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Light enhances the turnover of phosphatidylinositol in rat retinas.

Light stimulation of isolated rat retinas is shown to enhance the turnover of phosphatidylinositol (PI) as demonstrated by a light-dependent increase in [3H]inositol incorporation and concurrent hydrolysis of existing PI. Studies with rat retinas incubated with [3H]inositol and then microdissected at the level of the outer plexiform layer into photoreceptor cell and inner retina layers indicated that the light-enhanced incorporation of [3H]inositol was associated with the photoreceptor cell layer. The rate of PI hydrolysis in retinas prelabeled in vivo with [3H]inositol was higher in light than in dark incubations and was higher in the photoreceptor cell layer than within the inner retina. Within the photoreceptor cell layer. PI turnover involved 2%/min of the total PI content in dark and 6-8%/min in light. In contrast to what has been reported for stimulus-enhanced turnover of PI in some tissues, this light-enhanced turnover of PI in the retina was not associated with detectable reductions in PI content. Parallel studies of sodium (22Na) uptake demonstrated that the photoreceptor cells remained functional during these incubations as they retained the capacity to restrict the entry of 22Na in light but not in dark.

Animals↗

Muscarinic receptor subclasses in the chick embryo retina: influence of corticosterone treatment.

The present study was performed on retinas of chick embryos receiving at day 8 of incubation an intracerebral injection of 0.02 microgram of corticosterone. We had previously shown with the use of [3H]quinuclidinylbenzilate [( 3H]QNB) that such treatment induced the appearance of two muscarinic binding sites in the treated retinas, whereas only one was detectable in the controls. In the present study we investigated muscarinic cholinergic receptor subclasses with agonist and antagonist binding. Agonist binding was studied by varying the concentrations of carbachol and acetylcholine (10(-9) M-10(-5) M) in the presence of a constant concentration (0.2 nM) of [3H]QNB. Two subpopulations of receptors were revealed, a high- and a low-affinity receptor, in both treated and control retinas. However, in the hormone-treated retinas, the two subpopulations significantly differed from the controls in their affinity and in their relative percentage among the total receptor population. Moreover, using pirenzepine, an antagonist known to have the capacity to distinguish between muscarinic cholinergic subclasses, two receptor subpopulations were found to be present in the hormone-treated retinas but a single one in the controls. It is suggested that hormone treatment can either induce the appearance of a new subclass of muscarinic cholinergic receptors or favor the maturation of a population of retinal cells having these receptors. Pirenzepine binding in retinas from intact embryos of 7, 9, and 11 days of incubation revealed one receptor subpopulation. Thus, these findings are more consistent with the hypothesis that corticosterone effects the target cells, either inducing changes in muscarinic receptor and/or modifying the receptor environment.

Acetylcholine↗

Induced release of gamma-aminobutyric acid by a carrier-mediated, high-affinity uptake of L-glutamate in cultured chick retina cells.

[3H]gamma-aminobutyric acid (GABA) was taken up by cultured embryonic retina cells during the initial stages of cell differentiation. The accumulated GABA was released in the bathing medium and a transient increase in the efflux of GABA was observed when cultures were pulse-stimulated (2 min) with 0.1 mM L-glutamate but not with D-glutamate. The EC50 for L-glutamate to evoke [3H]GABA release was approximately 15 microM. This value is close to the Km for high-affinity uptake of L-glutamate by retina cells. When Na+ ions were replaced by Li+ ions, L-glutamate-induced release of GABA was abolished. Moreover, L-[14C]glutamate uptake by retina cells was significantly reduced when NaCl was replaced by LiCl in the incubation medium. L-Glutamate elicited release of GABA was Ca2+ independent, and was observed when Ca2+ was replaced by Co2+ or when Mg2+ ions were increased to 10 mM concentration. D-Aspartate, which is taken up by the same high-affinity uptake mechanism as L-glutamate, induced an increase in [3H]GABA efflux comparable to L-glutamate. The addition of unlabeled GABA to the medium also promoted the release of accumulated [3H]GABA. However, GABA was twofold less effective than L-glutamate in eliciting [3H]GABA release. The addition of both GABA and L-glutamate to the incubation medium indicated that [3H]GABA efflux due to L-glutamate and GABA was additive. L-Aspartate also promoted an increase in the efflux of [3H]GABA accumulated by retina cells. However, L-aspartate effect was significantly decreased in the absence of Ca2+ or when Na+ ions were replaced by Li+. Our results indicate that at least three releasable pools of GABA are present in the chick embryo retina cells: (a) a GABA-promoted GABA release-homoexchange, (b) a Ca2+-dependent L-aspartate-promoted release, and (c) a Ca2+-independent, Na+-dependent L-glutamate-evoked release. In addition, our data strongly suggest that the L-glutamate-promoted GABA release is due to a process of exchange of L-glutamate with GABA, which may play a fundamental role in the fine control of the excitability of local circuits in the retina.

