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Turnover and orientation of the major neural retina cell surface protein protected from tryptic cleavage by calcium.

Calcium protects a limited number of embryonic chick neural retina cell surface proteins from tryptic cleavage. One glycoprotein of Mr approximately 1.3 X 10(5) and pI approximately 4.8 (gp130) is present in intact retinas and, in the presence of Ca2+, is both resistant to tryptic cleavage and poorly iodinated. When cultured in vitro, iodinated retinas release into the medium a number of iodinated polypeptides; one of the major iodinated components is a polypeptide of Mr approximately 9 X 10(4) and pI approximately 4.8 (gp90). This component is also resistant to tryptic cleavage in the presence of Ca2+. Two-dimensional peptide maps of gp130 and gp90 derived respectively from iodinated retinas and their conditioned media are very similar. Maps of samples reiodinated following denaturation show the same similarities as well as additional labeled peptides. Furthermore, the two-dimensional peptide maps of the two molecules prepared from retinas cultured in the presence of [3H]glucosamine are identical. We conclude that gp90 is a turnover fragment of gp130 and comprises that portion of gp130 exposed at the cell surface. The relevance of these polypeptides to Ca2+-dependent retina cell-cell adhesion and their similarity to polypeptides implicated in Ca2+-dependent adhesion of other cell types are discussed.

Animals↗

Identification of oxidatively truncated ethanolamine phospholipids in retina and their generation from polyunsaturated phosphatidylethanolamines.

Oxidized (ox) phospholipids are receiving growing recognition as important messengers in oxidative stress signaling pathways and as endogenous electrophilic toxins that interfere with protein function through covalent modifications. Phosphatidylcholine lipids predominate in low-density lipoproteins (LDL). Our previous studies of oxLDL identified a family of biologically active oxidatively truncated phosphatidylcholines that are also present in atherosclerotic plaques. In contrast, phosphatidylethanolamine (PE) lipids are extraordinarily abundant in retina. Because photoreceptors contain the most highly unsaturated fatty acids found in vertebrate tissues, these membranes are expected to be especially susceptible to oxidative damage. Here, we report that oxidatively truncated ethanolamine phospholipids (oxPEs) are present in retina. As expected, the most abundant oxPEs, succinyl (2.2 +/- 0.8 pmol/retina) and omega-oxobutyryl (1.5 +/- 1.0 pmol/retina) esters of 2-lysophosphatidylethanolamine, are derived from the docosahexaenoyl ester, the most abundant polyunsaturated PE in retina. However, a large amount of the omega-oxononanoyl ester (1.3 +/- 0.6 pmol/retina), derived from linoleyl-PE, is also present even though linoleate is an order of magnitude less abundant than docosahexenoate in retina. There is a notable trend for the presence in retina of greater amounts, relative to the levels of their precursors, of longer chain homologous aldehydes and alkanedioate monoesters. We considered the possibility that this trend results from differences in the proclivities of various polyunsaturated fatty acyl (PUFA)-PEs to generate these homologous products. Therefore, we examined oxidative cleavage of various PUFA-PEs in small unilamellar vesicles. Alkanedioate monoesters are the major stable end products. Particularly notable is the fact that omega-oxononanoyl-PE levels either do not decline or decline less than those of the analogous aldehydes omega-oxobutyryl-PE or omega-oxovaleryl-PE during autoxidation for 33 h. The resistance of omega-oxononanoyl-PE, as compared with omega-oxobutyryl-PE and omega-oxovaleryl-PE, to further oxidation may contribute to the greater amount of this oxPE relative to its precursor, linoleyl-PE, in retina.

Animals↗

Non-neuronal cells involved in the degeneration and regeneration of the fish retina.

