Search

Endostatin, a naturally cleaved fragment of type XVIII collagen with antiangiogenic activity, has been involved in the regulation of neovascularization during diabetic retinopathy. Here, the intracellular distribution of endostatin in healthy mouse and human neuroretinas has been analyzed. In addition, to study the effect of experimental hyperglycemia on retinal endostatin, the db/db mouse model has been used. Endostatin protein expression in mouse and human retinas was studied by immunofluorescence and Western blot, and compared with db/db mice. Eye fundus angiography, histology, and immunofluorescence were used to visualize mouse retinal and intravitreal vessels. For the first time, our results revealed the presence of endostatin in neurons of mouse and human retinas. Endostatin was mainly expressed in bipolar cells and photoreceptors, in contrast to the optic disc, where endostatin expression was undetectable. Diabetic mice showed a reduction of endostatin in their retinas associated with the appearance of intravitreal vessels at the optic disc in 50% of db/db mice. Intravitreal vessels showed GFAP positive neuroglia sheath, basement membrane thickening by collagen IV deposition, and presence of MMP-2 and MMP-9 in the vascular wall. All together, these results point that decreased retinal endostatin during experimental diabetes is associated with optic disc intravitreal vascularization. Based on their phenotype, these intravitreal vessels could be neovessels. However, it cannot be ruled out the possibility that they may also represent persistent hyaloid vessels., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Purpose: To determine the underlying reasons for the non-visualization of polyps on en face optical coherence tomography angiography (OCTA) in patients with polypoidal choroidal vasculopathy (PCV)., Methods: A cross-sectional study of consecutive treatment-naïve 30 eyes with active PCV was included. Results of fundus photography, fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA), spectral domain optical coherence tomography (SD-OCT), and en face OCTA were analyzed., Results: A total of 64 active polyps were found on FFA and ICGA in 30 eyes. On OCTA, 42/64 (65.6%) polyps were visualized, while 22/64 (34.4%) polyps were non-visualized. There were no significant differences in the size (P = 0.723) and filling time of polyps (P = 0.558) between the two groups. However, polypoidal lesions were less common in the non-visualized group (P < 0.001). The height of the polyps on SD-OCT was 243.95 ± 114.24 μm in the non-visualized group, which was higher than those (188.00 ± 87.93 μm) in the visualized group (P = 0.048). Moreover, more pulsatile polyps (72.7%) were found in the non-visualized group than those (2.4%) in the visualized group (P < 0.001). Four of the 22 polyps in the non-visualized group (18.2%) were located under a thick subretinal hemorrhage, and two of 22 invisible polyps (9.6%) located under and parallel to the retinal vessel in the inner layer of retina., Conclusions: Our results revealed that the height of the polyps, and not the size and pulsation of the polyps, correlated with the visualization of the polyps on OCTA. Polyps that were pulsating in early ICGA were difficult to be visualized on OCTA, which is the most possible reason for the non-visualization. Coverage with thick subretinal hemorrhage or retina vessels was another reason for the non-visualization of the polyps in active PCV on OCTA.

The development of the ocular vasculatures is perfectly synchronized to provide the nutritional and oxygen requirements of the forming human eye. The fetal vasculature of vitreous, which includes the hyaloid vasculature, vasa hyaloidea propria, and tunica vasculosa lentis, initially develops around 4-6 weeks gestation (WG) by hemo-vasculogenesis (development of blood and blood vessels from a common progenitor, the hemangioblast). This transient fetal vasculature expands around 12 WG by angiogenesis (budding from primordial vessels) and remains until a retinal vasculature begins to form. The fetal vasculature then regresses by apoptosis with the assistance of macrophages/hyalocytes. The human choroidal vasculature also forms by a similar process and will supply nutrients and oxygen to outer retina. This lobular vasculature develops in a dense collagenous tissue juxtaposed with a cell constitutively producing vascular endothelial growth factor (VEGF), the retinal pigment epithelium. This epithelial/endothelial relationship is critical in maintaining the function of this vasculature throughout life and maintaining it's fenestrated state. The lobular capillary system (choriocapillaris) develops first by hemo-vasculogenesis and then the intermediate choroidal blood vessels form by angiogenesis, budding from the choriocapillaris. The human retinal vasculature is the last to develop. It develops by vasculogenesis, assembly of CXCR4 + /CD39 + angioblasts or vascular progenitors perhaps using Muller cell Notch1 or axonal neuropilinin-1 for guidance of semaphorin 3A-expressing angioblasts. The fovea never develops a retinal vasculature, which is probably due to the foveal avascular zone area of retina expressing high levels of antiangiogenic factors. From these studies, it is apparent that development of the mouse ocular vasculatures is not representative of the development of the human fetal, choroidal and retinal vasculatures., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Purpose: A timely regression of the hyaloid vascular system (HVS) is required for the normal ocular development. Although macrophages have a critical role in this process, the exact mechanism remains undetermined. Periostin is a matricellular protein involved in tissue and vascular remodeling. The purpose of our study was to determine whether periostin is involved in the HVS regression., Methods: We used wild type (WT) and periostin knockout (KO) mice. Indocyanine green angiography and immunohistochemistry with isolectin B4 were used to evaluate the HVS regression. TUNEL-labeling was used to quantify the number of apoptotic hyaloid vascular endothelial cells. F4/80 and Iba-1 staining was performed to determine the number and location of macrophages in the vitreous. The location of periostin also was investigated by immunohistochemistry. To determine the functional role of periostin, the degree of adhesion of human monocytes to fibronectin was measured by an adhesion assay., Results: The HVS regression and peak in the number of TUNEL-positive apoptotic endothelial cells were delayed in periostin KO mice. The number of F4/80 positive cells in the vitreous was higher in periostin KO mice. Only a small number of Iba-1-positive cells near the hyaloid vessels was co-stained with periostin, and peripheral blood monocytes were not stained with periostin. Adhesion assay showed that periostin increased the degree of attachment of monocytes to fibronectin., Conclusions: These results suggest that periostin, which is secreted by the intraocular macrophages, enhances the HVS regression by intensifying the adhesion of macrophages to hyaloid vessels.

