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Independent guidance of retinal axons in the developing visual system of Drosophila.

The development of the adult visual system of Drosophila requires the establishment of precise retinotopic connections between retinal photoreceptor cell axons and their synaptic partners in the optic lobe of the brain. To assess the role of axon-axon interactions in retinal axon guidance, we used genetic methods to disrupt the normal spatiotemporal order of retinal axon ingrowth. We examined retinal axon projections to the developing first optic ganglion, the lamina, in two mutants in which reduced numbers of ommatidia develop in the eye imaginal disk. We find that in the developing lamina of these mutants, sine oculis and Ellipse, retinal axons project to proper dorsoventral positions despite the absence of the usual array of neighboring retinal axons. In a second approach, we examined animals that were somatic mosaics for the mutation, glass. In glass- animals, retinal axons project aberrantly and the larval optic nerve is absent. We find that in the developing lamina of glass mosaic animals, wild-type retinal axons project to proper dorsoventral positions despite the misrouted projections of neighboring glass- retinal axons. In addition, wild-type retinal axons project normally in the absence of the larval optic nerve, indicating that the latter is not an essential pioneer for retinal axon navigation. Our observations support the proposal that axon fascicles can make at least some pathfinding decisions independently of other retinal axon fascicles. We suggest that positional guidance cues that might label axon pathways and target destinations contribute to retinotopic pattern formation in the Drosophila visual system.

Animals↗

Treatment of symptomatic retinal breaks.

Forty-three patients with symptomatic retinal breaks were treated with photocoagulation and a comparable group of 43 patients were treated with cryopexy. All patients had either a vitreous hemorrhage, or photopsia and floaters. Most of the breaks were of the horseshoe type. Of the patient eyes, 14% (six patients) in both groups developed new retinal tears without detachment during the long term postoperative observation. The new breaks were treated with the method used for the original breaks, and no scleral surgery was necessary. The incidence of retinal detachment was 7% in patients treated with photocoagulation and 11.6% in patients with cryocoagulation, but two cases that developed retinal detachment arising from new tears years after the initial procedure were included.

Cryosurgery↗

Retinal detachment after posterior segment intraocular foreign body injuries.

PURPOSE: To identify the risk factors for retinal detachment after posterior segment intraocular foreign body (IOFB) injuries and to study the association between the development of retinal detachment and visual outcome. METHODS: Ninety-six consecutive patients with posterior segment IOFB injuries were retrospectively reviewed. Vitrectomy techniques were used in primary and secondary treatment. Two eyes were eviscerated after primary repair because of Clostridium perfringens endophthalmitis. Factors analyzed included (1) entrance wound location, (2) presence of uveal prolapse, (3) presence of vitreous prolapse, (4) presence of traumatized iris, (5) presence of endophthalmitis, (6) location of IOFB, (7) size of IOFB, (8) use of scleral buckling and/or an encircling band, (9) use of gas tamponade, (10) use of lensectomy. Data were analyzed using univariate and multivariate logistic regression analysis. RESULTS: Retinal detachment was present in 6 eyes at presentation and occurred in another 19 eyes after vitrectomy. After a mean follow-up of 8.6 months, 63 (65.6%) eyes achieved visual acuities of 20/200 or better, and total retinal detachment complicated by inoperable proliferative vitreoretinopathy was present in 9 (9.4%) eyes. Multivariate analysis identified retinal detachment as a factor significantly associated with a poor visual outcome (odds ratio = 4.54, 95% confidence interval [CI] = 1.05-19.6). Foreign body size of more than 4 mm (odds ratio = 5.8, 95% CI = 1.66-2.03) and presence of endophthalmitis (odds ratio = 11.7, 95% CI = 2.57-52.9) were identified as the only predictive factors for the development of retinal detachment after vitrectomy. Use of prophylactic scleral buckling and/or an encircling band reduced the risk of developing postoperative retinal detachment. CONCLUSIONS: Retinal detachment after posterior segment IOFB injuries is associated with a poor visual outcome. Large IOFB and presence of endophthalmitis are the strongest predictive factors for the development of retinal detachment.