In the present work we show that during the degenerative process occurring after the cryo-elimination of the tench peripheral growing zone many non-neuronal cell types in addition to the resident microglial cells, appear within the affected areas. Some of them are normally found in the retina, such as the retinal pigmented epithelium cells and others originate from extra-retinal tissues. We identified these as granular leukocytes and macrophages. The microglial cells and macrophages, those resident in the sub-retinal space, and the invasive ones, act as phagocytes. The analysis of the injured retina following lesion shows that the invasive macrophages, arising from the scleral extra-retinal tissues, penetrate the neural retina, and migrate from the scleral to the vitreal portion. In contrast those coming from the vitreal extra-retinal tissues migrate in the opposite direction. Moreover, the retinal pigmented epithelium cells present remarkable modifications in their morphology and distribution and enter the neural retina, where they disrupt the surrounding tissue. We have also observed that this cryo-lesion causes an inflammation mediated by a type of granular leukocyte, denominated heterophils which penetrate the neural retina and probably come from the blood supply. Our results suggest that, during the first days after the lesion, the participation of diverse non-neuronal cells removing cell debris from the damaged zone should create a favourable environment allowing the regeneration of the neural retina.

Animals↗

The neurons of the ground squirrel retina as revealed by immunostains for calcium binding proteins and neurotransmitters.

Ground squirrel retinas were immunostained with antibodies against calcium binding proteins (CBPs) and classical neurotransmitters in order to describe neuronal phenotypes in a diurnal mammalian retina and to then compare these neurons with those of more commonly studied nocturnal retinas like cats' and rabbits'. Double immunostained tissue was examined by confocal microscopy using antibodies against the following: rhodopsin and the CBPs, calbindin, calretinin, parvalbumin, calmodulin and recoverin (CB, CR, PV, CM, RV), glycine, GABA, choline acetyltransferase (CHAT) and tyrosine hydroxylase (TOH). In ground squirrel retina, the traditional cholinergic mirror symmetric amacrine cells colocalize CHAT with PV and GABA and faintly with glycine. A second cholinergic amacrine cell type colocalizes glycine alone. CR is found in at least 3 different amacrine cell types. The CR-immunoreactive (IR) cell population is a mixture of glycinergic and GABAergic types. The dopamine cell type IR to tyrosine hydroxylase has the typical morphology of a wide field cell with dendrites in S1 but the "rings" seen in cat or rabbit retina are not as numerous. TOH-IR amacrine cells send large club-shaped processes to the outer plexiform layer. CB and CR are in bipolar cells, A- and B-type horizontal cells and several amacrine cell types. Anti-rhodopsin labels the low density rod photoreceptor population in this species. Anti-recoverin labels cones and some bipolar cells while PKC is found in several different bipolar cell types. One ganglion cell with dendritic branching in S3 is strongly CR-IR. We find no evidence for an AII amacrine cell in the ground squirrel, with either anti-CR or anti-PV. An amacrine cell with similarity to the DAP1-3 cell of rabbit is CR-IR and glycine-IR. We discuss this labeling pattern in relationship to other mammalian species. The differences in staining patterns and phenotypes revealed suggest a functional diversity in the populations of amacrine cells according to whether the retinas are rod or cone dominated.

Amacrine Cells↗

Cellular and subcellular localization of heme oxygenase-2 in monkey retina.

Carbon monoxide (CO), an activator of soluble guanylate cyclase (SGC) and generated enzymatically by heme oxygenases (HO), is considered to function as an intra- and intercellular neuromodulator or neurotransmitter in the central and peripheral nervous systems. HO-2 is the constitutive isoform of HO and is more prevalent in nervous tissues than in the other peripheral tissues. Because previous studies have demonstrated different distributions of HO-2 in the retina depending on the species of animals, the aim of this study was to identify which cell types of the monkey retina express HO-2. The expression of HO-2 protein was examined in monkey retina by Western blot analysis. Immunoblottings from monkey homogenates revealed a single clear protein band with a molecular mass of 36 kDa that is corresponding to rat HO-2. Immunoreactivity of HO-2 was found in the perikarya of ganglion cells. Density of immunoreactive ganglion cells was higher in the central area of retina than in the peripheral retina, and somata of larger ganglion cells were stained more densely than smaller ones. In electron microscopy, immunoreactivity of HO-2 was localized on the membrane of the endoplasmic reticulum and the nuclear outer membrane of the ganglion cells. By contrast, inner plexiform layer, inner nuclear layer and outer nuclear layer were devoid of HO-2 immunoreactivity. cGMP were strongly localized in all of ganglion cells. Some cells contributed to the relatively faint cGMP staining were seen in the inner nuclear layer. In combination of HO-2 and cGMP immunocytochemistry, the overlap of co-localization of HO-2 and cGMP would suggest that HO-2 in the ganglion cells would serve as a source for CO generation and CO could serve as a gaseous signaling molecule modulator of neural activity in the retina of monkey.

