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The epithelial sodium channel (ENaC) in rodent retina, ontogeny and molecular identity.

PURPOSE: To assess the appearance of the epithelial sodium channels (ENaC) during retinal development and establish its molecular identity. METHODS: Photoreceptors were isolated by horizontal sectioning of rat retina with a vibratome series 1000. Intact retina and isolated photoreceptors were analyzed for the developmental appearance of ENaC using the Polymerase Chain Reaction (PCR) technique. Immunofluorescence was conducted with the aid of an antibody raised against the 14 amino acids peptide QGLGKGDKREEQGL, corresponding to the N-terminal region (44-58 aa) of alpha ENaC. RESULTS: ENaC message was present in rat retina photoreceptors, isolated just one day after birth. The bipolar and ganglion cell layers, separated from whole retina by vibratome, also contained the ENaC message. The partial sequence of the photoreceptor ENaC (496 base) exhibited one hundred percent homology with the channel from rat known sources. Immunochemistry revealed that only the outer nuclear layer was positive for the ENaC in one-day-old rat; the inner segment became immunopositive at the age of 9 days. CONCLUSION: The ENaC is present in the retina and visible soon after birth. These observations suggest that ENaC may prove to have physiological importance in the retina but until now its function is unknown.

Amino Acid Sequence↗

Rhodopsin-mediated blue-light damage to the rat retina: effect of photoreversal of bleaching.

PURPOSE: Acute white-light damage to rods depends on the amount of rhodopsin available for bleaching during light exposure. Bleached rhodopsin is metabolically regenerated through the visual cycle involving the pigment epithelium, or photochemically by deep blue light through photoreversal of bleaching. Because photoreversal is faster than metabolic regeneration of rhodopsin by several orders of magnitude, the photon catch capacity of the retina is significantly augmented during blue-light illumination, which may explain the greater susceptibility of the retina to blue light than to green light. However, blue light can also affect function of several blue-light-absorbing enzymes that may lead to the induction of retinal damage. Therefore, this study was conducted to test whether rhodopsin and its bleaching intermediates play a role in blue-light-induced retinal degeneration. METHODS: Eyes of anesthetized rats and mice that did or did not contain rhodopsin were exposed to green (550 +/- 10 nm) or deep blue (403 +/- 10 nm) light for up to 2 hours. Rats with nearly rhodopsinless retinas were obtained by bleaching rhodopsin in animals with inhibited metabolic rhodopsin regeneration-that is, under halothane anesthesia. In addition, Rpe65(-/-) mice that are completely without rhodopsin were used to test the susceptibility to blue-light damage of a rodent retina completely devoid of the visual pigment. Effects of illumination on photoreceptor morphology were assessed 24 hours or 10 days thereafter by morphologic and biochemical methods. RESULTS: Exposure to blue light resulted in severe retinal damage and activation of the transcription factor AP-1 in rats. In contrast, green light had no effect. When rhodopsin was almost completely bleached by short-term green-light exposure while metabolic regeneration (but not photoreversal) was prevented by halothane anesthesia, blue-light exposure induced distinct lesions in rat retinas. When both metabolic rhodopsin regeneration and photoreversal of bleaching were almost completely inhibited, blue-light exposure caused only very moderate lesions. When mice without rhodopsin were exposed to blue light, no damage occurred, in contrast to wild-type control mice. CONCLUSIONS: Short time exposure to blue light has deleterious effects on retinal morphology. Because damage was observed only in the presence of the visual pigment, blue-light-induced retinal degeneration is rhodopsin mediated. Absorption of blue light by other proteins is not sufficient to induce light damage. Photoreversal of bleaching, which occurs only in blue but not in green light, increases the photon-catch capacity of the retina and may thus account for the difference in the damage potential between blue and green light.

Animals↗

Circadian clock regulation of pH in the rabbit retina.