Adolescent↗

Stable properties of spontaneous EPSCs and miniature retinal EPSCs during the development of ON/OFF sublamination in the ferret lateral geniculate nucleus.

Retinal projections to the lateral geniculate nucleus (LGN) in ferrets progressively segregate into eye-specific laminae and subsequently into sublaminae that receive inputs from either ON-center or OFF-center afferents. To study the development of synaptic efficacy during a period of activity-dependent growth and reorganization in the CNS, we recorded spontaneous EPSCs (sEPSCs) from cells of the LGN during ON/OFF sublamination. We also examined retinal inputs specifically by stimulating the optic tract in the presence of strontium and recording evoked miniature EPSCs (emEPSCs). The rise times, areas, half-widths, and decay times of sEPSCs and emEPSCs and interevent intervals of sEPSCs recorded at the beginning of ON/OFF sublamination were not different from those recorded after its completion. Typically EPSC areas were small (10-20 fC) but varied greatly both within and between neurons. The frequency of sEPSCs was also quite variable, ranging from 0.2 to 5 Hz. sEPSCs were equivalent to miniature EPSCs recorded in the presence of tetrodotoxin, and both sEPSCs and emEPSCs were CNQX-sensitive. No difference was observed between sEPSCs recorded at room temperature and those recorded at 34 degreesC, and strontium could be substituted for calcium with no effect on sEPSC shape. These data argue for a remarkable stability in the components of at least AMPA-mediated synaptic transmission during a period of major synaptic rearrangement in the LGN.

6-Cyano-7-nitroquinoxaline-2,3-dione↗

Structural changes in retinal pigmented epithelium of Rivulus marmoratus Poey embryos during development.

In Rivulus marmoratus development of retinal pigmented epithelium (RPE) parallels that of retinal photoreceptors. Although structurally functional by mid-incubation the full complex structure is not achieved even when the yolk-sac is absorbed (3-days post-hatched). Melanogenesis is evident at 0.2 incubation with premelanosomes present up to three days after hatching. The distribution of junctional complexes, basal membrane foldings and coated-pits throughout development is noted. Myeloid bodies, already present at mid-incubation, appear initially as single lenticular-shaped structures which later may form whorls, or coalesce around oil droplets, glycogen clusters or phagosomes thereby giving rise to myeloid patterns characteristic of a mature RPE. The functional significance of these changes is discussed.

Animals↗

N,N-dimethyladriamycin for treatment of experimental proliferative vitreoretinopathy: efficacy and toxicity on the rabbit retina.

N,N-Dimethyladriamycin (Me-2-ADM) derives from adriamycin (doxorubicin) by N-methylation of its aminosugar moiety. In contrast to the parent agent, Me-2-ADM lacks mutagenicity and carcinogenicity. To evaluate its suitability for treatment of proliferative vitreoretinopathy (PVR), we studied the ability of Me-2-ADM to prevent traction retinal detachment in a model of PVR in rabbits. The model was created by intravitreal injection of 25,000 homologous dermal fibroblasts after vitreous gas compression. Two days after fibroblast injection, 5, 10 or 30 nmol Me-2-ADM was administered into the vitreous cavity. The control group received sham injections. All control eyes developed traction retinal detachments. Administration of 5 nmol Me-2-ADM slightly reduced the rate of retinal detachments to 90%. No retinal detachments occurred in the group treated with 10 nmol Me-2-ADM while 11% of eyes treated with 30 nmol Me-2-ADM developed retinal detachments. At day 28 of the study, light microscopic examination of retinas treated with 30 nmol Me-2-ADM revealed severe retinal damage while no such damage was present in eyes treated with 10 nmol. To exclude early retinal damage, 10 nmol Me-2-ADM were injected into eyes that had undergone vitreous gas compression but no fibroblast injection. On day 3, 7, and 14 after drug administration, ERGs were recorded simultaneously from drug treated and sham treated (contralateral) eyes. Light microscopy and transmission electron microscopy (TEM) were performed. Only transient ERG changes were observed at day 7, which recovered by day 14. All morphological findings in drug exposed eyes were in the range of normal when compared with controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Coats' disease of the retina (unilateral retinal telangiectasis) caused by somatic mutation in the NDP gene: a role for norrin in retinal angiogenesis.