Animals↗

Pineal control of the dopamine D2-receptor gene and dopamine release in the retina of the chicken and their possible relation to growth rhythms of the eye.

Retinal dopamine (DA) and the DA D2-receptor have been implicated in the development of "deprivation myopia", induced by frosted eye occluders. We have studied the changes in D2-mediated dopaminergic transmission in the retina, their possible relations to eye growth rhythms and myopia, and their control by the pineal gland. (1) We found that the sensitivity of eye growth to retinal image degradation varied over the day. Intermittent periods of normal vision inhibited deprivation myopia more if they occurred in the evening than in the morning. (2) Diurnal growth rhythms in both eyes interacted even though it was previously shown that both deprivation myopia and the accompanying changes in retinal DA release can be monocularly induced. (3) The D2-receptor mRNA concentration in the retina showed no systemic diurnal changes and was not affected by deprivation myopia, but was increased after 2 days in darkness. Since DA release varies over the day, the gain of dopaminergic transmission may also vary, which could explain the observation described in (1) above. (4) Depletion of retinal DA by intravitreal application of reserpine, which lowers DA content severely, had little effect on D2-receptor mRNA concentration. (5) Selective illumination of the pineal gland reduced the D2-receptor mRNA content in the retina to a similar level to full illumination, indicating that the pineal gland controls the D2-receptor mRNA content in the retina. The pineal also controlled DA release in the retina. These results show that the pineal has a surprisingly large influence on both the retinal DA receptor gene transcription and DA release. It can probably control the gain of dopaminergic transmission in the retina and deprivation myopia and mediate the interactions of the growth rhythms in both eyes.

Animals↗

Expression of antiapoptotic and proapoptotic molecules in diabetic retinas.

PURPOSE: To investigate the expression of the antiapoptotic and proapoptotic markers in diabetic retinas. METHODS: In total, 12 donor eyes from six subjects with diabetes mellitus, and 10 eyes from five nondiabetic subjects without known ocular disease serving as control subjects were examined. Immunohistochemical techniques were used with antibodies directed against cyclooxygenase-2 (Cox-2), Akt (protein kinase B), Mcl-1, Bad, cytochrome c, apoptosis-inducing factor (AIF), tumour necrosis factor receptor-1-associated death domain protein (TRADD), and Fas-associated death domain protein (FADD). RESULTS: In retinas from all subjects without diabetes, cytoplasmic immunoreactivity for the antiapoptotic molecules Cox-2, Akt, and Mcl-1 was noted in ganglion cells. Cytoplasmic immunostaining for Cox-2 was also noted in the retinal pigment epithelial cells. Weak immunoreactivity for the mitochondrial apoptogenic proteins cytochrome c, and AIF was noted in the inner segments of photoreceptors, in the inner one-third of the outer plexiform layer, in cells in the inner nuclear layer, in the inner plexiform layer, and in ganglion cells. There was no immunoreactivity for the other antibodies tested. All diabetic retinas showed de novocytoplasmic immunoreactivity for Bad in ganglion cells, and in occasional cells in the inner nuclear layer. Upregulation of cytochrome cand AIF immunoreactivity was noted. Cox-2, Akt, and Mcl-1 immunoreactivity was not altered in the diabetic retinas. There was no immunoreactivity for TRADD, and FADD. CONCLUSIONS: Ganglion cells in diabetic and nondiabetic retinas express the antiapoptotic molecules Cox-2, Akt, and Mcl-1. Retinal ganglion cells express the proapoptotic molecule Bad in response to diabetes-induced neuronal injury. Diabetic retinas show upregulation of the mitochondrial proteins cytochrome c, and AIF.

Aged↗

Active synthesis of C24:5, n-3 fatty acid in retina.