Although it is generally accepted that the acid-base ratio of tissue, as represented by the pH, is strictly regulated to maintain normal function, recent studies in the mammalian nervous system have shown that neuronal activity can result in significant shifts in pH. In the mammalian retina, many cellular phenomena, including neuronal activity, are regulated by a circadian clock. We thus investigated whether a clock regulates retinal pH, using pH-sensitive microelectrodes to measure the extracellular pH (pH(o)) of the in vitro rabbit retina in the subjective day and night, that is, under conditions of constant darkness. These measurements demonstrated that a circadian clock regulates the pH(o) of the rabbit retina so that the pH(o) is lower at night than in the day. This day/night difference in retinal pH(o) was observed when the rabbits were maintained on a normal light/dark cycle and after they were maintained on a light/dark cycle that was phase-delayed by 9 hr. Continuous recordings of retinal pH(o) around subjective dusk indicated that the change from daytime to nighttime pH(o) is relatively fast and suggested that the clock that regulates pH(o) is located in the retina. The lowest pH(o) recorded in the retina in both the day and night was in the vicinity of the inner segments of photoreceptor cells, supporting the idea that photoreceptors serve as the primary source of protons. The circadian-induced shift in pH(o) was several times greater than light-induced pH(o) changes. These findings suggest that a circadian clock in the mammalian retina regulates retinal pH.

Animals↗

Oxidative stress induces heme oxygenase-1 immunoreactivity in Müller cells of mouse retina in organ culture.

PURPOSE: Heme oxygenase (HO)-1 immunoreactivity (IR) was examined in normal untreated retina and in retinal explants after in vitro treatment with stress agents. METHODS: Enucleated eyes from young adult C3H mice were immediately fixed and cryosectioned and the retina sections processed for immunocytochemistry with antibodies against HO-1 and glial fibrillary acidic protein (GFAP). From other eyes retinas were isolated and maintained in organ culture, either untreated for 4 days maximum or for 21 hours during which the explants were treated the first 3 hours with selected doses of sodium arsenate or hydrogen peroxide. Thereafter, the explants were processed identically with the normal tissue. RESULTS: In the normal retina, HO-1 and GFAP IR was very low. The culturing itself resulted in an increase in both HO-1 and GFAP immunolabeling in Müller cells of explanted retinas. Both sodium arsenate and hydrogen peroxide further induced strong HO-1 IR in Müller cells but not in other retinal cells. In contrast to HO-1, GFAP staining in Müller cells was not altered as a result of treatment, either by sodium arsenate or hydrogen peroxide at any concentration used. CONCLUSIONS: The results show for the first time that HO-1 can be induced in the retina in vitro by conditions of oxidative stress and that enzyme expression is confined exclusively to Müller cells.

Animals↗

Localization of the alpha(1F) calcium channel subunit in the rat retina.

PURPOSE: The molecular identity of the calcium channels that mediate glutamate release from photoreceptors is unknown. Mutations in the recently identified, retina-specific alpha(1F) calcium channel subunit cause incomplete X-linked congenital stationary night blindness (CSNB2), the phenotype of which is consistent with a defect in neurotransmission within the retina. The purpose of this study was to determine the cellular distribution of the alpha(1F) subunit in the retina. METHODS: Antibodies were raised against a unique peptide from the human alpha(1F) sequence. Rat retina sections were labeled with affinity-purified alpha(1F) antibodies and the immunofluorescence analyzed by confocal microscopy. The alpha(1F) staining was compared with that obtained with a pan-alpha(1) antibody, used to reveal the distribution of known voltage-gated calcium channels in the retina. Some sections were double labeled for alpha(1F) and the photoreceptor synaptic ribbon marker, bassoon. RESULTS: Staining of retina sections with anti-alpha(1F) resulted in strong punctate labeling in the outer plexiform layer (OPL) and weak punctate labeling in the inner plexiform layer (IPL), consistent with a synaptic localization. Staining was also observed in the outer nuclear layer. Within the OPL, alpha(1F) immunoreactivity was clustered in discrete, horseshoe-shaped patches, the shape and dimensions of which are characteristic of rod active zones. Similar structures were labeled with the pan-alpha(1) antibody. Localization of alpha(1F) immunoreactivity to rod active zones was confirmed by double labeling for bassoon, a component of photoreceptor synaptic ribbons. CONCLUSIONS: The distribution of alpha(1F) immunoreactivity in the OPL suggests that calcium influx through alpha(1F) or alpha(1F)-like channels mediates glutamate release from rod photoreceptors.