Coats' disease is characterized by abnormal retinal vascular development (so-called 'retinal telangiectasis') which results in massive intraretinal and subretinal lipid accumulation (exudative retinal detachment). The classical form of Coats' disease is almost invariably isolated, unilateral and seen in males. A female with a unilateral variant of Coats' disease gave birth to a son affected by Norrie disease. Both carried a missense mutation within the NDP gene on chromosome Xp11.2. Subsequently analysis of the retinas of nine enucleated eyes from males with Coats' disease demonstrated in one a somatic mutation in the NDP gene which was not present within non-retinal tissue. We suggest that Coats' telangiectasis is secondary to somatic mutation in the NDP gene which results in a deficiency of norrin (the protein product of the NDP gene) within the developing retina. This supports recent observations that the protein is critical for normal retinal vasculogenesis.

Amino Acid Substitution↗

Expression of chondroitin sulfate and keratan sulfate proteoglycans in the path of growing retinal axons in the developing chick.

Previous investigations have identified proteoglycans in the central nervous system during development and have implicated some proteoglycans as axon guidance molecules that act by inhibiting axon extension. The present study investigated the pattern of immunoreactivity for several glycosaminoglycans common to certain proteoglycans relative to growing retinal axons in the developing chick visual system and in retinal explant cultures. Immunostaining for chondroitin-6-sulfate, chondroitin-4-sulfate, and keratan sulfate was observed to colocalize with retinal axons throughout the retinofugal pathway during the entire period of retinal axon growth. The proteoglycan form of collagen IX, however, was only observed in the retina, primarily peripheral to the areas with actively growing axons. The pattern of immunostaining for chondroitin sulfate in tissue sections suggested that the retinal axons might be a source for some of the chondroitin sulfate immunostaining in the developing visual pathway. This was confirmed in that chondroitin sulfate immunostaining was also observed on neurites emanating from cultured retinal explants. These findings indicate that retinal axons grow in the presence of chondroitin sulfate and keratan sulfate proteoglycans and that these proteoglycans in the developing chick visual pathway have functions other than to inhibit axon growth.

Animals↗

Dendritic remodelling of retinal ganglion cells during development of the rat.

Investigation of the morphology of ganglion cells in the cat retina has shown that a remarkable reduction in the number of dendritic spines and branches occurs during development of the alpha and beta cell classes. To learn whether dendritic remodelling represents a generalized mechanism of mammalian retinal ganglion cell development, we have examined the morphology of ganglion cells in the retina of the developing rat. The present study has concentrated on type II cells, which retain a great number of dendritic spines and branches in the adult and comprise a large proportion of the population of rat retinal ganglion cells. To reveal fine dendritic and axonal processes, Lucifer yellow was injected intracellularly in living retinae maintained in vitro. Size and complexity of the dendritic trees were found to increase rapidly during an initial stage of development lasting from late fetal life until approximately postnatal day 12 (P12). Dendrites and axons of immature ganglion cells expressed several transient morphological features comprising an excessive number of dendritic branches and spine-like processes, and short, delicate axonal sidebranches. The following developmental stage was characterized by a remarkable decrease in the morphological complexity of retinal ganglion cells and a slowed growth of their dendritic fields. The number of dendritic branches and spines of types I and II retinal ganglion cells declined after P12 to reach a mature level by the end of the first postnatal month. Thus, even cells that retain a highly complex dendritic tree into the adult state undergo extensive remodelling. These results suggest that regressive modifications at the level of the dendritic field constitute a generalized mechanism of maturation in mammalian retinal ganglion cells.

Animals↗

Retinal pigment epithelial tear following intravitreal pegaptanib sodium.