The formation of 14C-labelled long-chain and very-long-chain (n-3) pentaenoic and hexaenoic fatty acids was studied in bovine retina by following the metabolism of. [14C]-docosapentaenoate [C22:5, n-3 fatty acid (22:5 n-3)], [14C]-docosahexaenoate (22:6 n-3), and [14C]acetate. With similar amounts of 22:5 n-3 and 22:6 n-3 as substrates, the former was actively transformed into 24:5 n-3, whereas the latter was virtually unmodified. Labelled 24:5, 26:5, 24:6 and 22:6 were formed from [1-14C]22:5 n-3, showing that pentaenoic fatty acids including 24:5 n-3 can be elongated and desaturated within the retina. When retinal microsomes were incubated with [1-14C]22:5 n-3, 24:5 n-3 was the only fatty acid formed. In retinas incubated with [14C]acetate, 24:5 n-3 was the most highly labelled fatty acid among the polyenes synthesized, 24:6 n-3 being a minor product. Such selectivity in the elongation of two fatty acids identical in length, 22:5 n-3 and 22:6 n-3, despite the fact that 22:5 is a minor and 22:6 a major fatty acid constituent of retina, suggests that the active formation of 24:5 n-3 plays a key role in n-3 polyunsaturated fatty acid (PUFA) metabolism. This compound might give rise to even longer pentaenes via elongation, and to the major PUFAs of retina, 22:6 n-3, by 6-desaturation and chain shortening. Of all retinal lipids, a minor component, triacylglycerol (TG), incorporated the largest amounts of [14C]22:5 and 22:6. TG also concentrated most of the [14C]24:5 formed in retina, whether from [14C]22:5 n-3 or from [14C]acetate, suggesting an important role for this lipid in supporting PUFA metabolism and the synthesis of 22:6 n-3.

Acetates↗

Blood supply to the retina and the lens in the gerbil (Meriones unguiculatus).

The blood supply to the retina and the lens in 32 gerbils (Meriones unguiculatus) of both sexes from infancy to maturity was studied under light and stereoscopic microscopes, and a scanning electron microscope. Mercox (CL-2R; Dai Nippon Ink, Tokyo, Japan) was injected into the left ventricle of 30 animals in order to visualize the blood supply to the retina and the lens from the ophthalmic artery. The central retinal artery arises from the ophthalmic artery, passes through the papilla of the optic nerve together with the central retinal vein and penetrates the vitreous space (cavity of the eye) between the lens and the internal limiting membrane of the retina, where it divides into the central branches covering the lens and the parietal branches to supply the retina. The former passes through the hyaloid space after branching several arterioles and then covers the lens like a network from its medial and marginal sides. Different from small experimental animals, the parietal branches, just after separating from the central one, divides into the nasal, dorsal and temporal branches in the vitreous space, each of which then subdivides to distribute across the retina on the inner limiting membrane, then delineates the membrana vasculosa retinae. This basal pattern of vasculization 1 day after birth continues to death. Both the central and parietal branches of the central retinal artery correspond to the branches of the hyaloid artery in embryo and the latter is preserved in adult gerbils.

Animals↗

Melatonin effect on [3H] glutamate uptake and release in the golden hamster retina.

The effect of melatonin on [3H]glutamate uptake and release in the golden hamster retina was studied. In retinas excised in the middle of the dark phase, i.e., at 2400 h, melatonin (0.1 and 10 nM) significantly increased [3H]glutamate uptake, and this effect persisted in a Ca2+-free medium. On the other hand, melatonin significantly increased [3H]glutamate release in retinas excised at 2400 h, but this effect was Ca2+ sensitive. Melatonin significantly increased 45Ca2+ uptake by a crude synaptosomal fraction from retinas of hamsters killed at 2400 h. In retinas excised at 1200 h, melatonin had no effect on [3H]glutamate uptake, [3H]glutamate release, or 45Ca2+ uptake at any concentration tested. Cyclic GMP analogues, i.e., 8-bromoguanosine 3',5'-cyclic monophosphate and 2'-O-dibutyrylguanosine 3',5'-cyclic monophosphate, significantly increased [3H]-glutamate uptake, [3H]glutamate release, and 45Ca2+ uptake by tissue removed at 1200 and 2400 h, suggesting that the effects of melatonin could correlate with a previously described effect of melatonin on cyclic GMP levels in the golden hamster retina. Taking into account the key role of glutamate in visual mechanisms, the results suggest the participation of melatonin in retinal physiology.