Animals↗

gamma-Aminobutyric acid system in developing and degenerating mouse retina.

Freeze-dried sections (14 microns thick) of retinal layers were prepared from mice with retinal degeneration (C3H strain) and control mice (C57BL strain). The weighed sections (2-30 ng dry weight) were analyzed using our microassay methods. In the control retina, gamma-aminobutyric acid (GABA) concentration and glutamate decarboxylase (GAD) activity, on a dry weight basis, increased from birth to 9 weeks of age and decreased slightly at 20 weeks. In the degenerated retina, the levels of GABA and GAD activity were higher at birth than in the control retina, and continued to increase until 20 weeks of age, at which time the GAD activity reached a markedly high level. This increase was found when the total GABA and GAD levels per retina were determined. In the normal retinal layers, GABA and GAD were confined primarily to the inner plexiform layer. In the degenerated retina, GAD activity gradually increased in the inner layers during postnatal development, but by 20 weeks the increase was most prominent in the inner part of inner nuclear layer and in the outer part of inner plexiform layer. GABA transaminase activity and its distribution were not much different in both normal and degenerated retinas during development.

Animals↗

Conservation of docosahexaenoic acid in rod outer segments of rat retina during n-3 and n-6 fatty acid deficiency.

We investigated the mechanism by which rat retina conserves docosahexaenoic acid during essential fatty acid deficiency. Weanling female albino rats were fed diets containing either 10% by weight hydrogenated coconut oil, safflower oil, or linseed oil for 15 weeks. Plasma and rod outer segment (ROS) membranes were prepared for fatty acid and phospholipid molecular species analysis. In addition, retinas were removed for morphometric analysis. We found the following: (1) Plasma phospholipids and cholesterol esters from coconut oil, safflower oil, and linseed oil diet groups were enriched in 20:3(n-9), 20:4(n-6), and 20:5(n-3), respectively. The levels of these 20-carbon fatty acids in the ROS, however, were only slightly affected by diet. (2) The fatty acids and molecular species of ROS phospholipids from the safflower oil and coconut oil groups showed a selective replacement of 22:6(n-3) with 22:5(n-6), as evidenced by a reduction of the 22:6(n-3)-22:6(n-3) molecular species and an increase in the 22:5(n-6)-22:6(n-3) species. (3) The renewal rate of ROS integral proteins, determined by autoradiography, was 10% per day for each diet group. (4) Morphometric analysis of retinas showed no differences in the outer nuclear layer area or in ROS length between the three groups. We conclude that the conservation of 22:6(n-3) in ROS is not accomplished through reductions in the rate of membrane turnover, the total amount of ROS membranes, or in the number of rod cells. The retina may conserve 22:6(n-3) through recycling within the retina or between the retina and the pigment epithelium, or through the selective uptake of 22-carbon polyunsaturated fatty acids from the circulation.

Animals↗

Differential glycosylation of the 5A11/HT7 antigen by neural retina and epithelial tissues in the chicken.