PURPOSE: To report two cases of a retinal pigment epithelial tear after intravitreal injection of pegaptanib sodium. To our knowledge, this is the first report of this finding after intraocular antivascular endothelial growth factor therapy. DESIGN: Observational case reports. METHODS: Two patients presented with occult choroidal neovascularization and associated serous pigment epithelial detachment that was a result of age-related macular degeneration. Both patients were treated with an intravitreal injection of pegaptanib sodium. RESULTS: One patient developed a retinal pigment epithelium tear one week after the intravitreal injection. The second patient developed a retinal pigment epithelium tear eight weeks after treatment. CONCLUSIONS: Although these cases may represent natural history, there should be a high index of suspicion for retinal pigment epithelium tears in patients who report significant visual deterioration after intravitreal injection of pegaptanib sodium. Further studies are needed to determine whether angiographic subtypes of choroidal neovascular membranes are more susceptible to developing retinal pigment epithelium tears after treatment with antivascular endothelial growth factor agents.

Aged↗

Characteristics of sixty myopic eyes with pre-laser in situ keratomileusis retinal examination and post-laser in situ keratomileusis retinal lesions.

PURPOSE: A survey of eyes with pre-laser in situ keratomileusis (LASIK) retinal examinations and characteristics of post-LASIK retinal breaks and retinal detachments (RDs). METHODS: A survey of worldwide vitreoretinal surgeons (424 physicians). Surveyed information included demographics, best-corrected visual acuity, degree of myopia, pre- and post-LASIK retinal findings, follow-up time, and treatment outcome. RESULTS: Sixty eyes with pre-LASIK retinal examinations developed post-LASIK retinal breaks and RDs. There was an average of 2.3 breaks per eye, yielding a total of 140 breaks in the 60 eyes. Forty eyes also had RDs besides the retinal breaks. Large percentages of eyes had substantial myopia (mean myopia, -9.5 +/- 5.8 diopters [D]) and complex vitreoretinal complications. Forty percent developed vitreoretinal complications within 6 months after LASIK. The 20 eyes that developed more extensive RDs (>3 clock hours) had a significantly higher mean myopia than did the 6 eyes that developed limited RD (< or = 3 clock hours) within 12 months after LASIK (-8.92 +/- 6.82 D versus -3.50 +/- 1.97 D, P = 0.03). There were significant statistical differences in distribution of retinal breaks and tears between the temporal and nasal quadrants (P < 0.001, P < 0.001, respectively, chi2, but not between the superior and inferior quadrants. CONCLUSION: Distributions of retinal breaks in this study were comparable with results found in non-LASIK eyes in young myopes. Treatment for post-LASIK vitreoretinal complications was highly successful. The vulnerability of such highly myopic eyes for vitreoretinal complications warrants their close monitoring.

Adolescent↗

Development of a silicon retinal implant: cortical evoked potentials following focal stimulation of the rabbit retina with light and electricity.

There are currently many experimental efforts to develop a treatment for retinitis pigmentosa (RP) and age-related macular degeneration (AMD). The efforts reported here are part of the Retinal Implant Project, a collaborative effort of the Massachusetts Eye and Ear Infirmary and the Massachusetts Institute of Technology, to develop an independent functioning epiretinal prosthesis to restore vision in patients with disease of the outer retina such as RP and AMD. The epiretinal prosthesis will electrically stimulate the inner retina, bypass the damaged photoreceptors, and hopefully result in meaningful vision. One way to monitor the epiretinal implant is to record focal (stimulation of a small area of retina) evoked cortical potentials (EEP) upon electrical current stimulation. EEP recordings have 3 uses: (1) verify that focal retinal electrical stimulation sends a signal to the cortex that is similar to that elicited by light; (2) develop an animal model to chronically monitor the epiretinal implant; and (3) investigate optimal stimulus parameters. Five dutch pelted rabbits were placed under anesthesia and a 250 microm concentric bipolar stimulating electrode was introduced on the vitreal surface of the retina under visual guidance to record the EEP. Stimuli were charge-balanced pulses and recording electrodes were extradural. Focal VEPs were also recorded and compared to the focal EEP to ascertain cortical origin of the EEP, determine similarities of the EEP to the VEP and determine focality of electrical stimulus. EEP recordings were elicited in 3 out of the 5 rabbits. Current amplitudes which produced detectable responses ranged from 50-1000 microA. In our best series, amplitudes increased by 47-206 microV in increasing current from 50-600 microA. In comparing latencies of the focal EEP and focal VEP, the EEP latencies were 60-70 ms faster, favoring a cortical signal origin. In comparing amplitudes, VEP and EEP responses behaved similarly with changes in stimulus strength and stimulating frequency. In conclusion, EEPs were obtained with parameters of electrode size and current threshold level that would be used by the epiretinal implant, enabling use of the EEP response to monitor the epiretinal implant.