Animals↗

Developmental alteration of the expression and kinase activity of cyclin-dependent kinase 5 (Cdk5)/p35nck5a in the rat retina.

Neuronal Cdc2-like kinase has been purified from the bovine brain as a proline-directed serine/threonine kinase. This kinase is a heterodimer of Cdk5 and p35nck5a and influences neuronal maturation or sprouting in the normal brain. In this study, we showed the expression of Cdk5/p35nck5a kinase in the developing rat retina. The expression of Cdk5 and p35nck5a increased between 1 week and 3 weeks after birth. These expression levels were most prominent from 2 weeks to 3 weeks after birth and decreased after 4 weeks. The developmental change of Cdk5/p35nck5a kinase activity coincided with those of the expression of p35nck5a and Cdk5. An immunohistochemical study showed that Cdk5 was expressed in the ganglion cells and in some cells in the inner nuclear layer at an early stage. With retinal development, Cdk5 was expressed in the inner plexiform layer also. In the adult, the expression of Cdk5 was restricted to the inner plexiform layer and to some cells in the inner nuclear layer. These changes of localization in the developing retina were very close to those of B-50/GAP-43. On the other hand, the expression of p35nck5a was restricted to the soma of the neuron in the developing retina. This subcellular localization in the developing retina agreed with that in the developing rat brain. The expression levels of Cdk5 and p35nck5a in retina of rats raised from fetus to 3 weeks after birth in darkness were 36% and 40% respectively, of the baseline for control rats. Moreover, the kinase activity in rats raised in darkness was lower than that in control rats. These data suggest that Cdk5/p35nck5a may play a role in neuronal plasticity in the developing rat retina.

Animals↗

Rhythmic changes in metabolism of dopamine in the chick retina: the importance of light versus biological clock.

Rhythmic changes in dopamine (DA) content and metabolism were studied in retinas of chicks that were adapted to three different lighting conditions: 12-h light : 12-h dark (LD), constant darkness (DD) and continuous light (LL). Retinas of chicks kept under LD conditions exhibited light-dark-dependent variations in the steady-state level of DA and the two metabolites of DA, i.e. 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanilic acid (HVA). Concentrations of DA, DOPAC and HVA were high in light hours and low in dark hours of the LD illumination cycle. In retinas of chicks kept under DD, the content of DA, DOPAC and HVA oscillated in a rhythmic manner for 2 days, with higher values during the subjective light phase than during the subjective dark phase. The amplitudes of the observed oscillations markedly and progressively declined compared with the amplitudes recorded under the LD cycle. In retinas of chicks kept under LL conditions, levels of DA, DOPAC and HVA were similar to those found during the light phase of the LD cycle. Changes in the retinal contents of DA and HVA did not exhibit pronounced daily oscillations, while on the first day of LL the retinal concentrations of DOPAC were significantly higher during the subjective light phase than during the subjective dark phase. Acute exposure of chicks to light during the dark phase of the LD cycle markedly increased DA and DOPAC content in the retina. In contrast, light deprivation during the day decreased the retinal concentrations of DA and DOPAC. It is suggested that of the two regulatory factors controlling the level and metabolism of DA in the retina of chick, i.e. light and biological clock, environmental lighting conditions seem to be of major importance, with light conveying a stimulatory signal for the retinal dopaminergic cells.

3,4-Dihydroxyphenylacetic Acid↗

Effect of gestational nicotine treatment on newborn rat retina: a histopathological and morphometric analysis.

