation of the nerve fiber layer within a few hours postmortem, followed by vacuolation and cytoplasmic swelling of the inner retina. As compared with the inner retina, outer retinal structure was better preserved, i.e., the photoreceptor cells maintained better morphology. Rod cell morphology was better preserved than cone cell morphology, with good preservation of the rod outer segment disc membranes and the inner segment mitochondrial membranes. Thus, well-preserved rod photoreceptor cells were evident in specimens at least 48-hours postmortem. Peripheral retina was better preserved than the central retina including the fovea and perifovea. Factors affecting anatomical integrity included the total time postmortem and, more importantly, the time between death and enucleation. Other factors, including age and sex, did not appear to affect morphological preservation in the present study. CONCLUSIONS: Human retina postmortem remained morphologically intact for a relatively long period of time, with differential preservation among different geographic areas and cell types. This morphologic evidence is consistent with previous findings of functional preservation (e.g. , photoresponses) in such tissue. This study may shed some light on understanding of human retina and its utilization for retinal transplantation.

Adolescent↗
PubMed2000

Lutein and zeaxanthin concentrations in rod outer segment membranes from perifoveal and peripheral human retina.

PURPOSE: In addition to acting as an optical filter, macular (carotenoid) pigment has been hypothesized to function as an antioxidant in the human retina by inhibiting the peroxidation of long-chain polyunsaturated fatty acids. However, at its location of highest density in the inner (prereceptoral) layers of the foveal retina, a specific requirement for antioxidant protection would not be predicted. The purpose of this study was to determine whether lutein and zeaxanthin, the major carotenoids comprising the macular pigment, are present in rod outer segment (ROS) membranes where the concentration of long-chain polyunsaturated fatty acids, and susceptibility to oxidation, is highest. METHODS: Retinas from human donor eyes were dissected to obtain two regions: an annular ring of 1.5- to 4-mm eccentricity representing the area centralis excluding the fovea (perifoveal retina) and the remaining retina outside this region (peripheral retina). ROS and residual (ROS-depleted) retinal membranes were isolated from these regions by differential centrifugation and their purity checked by polyacrylamide gel electrophoresis and fatty acid analysis. Lutein and zeaxanthin were analyzed by high-performance liquid chromatography and their concentrations expressed relative to membrane protein. Preparation of membranes and analysis of carotenoids were performed in parallel on bovine retinas for comparison to a nonprimate species. Carotenoid concentrations were also determined for retinal pigment epithelium harvested from human eyes. RESULTS: ROS membranes prepared from perifoveal and peripheral regions of human retina were found to be of high purity as indicated by the presence of a dense opsin band on protein gels. Fatty acid analysis of human ROS membranes showed a characteristic enrichment of docosahexaenoic acid relative to residual membranes. Membranes prepared from bovine retinas had protein profiles and fatty acid composition similar to those from human retinas. Carotenoid analysis showed that lutein and zeaxanthin were present in ROS and residual human retinal membranes. The combined concentration of lutein plus zeaxanthin was 70% higher in human ROS than in residual membranes. Lutein plus zeaxanthin in human ROS membranes was 2.7 times more concentrated in the perifoveal than the peripheral retinal region. Lutein and zeaxanthin were consistently detected in human retinal pigment epithelium at relatively low concentrations. CONCLUSIONS: The presence of lutein and zeaxanthin in human ROS membranes raises the possibility that they function as antioxidants in this cell compartment. The finding of a higher concentration of these carotenoids in ROS of the perifoveal retina lends support to their proposed protective role in age-related macular degeneration.

Aged↗
PubMed2000

Localization and characterization of immunocompetent cells in the human retina.

PURPOSE: Recent studies have shown that experimental uveitis can be induced by the appropriate administration of various retinal antigens. Little is known about the in-situ interactions between immune cells in the retina as a prerequisite for understanding the mechanisms involving the presentation of antigens by local antigen-presenting cells (APC) to invading T cells. The study described here was therefore designed to investigate the presence of immunocompetent cells with a focus on the characterization of retinal APCs. MATERIALS AND METHODS: Retinal wholemounts, cytospins, and ocular sections were prepared from eyebank eyes obtained within 24 hours postmortem. Immunohistochemistry (single staining and double staining) was performed on the retinal wholemounts, cytospins, and the ocular sections using monoclonal antibodies specific for HLA-DR (MHC class II), CD45 (leukocytes), CD68 (macrophages), CD22 (B cells), CD3 (T cells), and CD1a (dendritic cells). RESULTS: CD68-positive macrophages were observed in one layer of the retina, whereas HLA-DR(+) and CD45(+) cells were seen in two distinct planes: one mainly at the level of the inner nuclear layer to outer plexiform layer (deep layer) and the other mainly at the nerve fiber and ganglion cell layer (shallow layer). There was a significant difference between the different parts of the retina with regard to the density of these cells. Cell density decreased when going from the peripheral to the posterior areas of the retina. The positive cells in the deep layer were frequently associated with blood vessels, whereas the cells in the shallow layer were distributed evenly throughout the retina. Most positive cells displayed a dendritiform appearance, whereas few cells showed a pleiomorphic morphology. Very few CD1a-positive cells were noted in the retina. Neither T cells (CD3) nor B cells (CD22) could be detected in the normal human retina. Double staining showed that the majority (83. 7%) of the CD45(+) cell population was HLA-DR-positive, whereas approximately half (56.8%) the CD68(+) cell population was HLA-DR-positive. CONCLUSIONS: This study shows that the human retina contains a number of different microglia populations, some of which express HLA-DR and could thus be involved in antigen presentation. Marked differences in cell density can be observed within the retina, with the most abundant presence seen in the peripheral retina. The normal retina contains few professional antigen-presenting cells (CD1(+)).