Animals↗

Competitive interactions between retinal ganglion cells during prenatal development.

In the mammalian visual system, retinal ganglion cell axons terminate within the LGN in a series of alternating eye-specific layers. These layers are not present initially during development. In the cat they emerge secondarily following a prenatal period in which originally intermixed inputs from the two eyes gradually segregate from each other to give rise to the characteristic set of layers by birth. Many lines of evidence suggest that activity-dependent competitive interactions between ganglion cell axons from the two eyes for LGN neurons play an important role in the final patterning of retinogeniculate connections. Studies of the branching patterns of individual ganglion cell axons suggest that during the period when inputs from the two eyes are intermixed, axons from one eye send side branches into territory later occupied exclusively by axons from the other eye. Ultrastructural studies indicate that these branches in fact are sites of synaptic contacts, which are later eliminated since the side branches disappear as axons form their mature terminal arbors in appropriate territory. In vitro microelectrode recordings from LGN neurons indicate that they can receive convergent synaptic excitation from electrical stimulation of the optic nerves before but not after the eye-specific layers form, suggesting that at least some of the synaptic contacts seen at the ultrastructural level are functional. Finally, experiments in which tetrodotoxin was infused intracranially during the two week period during which the eye-specific layers normally form demonstrate that it is possible to prevent, or at least delay, the formation of the layers. Accordingly, individual axons fail to develop their restricted terminal arbor branching pattern and instead branch widely throughout the LGN. These results indicate that all of the machinery necessary for synaptic function and competition is present during fetal life. Moreover, it is highly likely that neuronal activity is required for the formation of the eye-specific layers. If so, then activity would have to be present in the form of spontaneously generated action potentials, since vision is not possible at these early ages. Thus, the functioning of the retinogeniculate system many weeks before it is put to the use for which it is ultimately designed may contribute to the final patterning of connections present in the adult.

Animals↗

Effects of antibodies to large retinal ganglion cells on developing retinogeniculate pathways in the cat.

We investigated the effects of polyclonal antibodies, produced against ox large retinal ganglion cells, on the developing retinogeniculate pathways of cats. Four-week-old kittens were given an intraocular injection of either a low or a high concentration of the antibodies and effects were assessed 35-69 weeks later. After a low concentration (110 micrograms/33 microliter volume) injection, the density of retinal alpha-cells (the morphological counterpart of Y-cells) was reduced 44% in area centralis and 37% in peripheral retina. After a high concentration (333 micrograms/33 microliter volume) injection, alpha-cell density was reduced 76% in area centralis and 91% in peripheral retina. The same concentration of antibodies had no consistent effect on the numbers of medium- or small-size retinal ganglion cells. Electrophysiological recordings from single neurons in layers A and A1 of the lateral geniculate nucleus (LGN) revealed a 53% decrease in the percentage of Y-cells after a low-concentration injection and an 82% decrease after a high-concentration injection. There was a concomitant increase in the percentage of LGN cells that were non-responsive to light or that responded too poorly to be classified. No change was observed in the percentages of LGN X-cells or cells with mixed response properties. The reduced encounter rate of LGN Y-cells was not accompanied by significant changes in LGN cell-body size. Together, the results indicate that the immunoablation technique produces a large and apparently selective reduction of the Y-cell retinogeniculate pathway in developing kittens.

Animals↗

Visual outcome and prognostic factors after vitrectomy for posterior segment foreign bodies.