The 5A11/HT7 antigen, a member of the immunoglobulin supergene family, has been implicated in heterotypic cell-cell interactions during retina development. Immunopurified 5A11 antigen isolated from Nonidet P-40-solubilized retina membranes had two components as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), a 45.5-kDa doublet and a 69-kDa polypeptide. Immunoreactive bands of 46-50 kDa were recognized following SDS-PAGE of detergent-solubilized membrane proteins from liver, kidney, and erythrocytes. Treatment with N-glycosidase F (EC 3.2.2.18) converted the 45.5-50-kDa immunoreactive polypeptides from all tissues to 32 kDa, indicating that the observed differences in molecular mass were due to differences in glycosylation. N-Glycosidase F treatment also converted the 69-kDa form from retina to 46 kDa, indicating a different polypeptide core than the 32-kDa species. Treatment with endo-beta-N-acetylglucosaminidase H (EC 3.2.1.96) resulted in modest increases in electrophoretic mobility due to hydrolysis of high mannose or hybrid oligosaccharides and lack of hydrolysis of complex oligosaccharides resistant to endo-beta-N-acetylglucosaminidase H digestion. Immunoreactivity was retained after deglycosylation. Much of the difference in molecular weight could be attributed to variations in sialylation. The higher molecular mass species of the 45.5-kDa doublet from retina and the polypeptides from other tissues were susceptible to neuraminidase (EC 3.2.1.18) and O-glycosidase (endo-alpha-N-acetylgalactosaminidase; EC 3.2.1.97) digestion. Labeling with elderberry bark lectin (specific for alpha 2,6-linked sialic acid) was confined to the higher molecular mass species of the 45.5-kDa doublet and was considerably greater in antigen derived from epithelia rather than neural retina. In paraffin sections of chick retina, elderberry bark lectin staining was confined to the retinal pigmented epithelium, photoreceptor cells, and bipolar cells with no staining of the Müller cells, which bear the bulk of the 5A11 antigen. These results indicate tissue-specific posttranslational modifications, particularly differences in sialylation of antigen-bearing polypeptides.

Animals↗

The uptake of 3Hp -aminobutyric acid by the retina.

1. The accumulation of (3)H-gamma-aminobutyric acid (GABA) by the isolated rat retina has been measured.2. When retinae were incubated at 37 degrees C in a medium containing (3)H-GABA, tissue:medium ratios of about 25:1 were attained after a 30 min incubation.3. After incubations of 40 min at 37 degrees C, almost all (98%) the radioactivity in the tissue was present as unchanged (3)H-GABA.4. The process responsible for (3)H-GABA uptake showed many of the properties of an active uptake system: it was temperature-sensitive, required the presence of sodium ions in the external medium, was inhibited by anoxia, dinitrophenol and ouabain, and showed saturation kinetics.5. The estimated Km value of GABA was 4.0 x 10(-5)M, and V(max) was 0.167 (mumoles/min)/g retina.6. The uptake of (3)H-GABA was not affected by the presence of large molar excesses of glycine, L-glutamate, L-aspartate, L-alanine, L-proline, or L-histidine, but was inhibited by DL-gamma-amino-beta-hydroxybutyrate, beta-guanidinopropionate, and L-2,4-diaminobutyrate.7. The retina was capable of achieving a large net uptake of GABA, indicating that the accumulation of (3)H-GABA by the tissue was not due only to an exchange process with the endogenous GABA pool.8. The uptake of (3)H-GABA occurred only in tissue from the central nervous system. Thus, retina and cerebral cortex rapidly accumulated radioactivity, but slices of cornea, posterior wall of the eye, and liver achieved tissue: medium ratios of approximately one.9. There was a rapid efflux of radioactivity from retinae placed in fresh medium and after 60 min, 90% of the radioactivity was lost from the tissue. The radioactivity released into the medium was present largely as (3)H-acidic and neutral metabolites. When the metabolism of GABA was inhibited by the presence of amino-oxyacetic acid in the medium, only about 10% of the radio-activity was lost from the tissue during a similar 60 min incubation, and the radioactivity released was present largely as unchanged (3)H-GABA.10. It is suggested that the GABA uptake process may represent a possible mechanism for the inactivation of GABA if this amino acid is released at inhibitory synapses in the retina.

Acetates↗

Substance P in the bovine retina: localization, identification, release, uptake and receptor analysis.

Substance P-like immunoreactivity is restricted to at least three different cell types in the bovine retina. They are amacrine cells, displaced amacrine cells and interplexiform neurones. High performance liquid chromatography and radioimmunoassay have shown that the bovine retina substance P-like immunoreactive material cannot be distinguished from authentic substance P. Isolated bovine retina does not take up exogenous [3H]substance P as revealed by autoradiography. By increasing the external K+ concentration it was shown that the release of endogenous substance P from the retina was stimulated. This release is Ca2+ dependent. The binding of [3H]substance P to crude membrane preparations of bovine retina was saturable and revealed a single population of binding sites with a KD value of 0.32 nM and a Bmax of 2.45 pmol/g wet wt. The characteristics of the binding site suggest the presence of substance P receptors. It is suggested that substance P is a likely neurotransmitter in the bovine retina.