Animals↗

The fate of heterotopically grafted neural precursor cells in the normal and dystrophic adult mouse retina.

PURPOSE: To study the integration and differentiation of heterotopically transplanted neural precursor cells in the retina of adult mouse mutants displaying apoptotic degeneration of photoreceptor cells. METHODS: Neural precursor cells were isolated from the spinal cord of transgenic mouse embryos ubiquitously expressing enhanced green fluorescent protein. Cells were expanded in vitro and transplanted into the retina of adult wild-type and age-matched beta2/beta1 knock-in mice. Beta2/beta1 knock-in mutants display apoptotic death of photoreceptor cells and were generated by placing the cDNA of the beta1 subunit into the gene of the beta2 subunit of Na,K-ATPase. The integration and differentiation of grafted cells in recipient retinas was studied 1 or 6 months after transplantation. RESULTS: Mutant retinas contained more donor-derived cells than wild-type hosts. Moreover, in mutants, donor cells integrated into deeper retinal layers. In both genotypes, grafted cells differentiated into astrocytes and oligodendrocytes. Only a few ganglion cell axons were myelinated by donor-derived oligodendrocytes 1 month after transplantation, whereas extensive myelination of the nerve fiber layer was observed 6 months after transplantation. Unequivocal evidence for differentiation of grafted cells into neurons was not obtained. CONCLUSIONS: Heterotopically transplanted neural precursor cells are capable of integrating, surviving, and differentiating into neural cell types in normal and dystrophic retinas of adult mice. The particular environment of a pathologically altered retina facilitates integration of transplanted precursor cells. In principle, neural precursors may thus be useful to substitute for or replace dysfunctional or degenerated cell types. Results of the present study also indicate that replacement of retinal cell types is likely to require more appropriate donor cells, such as retinal precursor cells.

Actins↗

[Ultrastructure of melanocytes from retina and choroid of the Pacific salmon].

The ultrastructure of the retina and choroid cells in three species of the Pacific salmon, Oncohrynchus gorbuscha, O. keta and O. masou, was studied. The structure of retina pigment cells is similar in all the three species, only a small difference was found in the percentage of lengthened and rounded forms of melanosomes. Melanocytes of the masu salmon differ also in the structure of their nuclei. The pigment cells of choroid differ from those of retina by a more extended form of melanosomes and by the presence of less rounded melanosomes. In the chum salmon retina we found electron lucent "contact vesicles", whose assignment is open to discussion. In retina pigment cells of the masu salmon smolts ready for migration, the activity of Golgi appararus rises, mitochondria and mature melanosomes increase in number. The choroid pigment cells are slightly swollen, their processes more often and deeper penetrate into the walls of vessels, down to the endothelium. Results of the experiment with the application of an artificial magnetic field (AMF) have shown that the retina and choroid pigment cells in the masu salmon fry react to the field of a certain direction. The phenomenon of magnetosensitivity of pigment cells is discussed in addition to their possible involvement in magnetoreception.

Animals↗

Distribution of organic anion-transporting polypeptide 2 (oatp2) and oatp3 in the rat retina.