Adult↗
PubMed2003

Expression of cytokine signal transduction components in the postnatal mouse retina.

PURPOSE: Members of the ciliary neurotrophic factor (CNTF) family of cytokines have been shown to influence neuronal differentiation during retinal development and enhance cell survival in various retinal degeneration models. However, the cellular mechanism of CNTF signaling and the target cell types for CNTF in the developing retina remain unidentified. The purpose of this study is to characterize expression patterns of proteins involved in cytokine signal transduction in the mouse retina, thus to assess the potential responsiveness of different retinal cell types to CNTF-like cytokine signals. METHODS: The expression profiles of various cytokine signal transduction components, including receptor subunits CNTF receptor alpha (CNTFRa) and gp130, intracellular protein kinases, Jak2 and Tyk2, as well as latent transcription factors, STAT1 and STAT3, were determined by immunohistochemical staining of mouse retinal sections derived from different postnatal stages. In addition, the distribution of ERK was studied by immunofluorescent staining. RESULTS: In the neonatal retina, intense staining signals for gp130, CNTFRalpha, Jak2, Tyk2, STAT1, and STAT3 were present in the differentiated ganglion cell layer and the developing inner plexiform layer of the mouse retina. Detectable staining signals were also observed in the ventricular zone of the early postnatal mouse retina. From P5 to P10, cytokine signaling molecules also accumulated in the developing outer plexiform layer. In the adult retina, cytokine signaling components examined were localized to the ganglion cell layer, the inner nuclear layer, and the two plexiform layers. In addition, regions corresponding to the inner and/or outer segments of the photoreceptor cells showed positive staining for cytokine signaling components. In contrast, the ERK2 protein kinase was found throughout the neonatal retina. In the mature retina, ERK2 was concentrated in the ganglion cells and the inner plexiform layer, while a lesser expression of ERK2 was detected in the inner nuclear layer, the outer plexiform layers, and the outer nuclear layer. CONCLUSIONS: In the neonatal mouse retina, signaling components of the Jak-STAT pathway and ERK2 are differentially expressed. All cytokine signaling components included in this study are expressed in the differentiated inner retina as well as in cells occupying the ventricular zone, suggesting that both postmitotic neurons and proliferative progenitors may directly respond to CNTF-like cytokines during postnatal development. The distribution of cytokine signaling pathway components in the adult mouse retina is consistent with previous findings that ganglion cells and Müller glia are the primary target cell types for CNTF.

Animals↗
PubMed2004

Myristoyl-CoA:protein N-myristoyltransferases: isoform identification and gene expression in retina.

PURPOSE: The mammalian myristoyl-CoA:protein N-myristoyltransferase (NMT) family consists of Type I and Type II enzymes that typically catalyze the addition of myristic acid (14:0) to the N-terminus of specific proteins using myristoyl-Coenzyme A as a donor. However, the N-terminus of certain proteins in frog and bovine retina are modified with fatty acids other than myristic acid, and this utilization of alternative acyl-CoAs has not been found in any other tissues. This alternative use of acyl groups is known as retina heterogeneous acylation and occurs by an unknown mechanism. Therefore, the focus of this work has been to identify NMT isoforms that may be unique or differentially expressed in bovine retina that may utilize acyl-CoAs in addition to myristoyl-CoA. METHODS: We used cDNA library screening, RT-PCR, and PCR to identify NMTs in bovine retina and liver, then compared these proteins to previously described NMTs by amino acid alignments. We used northern blotting to determine the level of Type I and Type II NMTs in a human multi-tissue blot, in bovine retina, and in bovine liver. Southern blotting was done to determine if one or more genomic copies of Type II NMT are present in the bovine genome. A phylogenetic analysis of NMT isoforms is provided to describe the lineage of NMTs from a variety of species. RESULTS: We identified a Type I NMT in retina that is nearly identical to previously described Type I enzymes, as well as a novel Type II enzyme. We further characterized the retina specificity of the novel enzyme by evaluating which Type II enzymes are present in bovine liver. We find that cow liver contains a Type II NMT identical to the previously described NMT found in human liver, as well as a partial clone that is identical to the novel Type II NMT from retina. NMT Type I and II message expression appears to vary in a panel of tissues, and we suggest that retina expresses at least three NMT isoforms, with the Type I short form and Type II being more abundant than the Type I long form NMT. We also present evidence that there are two copies of Type II NMT in the bovine genome. CONCLUSIONS: Our efforts to find a retina-specific NMT resulted in the identification of a previously unknown Type II NMT, but this enzyme also appears to be expressed in liver. Therefore, we are unable to conclude that heterogeneous acylation occurs by means of an exclusive enzyme capable of efficiently utilizing multiple acyl-CoA substrates solely in retina. Nonetheless, our results lead to an improved understanding of protein acylating enzymes and encourage us to further identify factors that influence tissue-specific expression of each isoform and regulate their enzymatic activity in terms of acyl-CoA selectivity and heterogeneous protein acylation.