Animals↗

Regional variation of contrast sensitivity across the retina of the achromat: sensitivity of human rod vision.

1. Detection thresholds for two-dimensional Gabor functions of varying spatial and temporal frequency were used to investigate the post-receptoral sensitivity across the retina of the typical and complete achromat. 2. Under photopic conditions there is no evidence for post-receptoral cone function at any retinal eccentricity investigated. Sensitivity saturates in a way consistent with known psychophysical and electrophysiological measures of rod saturation. This occurs in a unitary fashion across the retina. 3. Under scotopic conditions the regional fall-off in spatio-temporal sensitivity is similar for the achromat and duplex retina. This suggests that the rods in the achromat make normal neural connections. 4. Taken together this supports the contention that the typical and complete achromat is a functional rod monochromat and hence can be used to explore the sensitivity of the isolated rod post-receptoral mechanism under mesopic conditions where its sensitivity is optimal. This is where its contribution is most difficult to isolate in the duplex retina. 5. For the human rod mechanism, mesopic post-receptoral sensitivity for all spatio-temporal stimuli is optimal in the central region of the retina and falls off as a function of eccentricity. 6. For localized stimuli, peripheral spatial sensitivity is reduced evenly at all spatial frequencies compared with that of the central retina. A similar displacement of the spatial sensitivity function of the rod mechanism occurs as illuminance is reduced. 7. For localized stimuli, temporal acuity of the rod mechanism is around 20-25 Hz irrespective of retinal position. As the illuminance is further lowered dynamics of the rod pathway are reduced irrespective of retinal position and the sensitivity function maintains a bandpass shape. 8. The regional distribution sensitivity of the rod mechanism changes as illuminance is reduced from mesopic to scotopic levels.

Color Vision Defects↗

Developmental control of transcription of a retina-specific gene, QR1, during differentiation: involvement of factors from the POU family.

Developmental control of gene expression often results from the coupling of growth arrest with the establishment of differentiation programs. QR1 is a gene specifically expressed in retinas during the late phase of embryogenesis. At this stage neuroectodermal precursors have reached terminal mitosis and are undergoing differentiation into distinct cell types. Transcription of the QR1 gene is tightly regulated during retinal development: this gene is expressed between embryonic day 9 (ED9) and ED17 and is completely repressed at hatching in quail. Moreover, QR1 transcription is downregulated when postmitotic neural retina cells are induced to proliferate by pp60v-src. We studied the stage-dependent transcriptional control of this gene during quail neural retina (QNR) cell development. Transient transfection experiments with QR1/CAT constructs at various stages of development showed that a region located between -935 and -1265 bp upstream of the transcription start site is necessary to promote transcription in retina cells during the late phase of embryonal development (QNR9, corresponding to ED9). By in vivo footprinting assays we identified at least two elements that are occupied by DNA-protein complexes in QNR cells: the A and B boxes. The A box allows formation of several biochemically distinct complexes: C1, C2, C3, and C4. Formation of the C2 complex mainly during early stages (ED7) and of C2, C3, and C4 complexes during postnatal life correlates with repression of QR1 transcription, whereas the C1 complex is strongly induced at ED11 when the QR1 gene is expressed. We previously showed that C1 was involved in downregulation of QR1 transcription by pp60v-src. Several complexes are also formed on the B box. We show that these complexes are exclusively present in neural tissues and that they involve members of the POU family of transcription factors. Mutations of each one of the two regions which abolish the binding of the C1 factor(s) on the A box and of the POU factor(s) on the B box also prevent stimulation of QR1 transcription in QNR9. Therefore, both elements appear to be required for the stage-specific transcription of the QR1 gene. We also show that the regulatory region from position -1265 to position -935 is able to confer stage-specific transcription upon a heterologous promoter (thymidine kinase). Indeed, this region stimulates transcription in differentiating retinas (QNR9) and represses transcription in terminally differentiated retinas (QNR17, corresponding to postnatal life). Our results suggest that cell growth regulation and developmental control are coordinated through the A and B boxes in regulating QR1 transcription during retinal differentiation.