PURPOSE: To examine the expression of multifunctional Na(+)-independent organic anion-transporting polypeptides, termed oatp1, oatp2, and oatp3, involving the transport of thyroid hormone in the rat retina at the protein and mRNA levels. METHODS: Northern blot analysis was performed using oatp1, -2, and -3 cDNAs. Reverse transcription-polymerase chain reaction (RT-PCR) was also performed using gene-specific primers for oatp1, -2, and -3. mRNA distribution of these oatps in the rat retina was examined by in situ hybridization. Western blot analysis and immunohistochemistry were also performed by raising specific antibodies against oatp2 and -3. RESULTS: Northern blot analysis showed that the mRNAs for oatp2 and -3 were expressed in the rat retina and retinal pigment epithelium (RPE). Amplified cDNA products by RT-PCR for oatp2 and -3 were also detected in the rat retina-RPE. In contrast, no specific band for oatp1 was detected by Northern blot analysis or RT-PCR. By in situ hybridization, oatp2-specific mRNA signals were seen in the RPE and inner nuclear layer, whereas the oatp3 mRNA signal was localized to the ganglion cell. At the protein level, a single band for oatp2 and -3 proteins was detected in the rat retina-RPE by Western blot analysis. Immunohistochemistry revealed that oatp2 immunostaining was predominantly expressed at the apical surface of the RPE. Weak immunostaining for oatp2 was also seen in the inner nuclear layer and the ganglion cell layer. In contrast, apparent immunostaining for oatp3 was seen in the nerve fiber layer, ganglion cell layer, inner plexiform layer, and outer aspect of the inner nuclear layer. In addition, oatp3 immunostaining was detected predominantly in the optic nerve fiber. CONCLUSIONS: These results reveal that oatp2 is localized mainly in the RPE, suggesting a role for organic anion transport in this specialized ocular tissue. In contrast, oatp3 is localized mainly in optic nerve fibers, suggesting that oatp3 is a specific transporter in the visual nervous system. In conclusion, these data suggest that oatp2 and -3 may be involved in the transport of thyroid hormone in the rat retina.

Animals↗

Topical flunarizine reduces IOP and protects the retina against ischemia-excitotoxicity.

PURPOSE: To determine whether topical application of flunarizine reduces intraocular pressure (IOP) and acts as a retinal neuroprotectant and to compare the effectiveness of flunarizine with betaxolol and nifedipine at reducing the influx of calcium and sodium. METHODS: Ischemia was delivered to the rabbit retina by raising the IOP. After 3 days, a flash electroretinogram (ERG) was recorded, and the retina processed for the localization of certain antigens. In the rat, N-methyl-D-aspartate (NMDA) was injected intravitreally, and 8 days later, the retinas were analyzed for the localization of Thy-1 or the relative amounts of mRNAs for antigens located to ganglion cells or photoreceptors. Rats and rabbits received topical flunarizine or vehicle before and after ischemia or NMDA. IOP was measured in rabbits after a single topical application of 2% flunarizine. Studies were conducted on isolated rat retinas, cortical cultures, and brain synaptosomes to compare the effectiveness of flunarizine with nifedipine and betaxolol at reducing the influx of calcium or sodium. RESULTS: Changes in rabbit retinal choline acetyltransferase and parvalbumin immunoreactivities and the b-wave of the ERG caused by ischemia-reperfusion were blunted by topical treatment with flunarizine. Similarly, NMDA induced reductions in Thy-1 immunoreactivity and mRNA for rat ganglion cell antigens (Thy-1 and neurofilament light form) were counteracted by topical application of flunarizine. Topical application of 2% flunarizine significantly lowered the IOP in rabbits over a period of 5 hours. Flunarizine was more effective than betaxolol and much stronger than nifedipine at attenuating veratridine-induced influx of sodium into synaptosomes. Nifedipine, flunarizine, and betaxolol all reduced the NMDA-induced influx of calcium into the isolated retina or cortical neurons, but betaxolol was the least effective. CONCLUSIONS: Topically applied flunarizine reduces IOP and attenuates injury to the whole of the retina, including the ganglion cells. The neuroprotective action of flunarizine is to reduce the influx of calcium and sodium into stressed neurons. The potent effect of flunarizine on sodium influx would be particularly protective to axons.

Administration, Topical↗

[A study on the activation of nuclear factor-kappa B in rat retina induced by interleukin-1 beta].