Acetylation↗
PubMed1975

Immunochemical analysis of water-soluble antigens of chick retina in the course of embryogenesis.

Water-soluble antigens of chick retina were investigated using rabbit antisera to total extract and to individual electrophoretic fractions of retinal extract by methods of immunoelectrophoresis and Ouchterlony precipitation test. In the retina of the adult chick six serum and eleven tissue antigens were demonstrated. The tissue antigens of the retina comprised one organ-specific antigen and ten inter-organ antigens which were characterized by nonuniform distribution in tissues and organs of adult chick. Three antigens out of these were found only in tissues of the eye (retina, iris) and in the brain--inter-organ antigens of 'narrow' specificity. The ohter seven inter-organ antigens were found in tissues of brain and eye, as well as in various tissues and organs of hens--inter-organ antigens of 'broad' specificity. A high degree of antigen similarity between retina and iris was observed. Anti-retina sera in chick lens could detect only inter-organ antigens of 'broad' specificity. In the course of embryogenesis the first to appear in the developing retina were inter-organ antigens of 'broad' specificity (on 3rd day of incubation). Formation of antigens of this group was completed by the 9th day of incubation. On the contrary, inter-organ antigens of 'narrow' specificity appeared later, in the period of histogenesis of retina (from 5 to 18 days of incubation). The organ-specific antigen of retina was found by 7th day of incubation. One of the inter-organ retinal antigens of'narrow' specificity (retina-iris-brain) appeared in the developing chick brain at the same time as in retina--on 10th-11th day of incubation. Using the indirect immunofluorescence antibody technique this antigen was identified in the cytoplasm of retinal cells and brain neurones, but was not detected in the nerve fibres.

Animals↗
PubMed1995

Cone photoreceptor regeneration in adult fish retina: phenotypic determination and mosaic pattern formation.

The retina of anamniotes (fish and amphibia), unlike the CNS of most vertebrates, can regenerate neurons following injury. Using the highly ordered mosaic of single and double cones in the retina of the adult green sunfish (Lepomis cyanellus) as our model system, we examined the events that followed the surgical excision of a small patch of central retina. After surgery there was a transient elevation in the number, and a change in the distribution, of proliferative cells within the retina. The wound was filled in two ways: a proliferative regeneration of new retina and a nonproliferative movement of the wound boundaries toward the center of the lesion. The nonproliferative movement stretched the surrounding, intact retina. In stretched retina the basic pattern of the cone mosaic was maintained, but it was augmented by new cones, even though cones are not normally generated in intact central retina. The stretch itself likely triggered the anomalous cone production. The new and preexisting cones in stretched retina had their morphological phenotypes influenced by mutual contact, often resulting in atypical morphologies (triple and quadruple cones). In the center of the lesioned area, the regenerated cone mosaic was disordered, had a higher than normal cone density, and contained atypical morphologies. The presence of outer segments and synaptic pedicles suggested that the new cones in regenerated and stretched retina were functional. We interpret these results to mean (1) a stretch-induced decrease in cell density can trigger a compensatory, adaptive neurogenesis, (2) cone morphological phenotypes in fish retina are plastic throughout life, and are influenced by cone-cone contacts, (3) the mechanisms that spatially regulate cone production during normal growth are disrupted regeneration.

Animals↗
PubMed1996

Microglial cells invade the outer retina as photoreceptors degenerate in Royal College of Surgeons rats.