BACKGROUND: Smoking is a significant risk factor in several debilitating and fatal diseases. It has been implicated in bilateral tobacco-toxic and Leber's hereditary optic neuropathies. Although it has been demonstrated that smoking has a cumulative effect on retinal and optic nerve functions and causes diffuse and localised retinal sensitivity decrease in healthy chronic heavy smokers, the affected retinal layer has not been identified and there is no experimental study investigating the effect of nicotine exposure during gestation on the newborn rat retina. PURPOSE: This experimental investigation evaluated histologically the influence in vivo of maternal nicotine treatment during pregnancy on the newborn rat retina. Different dosages of the test compound simulated the range of low, moderate, and heavy smokers in humans. METHODS: Experimentally naive, adult female Wistar-albino rats weighing 200-250 g were mated with adult male rats over 2 days for copulation in the proportion of two females for every male animal. After confirming pregnancy with vaginal smear method, 40 gravid rats (dams) were then randomly assigned into four equal groups (three experimental and one control; n = 10 in each). On day 9 of gestation, groups 1, 2, and 3 experimental dams were treated with intraperitoneal (i.p.) (-)-nicotine tartrate at doses of 0.5, 1, and 2 mg kg body weight-1 day-1, respectively during pregnancy from gestational day 9-21. Group 4 control dams were given i.p. saline solution daily for the same period. After normal delivery, the newborn litters were sacrificed at postnatal day 1 or day 30. The eyes were enucleated for histopathologic and morphometric analysis of the retinas. Nicotine-induced neuronal changes were measured by morphometric analyses on cell counts of ganglion cell layer (linear cell density in number per unit length of retina) and thickness of the various retinal layers. RESULTS: The litters in control group 4, and experimental groups 1 and 2 had normal retinal findings. On the other hand, morphometric analysis of retinal sections in experimental group 3 eyes demonstrated a 20.7% decrease in the number of surviving ganglion cells (40.7 +/- 2.0) compared with controls (51.3 +/- 1.1; p < 0.001). The thickness of whole retina (126.6 +/- 5.4 microm) was also reduced by 13.5% compared with controls (146.3 +/- 4.5 microm; p = 0.007). The main site of retinal atrophy was the inner plexiform layer (30.1 +/- 1.6 microm vs 43.5 +/- 1.3 microm; p < 0.001) with almost no change in the other retinal layers. CONCLUSIONS: Gestational nicotine treatment induces marked changes in the organisation of the developing retina in newborn rats histopathologically. Quantitative morphometric analysis clearly demonstrated that the two most affected structures were the retinal ganglion cells and the inner plexiform layer, both of which are supplied by central retinal artery.

Animals↗

Application of quantitative competitive polymerase chain reaction for measurements of mRNA from antioxidative enzymes in the diabetic rat retina and kidney.

This present study applied quantitative competitive polymerase chain reaction (QC-PCR) in the analyses of mRNA expression of the endogenous antioxidative enzymes CuZn superoxide dismutase (SOD), MnSOD, catalase, and glutathione peroxidase in tissue samples from the retina and kidney cortex of diabetic rats. RNA was extracted from snap-frozen retinas and pieces of the kidney cortex of male Wistar rats with streptozotocin (STZ)-induced diabetes and control rats. The mRNA levels were analyzed using QC-PCR. The animals were kept in the laboratory for 1 and 6 months, respectively, and fed a normal or probucol- (1% wt/wt) enriched diet. By using QC-PCR, relative mRNA levels of all antioxidative enzymes could be estimated in the retina and kidney cortex. In the retina, the relative catalase mRNA concentration was about 1/10 that of the other enzymes. After 6 months of diabetes, there was a 100% increase of the catalase (median, 0.012 [range, 0.008 to 0.017] v 0.006 [0.005 to 0.010]; P =.011) and a 50% increase of the glutathione peroxidase mRNA levels (0.88 [0.44 to 1.12] v 0.52 [0.31 to 0.79]; P =.044). In the kidney cortex, the relative glutathione peroxidase mRNA level was 10 to 15 times higher, and catalase mRNA level about half of those of CuZnSOD and MnSOD. After 1 month of diabetes, there was an increase only of the glutathione peroxidase mRNA levels, by 170% (17.59 [6.19 to 29.49] v 6.96 [2.34 to 9.04]; P =.047). We conclude that quantification of mRNA can provide difficulties when the amount of sample RNA is limited and/or the gene expression is low. The present study shows QC-PCR to be useful as a tool for measuring expression of mRNA not only in the kidney cortex but also in small tissue samples like the retina. Our results indicate modestly increased mRNA expression of catalase and glutathione peroxidase in the retina and likewise modestly increased mRNA expression of glutathione peroxidase in the kidney cortex of rats with STZ-induced diabetes. Extended studies, also including enzyme activities, are needed before any effect of hyperglycemia on the overall enzyme activity can be established.