Animals↗

Growth factor staining patterns in the pig retina following retinal laser photocoagulation.

AIM: To identify changes in growth factor expression in miniature pig retinas following retinal laser photocoagulation. METHODS: Pigs were sacrificed at different times (15 minutes to 42 days) post-laser and the retinas were immunolabelled for basic fibroblast growth factor, insulin-like growth factor I, transforming growth factor beta, epidermal growth factor, transforming growth factor alpha, platelet derived growth factor, vascular endothelial growth factor, and epidermal growth factor receptor. Total mRNA levels were also determined. RESULTS: With the exception of vascular endothelial growth factor, immunoreactivity for all other growth factors studied and epidermal growth factor receptor was observed throughout normal non-lasered control retina, generally being high in the retinal pigment epithelium and low in the neural retina. Changes in growth factor expression following laser photocoagulation were observed only in burn areas and changes were mainly confined to the retinal pigment epithelium and outer nuclear layer. The immunoreactivity within retinal pigment epithelial cells in burn areas was either absent or decreased following laser treatment but returned to normal by 21 days. The immunoreactivity was increased within the outer nuclear layer of burn areas during the healing process but returned to normal by 42 days. Vascular endothelial growth factor immunoreactivity was weak/absent in the normal retina and remained unchanged following laser photocoagulation. Change of total mRNA levels in burn areas during time post-laser was confined to retinal pigment epithelial cells, being low immediately following photocoagulation and returning to normal by 42 days. CONCLUSIONS: These results demonstrate a temporal alteration in growth factor expression and transcriptional activity in the retina following laser photocoagulation.

Animals↗

New neuroretinal rim blood flow evaluation method combining Heidelberg retina flowmetry and tomography.

AIM: Accurate Heidelberg retina flowmeter (HRF) measurements require correct manual setting of the HRF photodetector sensitivity. The neuroretinal rim produces a weak signal relative to the peripapillary retina. A newly developed HRF alignment and sensitivity protocol, capable of accurate rim measurement, was investigated. METHODS: 18 eyes of nine healthy volunteers were examined by HRF. Three images of each eye were taken using three different imaging methods. Method 1: a conventional image (optic nerve head centred image with photodetector sensitivity optimised for the strong signal from the peripapillary retina); method 2: the setting of method 1 with photodetector sensitivity optimised for the weak signal from the rim; and method 3: the setting of method 2 with the temporal rim margin tangent to the lateral image border to remove the overpowering signal from the temporal peripapillary retina. The neuroretinal rim was defined by the Heidelberg retina tomograph (HRT). Blood flow and reflectivity values (DC component) in the rim area were compared for the three methods using pointwise analysis. Coefficients of variation of repeated measurements in 12 subjects have been calculated for method 3. RESULTS: The neuroretinal rim area measured by method 1 had a significantly lower brightness compared with method 2 and 3 (p=0.0002 and p=0.0002, respectively). Method 2 provided proper sensitivity for the weak signals of the rim area based on rim tissue DC values; however, this sensitivity setting was too high for the strong signal from the peripapillary retina. Method 3 avoided the strong peripapillary signal with the proper signal from the rim and provided significantly higher flow values of the rim area at 75 and 90 percentile pixels (p=0.0065 and p=0.0038 respectively) compared with method 2. Interobserver repeatability ranged from 16.85% to 21.96% for the different parameters. CONCLUSIONS: Method 3 provides an accurate and reproducible flow measurement of the neuroretinal rim area through proper sensitivity for the weak rim signal, alignment, and removal of the strong temporal signal from the image. This new method is recommended to improve accuracy of blood flow measurement in the neuroretinal rim.

Adolescent↗

Scotopic threshold response of proximal retina in cat.