PURPOSE: To identify the prognostic factors that predict final visual outcome in eyes with posterior segment intraocular foreign body (IOFB) injuries managed by primary pars plana vitrectomy. METHODS: Ninety-six consecutive patients with posterior segment IOFB injuries were retrospectively reviewed. Factors analyzed included initial visual acuity (VA), time between injury and presentation, site of entrance wound, uveal prolapse, vitreous prolapse, traumatized iris, endophthalmitis, location and size of IOFB, use of scleral buckling and/or an encircling band, gas tamponade, lensectomy, number of surgical procedures, and development of retinal detachment. Data were analyzed using univariate and multivariate logistic regression analysis. RESULTS: After a mean follow-up of 8.6 months, 63 eyes (65.6%) achieved VA of 20/200 or better, and 9 eyes (9.4%) had total retinal detachment complicated by inoperable proliferative vitreoretinopathy. On univariate analysis, predictors of poor vision (hand movements or less) were poor initial VA, corneoscleral entrance wound, uveal prolapse, vitreous prolapse, traumatized iris, and development of retinal detachment. In contrast, predictors of good visual outcome (20/200 or better) were absence of uveal prolapse, no endophthalmitis, and no retinal detachment. Multivariate analysis identified corneoscleral entrance wound, uveal prolapse, and development of retinal detachment as the only factors significantly associated with poor visual outcome. Absence of uveal prolapse was the only factor significantly associated with good visual outcome. CONCLUSIONS: Final visual outcome is greatly determined by the severity of the primary injury. On multivariate analysis, significant predictive factors of final VA were corneoscleral entrance wound, presence or absence of uveal prolapse, and development of retinal detachment.

Adolescent↗

Development of the mouse retinal vasculature: angiogenesis versus vasculogenesis.

PURPOSE: The inner vasculature of the retina develops as a spreading network, which is preceded by spindle-shaped cells. These cells are alleged to be vascular precursor cells (angioblasts). This study was designed to test whether such angioblasts exist in neonatal mouse retina. METHODS: In situ hybridization and immunohistochemistry on mouse retinal wholemount preparations were used to visualize specific vascular cell types. RESULTS: In situ hybridization with an RNA probe against vascular endothelial growth factor receptor (VEGFR)-2 (a marker for endothelial cells and angioblasts) labeled the vascular network but failed to label the spindle-shaped cells in front of it. A probe against VEGFR1, a marker for endothelial cells only, revealed the same staining pattern. Pericytes, visualized with a probe against platelet-derived growth receptor (PDGFR)-beta, were spread over the entire vessel network, but not beyond it. However, in situ hybridization with a probe against PDGFRalpha (a marker for retinal astrocytes) labeled spindle-shaped cells preceding the vessel network. CONCLUSIONS: These observations imply that in the mouse retina the spindle-shaped cells preceding the forming vasculature are immature retinal astrocytes and not vascular precursor cells and that the primary vascular network in the retina develops by angiogenesis (budding from existing vessels) and not vasculogenesis (assembly of dispersed angioblasts).

Animals↗

Localization and quantitation of blood-retinal barrier breakdown in experimental proliferative vitreoretinopathy.

OBJECTIVE: To determine the contribution of the breakdown of the blood-retinal barrier (BRB) as measured with magnetic resonance imaging in the development of retinal detachment in an experimental model of proliferative vitreoretinopathy. METHODS: Contrast-enhanced magnetic resonance imaging was used to evaluate BRB breakdown in an intravitreal cell-injection model of proliferative vitreoretinopathy. Intravitreal injection of 2.5 x 10(5) homologous dermal fibroblasts produced specific disruption of the inner, or vascular, BRB. RESULTS: Breakdown of the BRB was greatest in the first 3 days after injection, confirming previous work using fluorescein-based methods. Injection of 1 mg of intravitreal triamcinolone acetonide at the time of cell injection significantly reduced both BRB breakdown and the incidence of eventual traction retinal detachment. Eyes that did develop detachment had significantly greater leakage prior to its development than those that did not, regardless of steroid treatment. CONCLUSIONS: Quantitation and definitive localization of BRB leakage with magnetic resonance imaging provides a better understanding of the relationship between BRB compromise and the development of retinal detachment in this frequently used model.

Animals↗