Animals↗

Thyrotropin-releasing hormone and somatostatin inhibit each others release in vitro in the rat retina.

The effect of thyrotropin-releasing hormone (TRH), somatostatin (SS) or octreotide, an analogue of SS, on release of TRH or SS from the rat retina was studied in vitro. The retina was incubated in medium 199 (pH 7.4) with 1.0 mg/ml of bacitracin (medium) for 20 min. The amount of TRH or SS release into the medium was measured by individual radioimmunoassays. The TRH release from the rat retina was inhibited significantly in a dose-related manner by the addition of SS or octreotide. The SS release from the retina was inhibited by TRH, and the inhibitory effect of TRH on SS release from the rat retina was blocked by the addition of anti-TRH receptor antiserum immunoglobulin fraction. The findings suggest an interaction between TRH and SS in the rat retina by which the addition of one inhibits the release of the other.

Animals↗

Effects of dopamine on the release of thyrotropin-releasing hormone from the rat retina in vitro.

The effects of dopamine on the release of thyrotropin-releasing hormone (TRH) from the rat retina in vitro were studied. The rat retina was incubated in the medium 199 (pH 7.4) with 1.0 mg/ml of bacitracin and 100 micrograms/ml of ascorbic acid. The amount of TRH release into the medium was measured by radioimmunoassay. The TRH release from the rat retina was inhibited significantly in a dose-related manner with the addition of dopamine, but not with pimozide. The inhibitory effects of dopamine on TRH release from the rat retina were blocked with an addition of pimozide to the medium. The elution profile of methanol-extracted rat retina on sephadex G-10 was identical to that of synthetic TRH. From these findings it is concluded that the dopaminergic system inhibits TRH release from the rat retina in vitro.

Analysis of Variance↗

Effects of serotonin on the release of thyrotropin-releasing hormone from the rat retina in vitro.

Effects of serotonin on the release of thyrotropin-releasing hormone (TRH) from the rat retina were studied in vitro. The retina was incubated in medium 199 (pH 7.4) with 1.0 mg/ml of bacitracin and 100 micrograms/ml of ascorbic acid (medium) for 20 min. The amount of TRH release into the medium was measured by radioimmunoassay. The TRH release from the rat retina was inhibited significantly in a dose-related manner with the addition of serotonin and enhanced with cyproheptadine. The inhibitory effect of serotonin on TRH release from the retina was blocked with the addition of cyproheptadine. The elution profile of methanol extract of the rat retina was identical to that of synthetic TRH. The findings suggest that the serotonergic system inhibits TRH release from the rat retina in vitro.

Animals↗

Profile of the genes expressed in the human peripheral retina, macula, and retinal pigment epithelium determined through serial analysis of gene expression (SAGE).

We used the serial analysis of gene expression (SAGE) technique to catalogue and measure the relative levels of expression of the genes expressed in the human peripheral retina, macula, and retinal pigment epithelium (RPE) from one or both of two humans, aged 88 and 44 years. The cone photoreceptor contribution to all transcription in the retina was found to be similar in the macula versus the retinal periphery, whereas the rod contribution was greater in the periphery versus the macula. Genes encoding structural proteins for axons were found to be expressed at higher levels in the macula versus the retinal periphery, probably reflecting the large proportion of ganglion cells in the central retina. In comparison with the younger eye, the peripheral retina of the older eye had a substantially higher proportion of mRNAs from genes encoding proteins involved in iron metabolism or protection against oxidative damage and a substantially lower proportion of mRNAs from genes encoding proteins involved in rod phototransduction. These differences may reflect the difference in age between the two donors or merely interindividual variation. The RPE library had numerous previously unencountered tags, suggesting that this cell type has a large, idiosyncratic repertoire of expressed genes. Comparison of these libraries with 100 reported nonocular SAGE libraries revealed 89 retina-specific or enriched genes expressed at substantial levels, of which 14 are known to cause a retinal disease and 53 are RPE-specific genes. We expect that these libraries will serve as a resource for understanding the relative expression levels of genes in the retina and the RPE and for identifying additional disease genes.

Adult↗