We describe a new response in proximal retina of cat that is present under scotopic conditions, clearly differs from PII (b-wave and DC component) and contributes a negative potential at threshold to the diffuse electroretinogram (ERG). We have termed this response the scotopic threshold response (STR). Extracellular potentials evoked in response to circular spots of light at dark-adapted threshold, and with dim backgrounds, were recorded with microelectrodes placed intraretinally at different depths. The dark-adapted response of proximal retina (STR) consisted of a graded negative potential to the onset of illumination that maintained amplitude during illumination and decayed back toward the base line at stimulus offset without evidence of a negative-going off response. It thereby differed in form from the photopic M-wave response of proximal retina, which has a negative-going off response. It also did not exhibit spatial tuning, simply increasing in size with stimulus area. In addition, the STR appears to be a rod-driven response whose threshold approximates that of the most sensitive ganglion cells in cat, whereas the M-wave is a much higher threshold cone-driven response. The STR could be clearly distinguished from PII on the basis of its form, depth-distribution, and dynamic range. For example, the STR had its maximum amplitude in the proximal retina at 17% retinal depth, whereas scotopic PII had its maximum in the distal retina at 48% retinal depth. Also, the STR had a lower threshold than PII intraretinally and saturated well below the level of saturation of scotopic PII (rod saturation). By analogy to the PNR and M-wave, the STR is hypothesized to represent either an extracellular voltage arising from proximal retinal neurons or Müller cell responses to K+ released by these neurons. Recording in the vitreous, near the retinal surface, showed that the STR always had a negative polarity. The polarity reversal of the STR at 50-60% retinal depth (from negative, proximal to positive, distal) suggested the presence of a sink proximal to the reversal point and a source distal to it. We also recorded the vitreal ERG in response to diffuse illumination of the dark-adapted retina. The STR could be clearly identified in the scotopic ERG as a threshold negative potential that had been observed previously in the mammalian ERG. The STR differs, therefore, from PII (b-wave and DC component) that is a higher threshold positive component in the diffuse ERG.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Intraretinal analysis of the threshold dark-adapted ERG of cat retina.

1. The dark-adapted electroretinogram (ERG) obtained with 4-s flashes near threshold and at low intensities has three clear negative components: 1) a fast negative response at stimulus onset, 2) a slow negative response that increases in amplitude for about 2 s, and 3) a negative-going OFF response. The fast negative response was previously shown to originate from the scotopic threshold response (STR) arising in the proximal portion of neural retina. We investigated the intraretinal origins of the other two negative components and found that they also have origins in neural retina proximal to the photoreceptors. 2. In this study the ERG evoked in response to diffuse illumination of the dark-adapted cat retina was recorded between a chlorided silver wire in the vitreous and a plate behind the eye. Extracellular field potentials were recorded simultaneously with a microelectrode placed intraretinally at different retinal depths. In some cases light-dependent changes in extracellular K+ concentration ([K+]o) were recorded with the K+-selective barrel of a double-barreled microelectrode. 3. The three negative-going ERG potentials increased in amplitude from threshold up to approximately 6.0 log q.deg-2.s-1. The potential during illumination then became nearly flat, and the negative OFF response was exaggerated. At higher intensities, positive-going PII (DC-component and b-wave) and the c-wave emerged, whereas the a-wave (negative-going) was usually below threshold. 4. The slow negative ERG response was generated in neural retina, rather than by the retinal pigment epithelium. The response was positive-going in the transretinal recordings obtained in subretinal space. It was not, however, related to two positive-going responses from distal retina previously described as contributing negative components to the ERG: the neural retina component of the c-wave, termed slow PIII, and the rod-receptor potential. The slow negative response had a slower time course and lower threshold than slow PIII, and, more importantly, it was present in the absence of the [K+]o decrease in subretinal space that causes slow PIII. Furthermore, APB (1.2 mM vitreal concentration) blocked the entire negative-going ERG, whereas at higher intensities it blocked PII but not the c-wave in the ERG or slow PIII in transretinal recordings. These results indicate that the negative-going ERG components near threshold and at low intensities all originated proximal to the photoreceptors.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

First appearance of orthogonal arrays of particles in Müller cell membranes depends on retina maturation.