OBJECTIVE: To evaluate the inductive role of interleukin (IL)-1 beta and the inhibitory role of pyrrolidine dithiocarbamate (PDTC) on the activation of nuclear factor (NF)-kappaB in the rat retina. METHODS: Sixteen Sprague-Dawley (SD) rats were randomly divided into two groups. The inductive role of IL-1 beta was observed in group A, in which 5 microl IL-1 beta (5 x 10(7) U/L) was injected intravitreally in the right (experimental) eye and PSS 5 microl in the left (control) eye. The inhibitory role of PDTC was observed in group B. In this group, 10 micro1 PDTC (1 mmol/L) was injected intravitreally in the right (experimental) eye and PSS 5 microl in the left (control) eye 1 hour before the intraocular injection of 5 microl IL-1 beta (5 x 10(7) U/L) in the bilateral eyes, respectively. Nuclear extracts was prepared from the retina 4 h and 24 h post-IL-1 beta challenge, and electrophoretic mobility shift assay (EMSA) was used to examine NF-kappa B/DNA binding activity in the retina. RESULTS: At 4 hours after IL-1 beta challenge, NF-kappa B/DNA binding activity increased in the retina of the experimental eyes in group A, and decreased in the experimental eyes of group B, compared to that of their control eyes. At 24 hours after IL-1 beta challenge, NF-kappa B/DNA binding activity in the experimental eyes of group A and in the control eyes of group B was lower than that at 4 hours, respectively. CONCLUSIONS: NF-kappa B in the retina can be activated by intraocular IL-1 beta. NF-kappa B may play an important regulative role in the mechanism of inflammatory reaction in the retina.

Animals↗

[An immunohistochemical study of localization of the calcium-binding protein recoverin in retina of the newt Pleurodeles waltl].

The presence and localization of the calcium-binding protein recoverin, initially found in photoreceptors of the bovine eye, were immunochemically studied in retina of the new Pleurodeles waltl. Polyclonal monospecific antibodies against recoverin were raised and the methods of immunoblotting and indirect immunofluorescence were used. A protein with an apparent molecular mass of 26 kDa was found in the retina extract, which was specifically stained by the antibodies against recoverin. Localization of recoverin was studied on the retina sections: an intense reaction was found in the inner segments and a weak reaction was found in the basal part of the outer segments of photoreceptors and in Landolt's clubs of displaced bipolars. The results we obtained suggest for the first time the presence of recoverin in the retina of a representative of the Urodeles and indicate to interspecific conservativeness of this protein and differences of its localization in the retina photoreceptors in different species. The data obtained open a possibility of using recoverin as a marker protein of photoreceptors and displaced bipolars in studied of retina regeneration in newts.

Animals↗

Cellular pattern formation in the retina: retinal regeneration as a model system.

Like many structures in the central nervous system, the neural retina is highly organized at the cellular level. Examples of this cellular organization include the laminar profile of the vertebrate retina, the hexagonal array of ommatidia in the retinas of insects, and non-random two-dimensional patterns of specific vertebrate retinal neurons. These organized cellular ensembles are taxonomically robust, and their importance in visual processing is, although typically not well understood, virtually axiomatic. The presence of non-random cellular patterns in the retina also begs questions concerning the spatial nature of the patterns, and the underlying mechanisms that coordinate their assembly during retinal development and growth. What are the spatial characteristics of the non-random cellular patterns? What molecular signaling schemes might account for their assembly? What are good model systems for investigating these issues? In this review we attempt to provide some preliminary answers to these questions. We present recent advances in our understanding of cellular patterns in the vertebrate retina and the mechanisms that underlie their assembly, the ability of adult anamniote retinas to regenerate following injury, and how these seemingly disparate topics can be successfully merged into an effort to better understand both processes. We combine insights from retinal assembly mechanisms in Drosophila with empirical, quantitative, and theoretical investigations in vertebrates, to propose an inclusive model for retinal cell patterning.

Animals↗

Retinal pigment epithelial cell distribution in central retina of rhesus monkeys.

PURPOSE: To determine the cell density profile of the retinal pigment epithelium (RPE) in the central retina and relate it to the distribution of photoreceptors. METHODS: Wholemounts of rhesus monkey (Macaca mulatta) retinas with the choroid removed but the RPE attached were stained with the nuclear stain 4',6-diamidine-2-phenylindole dihydrochloride (DAPI) and imaged with a fluorescence microscope. RPE cell nuclei were counted at the foveal center and at 0.4-mm intervals along the vertical meridian. The number of photoreceptors per RPE cell at each location was estimated by using previously published data on the distribution of rhesus photoreceptors. RESULTS: Data were collected from eight retinas. Mean RPE cell density increased from a relatively stable baseline of approximately 4000 RPE cells/mm(2) beyond 2 mm (10 degrees ) eccentricity to more than 7000 cells/mm(2) at the center of the fovea. The number of cones per RPE cell in the rhesus retina was approximately 20:1 in the foveal center (similar to the human retina) and only approximately 1.5:1 in the parafovea. However, when the rods were included, and the total number of photoreceptors per RPE cell were considered, the ratio of photoreceptors to RPE cells was lower in the fovea than in the remainder of the central retina. CONCLUSIONS: In spite of the high cone density, there is a relatively low number of photoreceptors per RPE cell in the fovea. This may limit the metabolic demands on foveal RPE cells and help to preserve their functions in the face of aging and disease.