PURPOSE: Photoreceptor degeneration is accompanied by the invasion of phagocytic cells into the outer retina of Royal College of Surgeons (RCS) rats. Previous studies suggested that these mononuclear phagocytes were blood-borne macrophages and not retinal pigment epithelial cells nor Müller glia. Thus, immunospecific markers were used to identify these cells and to determine their distribution in the dystrophic retina. METHODS: Retinas from RCS and control (RCS-rdy+) rats were processed for immunocytochemistry using antibodies against phosphotyrosine, which labels both microglial cells and peripheral macrophages, and ED2, which labels peripheral macrophages only. As a positive control to demonstrate ED2-labeling of peripheral macrophages that enter the retina during injury, experiments were performed using needle-punctured Long Evans rat retinas. RESULTS: In normal animals, process-bearing, phosphotyrosine-reactive cells were restricted to the inner retinal layers and the outer plexiform layer. In early dystrophic retinas, phosphotyrosine-reactive cells also were observed in the outer retinal layers. The number of phosphotyrosine-labeled cells in the outer retina increased substantially in later stages of dystrophy. ED2-reactive cells were observed in normal or dystrophic retinas only after needle puncture. CONCLUSIONS: These findings suggest that phagocytic cells during the early stages of dystrophy in RCS rat retinas are derived from resident microglial cells, not from peripheral macrophages. The migration of microglial cells into the outer retina when photoreceptor cells begin to degenerate further suggests that they may play a major role in photoreceptor cell death in the dystrophic retina.

Animals↗
PubMed2000

Somatostatin and somatostatin subtype 2A expression in the mammalian retina.

This review discusses the expression and cellular localization of the neuropeptide somatostatin (SRIF) and one of the SRIF subtype (sst) receptors, sst(2A) in the mammalian retina. SRIF immunoreactivity is predominantly localized to a sparse population of amacrine and displaced amacrine cells in the ganglion cell layer in several mammalian retinas including the rat, rabbit, cat, and primate. These cells, characterized by multiple processes, form a sparse network in the inner plexiform layer (IPL) in all retinal regions. Very few processes are also in the outer plexiform layer. In contrast to the predominant distribution of SRIF processes to the IPL, there is a widespread distribution of sst(2A) immunoreactivity to both the inner and outer retina in all mammalian retinas studied to date. In rabbit retina, sst(2A) immunoreactivity is predominant in rod bipolar cells and in sparse wide-field amacrine cells. In the rat retina, sst(2A) immunoreactivity is localized to several neuronal cell types-cone photoreceptors, horizontal cells, rod and cone bipolar cells, and amacrine cells. Reverse-transcriptase-polymerase chain reaction analysis found that sst(2A) mRNA is expressed in the rat retina, while sst(2B) mRNA is not detected. Finally, in the primate retina sst(2) immunoreactivity is predominant in cone photoreceptors, with additional immunostained cell bodies and processes in the inner retina. These findings indicate that SRIF may modulate several neuronal cell types in the retina, and that it has a broad influence on both scotopic and photopic visual pathways.

Animals↗
PubMed1993

5A11 antigen is a cell recognition molecule which is involved in neuronal-glial interactions in avian neural retina.

In continuing efforts to identify cell-surface molecules involved in cell-cell interactions in the developing avian retina, we identified a monoclonal antibody, the 5A11 antibody, which possessed the ability to interfere with contact-dependent glial cell maturation in vitro. We sought to determine the molecular and biochemical identity of the glycoprotein recognized by this antibody, and using additional criteria, establish whether the 5A11 antigen is indeed a cell-recognition molecule in the developing retina. Immunohistochemical analyses demonstrate that in the hatchling chick retina and in live cultures of embryonic retina cells, the 5A11 antigen is predominantly associated with Müller glial cells whereas little is observed on neuronal elements. Microsequencing of the major component isolated by immunoaffinity chromatography identifies the HT7 antigen (Seulberger et al.: EMBO Journal 9:2151-2158, 1990), a unique member of the immunoglobulin super gene family (IGSF), as a homologous if not identical protein to the 5A11 antigen. The HT7 antibody, furthermore, recognizes affinity purified 5A11 antigen, and both the HT7 antibody and additional probes generated against the 5A11 antigen recognize a major polypeptide of 45.5 kDa and a minor band of 69 kDa on Western blots of membrane preparations from neural retina. To verify that the 5A11 antigen mediates cell-cell recognition events in the developing neural retina, we examined the consequences of adding antibody to monolayer cultures of dissociated embryonic retina cells and to dissociated retina cells in rotation-mediated suspension culture. Addition of the 5A11 antibody to monolayer cultures results in alteration in the development of the stereotypic arrangement of neurons and glia characterized by a reduction in the number and complexity of neural extensions upon the glial-derived flat cells. Similarly, addition of antibodies generated against the 5A11 antigen to dissociated cells in rotation cultures significantly reduces retina cell reaggregation as monitored by computer-assisted image analysis of cell aggregate size. These data and the identification of the 5A11 antigen as a member of the IGSF establish a role for the 5A11 antigen as a novel recognition molecule in the developing neural retina.

Amino Acid Sequence↗