A freeze-fracture analysis was undertaken to investigate both the appearance of orthogonal arrays of particles (OAP) in the Müller cell endfoot membrane at the retina-vitreous border and the endfoot size during development of two avian species. Chickens and pigeons have a common length of development until hatching (21 days), but differ in maturation velocity. In the chicken, the first OAP appear in the central retina at embryonic day 18 (E18), in the peripheral retina at E19. The OAP density increases rapidly. In the pigeon, the first OAP appear 6 days after hatching when the eyes are opened. In both species, the size of endfeet is maximal when the OAP first appear and is reduced thereafter. In the chicken, during the time up to adulthood both the mean endfoot size and the retina area increase 6-fold suggesting that the number of endfeet remains constant during growth of their individual area. In the pigeon, after an initial increase of mean endfoot area this decreases until adulthood, despite continuous growth of the eye. This strongly suggests that during retina growth the number of Müller cell endfeet increases. It is concluded that these alterations of Müller cell membranes reflect some functional aspects of retina maturation.

Animals↗

Age-dependent differences in cognin regeneration on embryonic retina cells: immunolabeling and SEM studies.

The retina cognin (a glycoprotein isolated from the surface membrane of neural retina cells of chick embryos and postulated to mediate self-recognition and histogenetic association of retina cells) has been visualized by SEM on the surface of embryonic retina cells in vitro following immunolabeling of the cells with antibodies to the purified cognin and with polystyrene latex microbeads. Trypsin dissociation of retina tissue into separated cells resulted in cognin depletion from the cell surface; following incubation at 37 degrees C the cells regenerated the cognin. Regeneration was fastest and most abundant on cells from the youngest retinas examined; it declined markedly with the embryonic age of the cells, suggesting an age-dependent decrease in cell capacity for cognin formation. Evidence is discussed that the rate and amount of cognin regeneration on the cell surface are temporally-causally correlated with the capacity of the cells to reaggregate into retinotypic tissue. The results support the suggested role of the cognin in the mechanism of self-affinity and morphogenetic association of embryonic neural retinal cells.

Animals↗

Postnatal maldevelopment of the retina.

The effect of certain factors on the early postnatal development of the retina was studied by electron microscopy. Although the retina of developing rats injected with sodium iodate on the 1st and 7th postnatal days showed no retardation of development of photoreceptor outer segment, it began to degenerate about 3 weeks after birth. The retina of developing rats exposed to strong light from the 1st to 5th postnatal day revealed slight developmental retardation of the photoreceptor outer segment and the photoreceptor outer segment began to degenerate about 3 weeks after birth. The vitamin A deficient developing rat it began to degenerate nearly 3 weeks after birth. In spite of induction of developmental retardation of the photoreceptor outer segment of the retina, development of the photoreceptor outer segment was not completely inhibited by moderately acting factors. However, 3 weeks after birth, the photoreceptor outer segments began to degenerate. Therefore, some factors during the early postnatal period can cause the late appearance of degeneration of the retina. The characteristics of retinal degeneration are the same as those found in the retinas of adult rats which are affected by each factor. These degenerations later recover morphologically.

Animals↗

Light damage in the rat retina: glial fibrillary acidic protein accumulates in Müller cells in correlation with photoreceptor damage.

Low intensity diffuse white fluorescent light (1,000 lx for 2 h) exclusively induced photoreceptor damage in the inferior retina of albino rats; the temporal retina showed extensive damage, whereas the nasal retina revealed threshold lesions prior to recovery. To expand our morphological data, further experiments were undertaken to determine if glial fibrillary acidic protein (GFAP) expression was associated with the regions of photoreceptor damage. To follow the time course of GFAP expression, immunoblot analysis was carried out on retinal homogenates of dark-adapted (control) rats and light-exposed rats returned to cyclic light for 0 h, 1, 2, 3 and 6 days. A significant twofold increase in GFAP immunoreactivity over controls was observed in the retinas of light-exposed rats returned to cyclic light for 6 days. Using an indirect immunohistochemical method, retinal sections of the control and light-exposed rats allowed to recover for 6 days were stained for GFAP. GFAP immunoreactivity was localised to the astrocytes and Müller cells. Moreover, GFAP staining in Müller cells in the retinas of control animals was uniformly restricted to rare end-feet. In contrast, a gradient of GFAP immunoreactivity was observed in experimental rats, rising from the superior retina to the inferior temporal quadrant; the GFAP staining in the inferior nasal quadrant was intermediate. Thus, GFAP immunoreactivity was proportional to photoreceptor damage. Interestingly, no GFAP induction could be demonstrated in the pineal glands of light-exposed rats.

Animals↗