Animals↗

Retarded outer segment development in TrkB knockout mouse retina organ culture.

PURPOSE: To determine the effects of trkB deficiency in the mouse retina on photoreceptor development and retinal organization, in the absence of confounding systemic effects. METHODS: Newborn mice that carried two null trkB alleles (trkB-/-) and their wild type (WT) littermates were used for retinal organ cultures. On Day 21, rod development was assessed histologically in plastic sections (outer segment length) and retinal organization was analyzed using retinal cell-type specific antibodies. Anatomical data obtained from the organ cultures were compared to previously published histological results from in vivo data. RESULTS: (1) Rod outer segment length was significantly shorter in retinas from trkB-/- mice in the presence of normal numbers of rods. (2) No dopaminergic amacrine cells were observed in the knockout retina. (3) Unlike in the in vivo condition, recoverin-positive OFF-cone bipolar cells were present in trkB-/- retinas grown in culture. CONCLUSIONS: (1) These results demonstrate that rod outer segment development is compromised in the absence of trkB in the retina. (2) This study further supports our previous conclusion that the elimination of trkB expression alters rod development, because the presence of trkB receptors within the retina is essential for normal rod maturation and not because of confounding systemic effects. (3) More generally, this study stresses the importance of investigating complex phenotypes in gene knockout mice under conditions that isolate the organ under investigation from unrelated systemic variations.

Animals↗

[Expression of regulatory homeobox genes during retina regeneration in adult newts].

We studied molecular-genetic mechanisms of retina regeneration in amphibians and, specifically, expression of the homeobox genes Pax6, Prox1, and Six3 in normal development and during retina regeneration in the newt. Based on the structural analysis of genes in closely related amphibian species, primers were constructed that flank certain regions of these genes. PCR fragments of calculated length were obtained. The affiliation of PCR products to the above genes was confirmed by sequencing. A comparative PCR analysis of expression of Pax6, Prox1, and Six3 was carried out in the native and regenerating newt retina, which allowed estimation of the level of expression. cDNA libraries obtained from the native and regenerating retina were used as templates. The libraries were preliminary standardized according to glyceraldehydes-3-phosphate dehydrogenase, an enzyme of general cell metabolism. The genes we studied were expressed in both native and regenerating retina. The level of Pax6 and Prox1 expression increased during regeneration, while that of Six3 decreased. The decrease in the level of Six3 expression could be due to antagonistic interrelations of Prox1 and Six3. The changed level of Prox1 and Six3 expression is a new fact and requires further studies. The interactions between these and other regulatory genes and localization of their expression in the cells of native and regenerating retina will be studied using in situ hybridization and immunohistochemistry.

Age Factors↗

Serotonin N-acetyltransferase (NAT) induction in mammalian retina: role of cyclic AMP and calcium ions.

In retinas and pineal glands of rat, rabbit and hen, activities of the penultimate (and key regulatory) enzyme in melatonin biosynthesis, serotonin N-acetyltransferase (NAT), display distinct diurnal variations, with high and low values during dark and light phase of a 12-h dark: 12-h light illumination cycle. Two-hour incubation (during daytime hours in light) of isolated pineal glands of the studied vertebrates, or the retinas, with 50 microM forskolin (plus 100 microM 3-isobutyl-1-methylxanthine, IBMX-a phosphodiesterase inhibitor), and 1 mM dibutyryl-cAMP, markedly increased the tissue NAT activity. The same procedures significantly enhanced the enzyme activity of rat retina in light, however, only during nighttime hours. The forskolin (+ IBMX)-induced increase of NAT activity in rat retina was significantly lower in a calcium-free medium, and substantially enhanced when calcium concentration was raised from 1.3 mM to 3.9 mM. Treatment of rats with IBMX or aminophylline, and rabbits with aminophylline, increased NAT activity in their pineal glands irrespective of the time of the day, whereas both phosphodiesterase inhibitors significantly increased the enzyme activity of rat retina only when injected during the subjective dark hours. It is concluded that, by analogy to vertebrate pineal gland, in vertebrate retina an increase of NAT activity (and consequently melatonin formation), stimulated both physiologically (i. e. at night), or pharmacologically, involves a cAMP- and calcium dependent process of the enzyme induction.

1-Methyl-3-isobutylxanthine↗

Study on RNA synthesis in the retina and retinal pigment epithelium of mice by light microscopic radioautography.

With the aim of determining the distribution of the incorporation of 3H-uridine in both retina and retinal pigment epithelium (RPE), the mouse eyes at embryonic day 9.5 (E 9.5), E 12.5, E 14.5, E 16.5, E 18.5 of gestational ages, and postnatal day 1 (P 1), P 3, P 7, P 14 were analyzed by light microscopic radioautography. Small pieces of the ocular tissues were labelled with 3H-uridine in vitro and light microscopic radioautographs were prepared. The average grain numbers per cell of the respective regions of tissues were calculated. In the retina, the grain numbers increased gradually from E 9.5 to P 1 and reached the maximal value at P 1, and then decreased until P 14. However, the grain numbers were more in the vitreal portion than those in the scleral portion at E 16.5 and then became more in the scleral portion from E 18.5 to P 14. It is considered that the ganglion and bipolar cells finish the RNA synthesis earlier, while the photoreceptor cells do it later during the fetal and postnatal development. In the RPE, the grain numbers gradually increased from E 12.5 to P 7 and then decreased until P 14. Considering the same ages, the grain numbers increased in the following order, anterior, equatorial and posterior regions during embryonic stages, but decreased in the same order after birth. Therefore, it is suggested that the activity of RNA synthesis in PE cells is higher in the posterior region than in the anterior region during embryonic stages. But the activity ascends generally and becomes relatively higher in the anterior region, after birth. Comparing the retina and RPE, it was noted that the grain numbers in the RPE were more important than in the retina and that the maximal value was at P 1 in the retina, while it was at P 7 in the RPE. From these results, it can be concluded that the RNA synthesis ceases earlier in the retina than in the RPE.

Aging↗

[Effects of supplementation of different kinds of iodine on the antioxidative ability of retina in iodine deficient rats].

OBJECTIVE: To study the effects of iodine supplementation (in different kinds and doses) on the antioxidative ability of retina in iodine deficient rats. METHODS: One hundred and twenty eight iodine deficient Wistar rats were randomly divided into four groups, normal dose of iodate, normal dose of iodide, high dose of iodate and high dose of iodide. Concentration of serum thyroid hormones, including total TT(3) and total TT(4), were estimated by radioimmunoassay. GSH-Px, SOD, TAOC activities and MDA content in the retina were determined using biochemical methods in the 22nd week of iodine supplementation. RESULTS: Statistical analysis showed that a significant difference in TT(3) level of serum was observed between animals treated with different doses. Serum TT(3) level in the groups treated with high doses was significantly higher than those with normal doses. However, no statistical difference could be detected at TT(4) level between animals treated with different doses. Different kind of iodine did not affect the level of thyroid hormones. Statistical analysis showed that a difference in SOD activity of retina was observed between animals treated with different doses. SOD activity in the groups with normal doses was significantly higher than that in groups with larger doses. Retina TAOC activity was significantly higher in groups treated with iodide than that in groups of iodate. Although there was no statistical difference in GSH-Px activity between different groups, it showed the same tendency as the SOD and TAOC activities, i.e. GSH-Px activity in the groups of normal doses was higher than that in the groups of high doses. GSH-Px activity in groups of iodide was higher than that in the groups of iodate. There was no significant difference in MDA content among these four groups. CONCLUSIONS: Different iodine and doses have certain effects on the antioxidative ability of retina in iodine deficient rat. The rats supplemented potassium iodide at normal dose showed higher antioxidative ability of the retina than those of the others.

Animals↗