VEGF(165) b is an anti-angiogenic form of VEGF(165) produced by alternative splicing. The localization of pro-angiogenic VEGF(165) and anti-angiogenic VEGF(165) b was investigated during development of the vasculatures in fetal human eyes from 7 to 21 weeks gestation (WG). The fetal vasculature of vitreous, which includes tunica vasculosa lentis (TVL), had moderate VEGF(165) immunoreactivity at 7WG and very little VEGF(165) b. Both forms were elevated at 12WG. VEGF(165) then decreased around 17WG when the TVL regresses but VEGF(165) b remained elevated. In choroid, VEGF(165) was present in forming choriocapillaris (CC) and retinal pigment epithelium (RPE) at 7WG while VEGF165b was present in CC and mesenchymal precursors within the choroidal stroma. By 21WG, both forms were elevated in RPE and choroidal blood vessels but VEGF(165) b was apical and VEGF(165) basal in RPE. Diffuse VEGF(165) immunoreactivity was prominent in 12WG innermost retina where blood vessels will form while VEGF(165) b was present in most CXCR4(+) progenitors in the inner neuroblastic layer and migrating angioblasts in the putative nerve fiber layer. By 21WG, VEGF(165) was present in nerve fibers and VEGF(165) b in the inner Muller cell process. The localization of VEGF(165) b was distinctly different from VEGF(165) both spatially and temporally and it was often associated with nucleus in progenitors., (Copyright © 2012 Wiley Periodicals, Inc.)

The pecten oculi is a highly vascular and pigmented organ placed in the vitreous body of the avian eye. As no data are currently available on the morphological organization of the pecten in the Psittaciformes, the pecten oculi of the budgerigar (Melopsittacus undulatus) was studied. The eyes from adult male budgerigars were examined by light, transmission, and scanning electron microscopy and a morphometric study on both light and transmission electron microscopy specimens was also performed in the different parts of the organ. In the budgerigar, the type of the pecten oculi was pleated. Its basal part had a cranio-caudal and postero-anterior course; its body consisted of 10-12-folds joined apically by a densely pigmented bridge. The pecten showed many capillaries, whose wall was thick and formed by pericytes and endothelial cells. These latter had a large number of microfolds, rectilinear on their luminal surface and tortuous on their abluminal surface. Interstitial pigment cells were placed among the capillaries, filled with melanin granules and showed many cytoplasmic processes. The morphometric analysis demonstrated significant differences among the three parts of the organ relative to the length of the endothelial processes and to the number and size of the pigment granules. The morphological and morphometric analysis showed that the bridge of the budgerigar, different from the other birds, had a large number of capillaries, so that this part of the organ could also play a trophic role for the retina in addition to the choriocapillaris., (Copyright © 2012 Wiley Periodicals, Inc.)

Background: Valuable insights into the complex process of retinal vascular development can be gained using models with abnormal retinal vasculature. Two such models are the recently described mouse lines with mutations in Lama1, an important component of the retinal internal limiting membrane (ILM). These mutants have a persistence of the fetal vasculature of vitreous (FVV) but lack a primary retinal vascular plexus. The present study provides a detailed analysis of astrocyte and vascular development in these Lama1 mutants., Results: Although astrocytes and blood vessels initially migrate into Lama1 mutant retinas, both traverse the peripapillary ILM into the vitreous by P3. Once in the vitreous, blood vessels anastomose with vessels of the vasa hyaloidea propria, part of the FVV, and eventually re-enter the retina where they dive to form the inner and outer retinal capillary networks. Astrocytes continue proliferating within the vitreous to form a dense mesh that resembles epiretinal membranes associated with persistent fetal vasculature and proliferative vitreoretinopathy., Conclusions: Lama1 and a fully intact ILM are required for normal retinal vascular development. Mutations in Lama1 allow developing retinal vessels to enter the vitreous where they anastomose with vessels of the hyaloid system which persist and expand. Together, these vessels branch into the retina to form fairly normal inner retinal vascular capillary plexi. The Lama1 mutants described in this report are potential models for studying the human conditions persistent fetal vasculature and proliferative vitreoretinopathy.

Purpose: To study vascular endothelial growth factor (VEGF) regulation in the development of intravitreous neovascularization and peripheral avascular retina in peripheral severe retinopathy of prematurity (ROP)., Methods: The rat 50/10 model of ROP mimics zone II, stage 3 severe ROP and recreates fluctuations in transcutaneous oxygen levels in preterm infants. On postnatal (p) day ages p0, p8, p11-p14, and p18, retinas from the model or room-air (RA) age-matched pups were analyzed for mRNA of VEGF splice variants and receptors using real-time polymerase chain reaction or VEGF protein using enzyme-linked immunosorbent assay., Results: On p14, when retinas were only 70% vascularized in the model but fully vascularized in RA, VEGF₁₆₄ expression was threefold greater in the model compared to RA. On p18, intravitreous neovascularization was associated with a 5-fold increase in VEGF₁₆₄ mRNA in the model compared to RA. By analysis of variance, VEGF₁₆₄ and VEGFR2 mRNAs were up-regulated in association with increasing developmental age (P<.0001 for both comparisons) or exposure to the model compared to RA (P<.0001 and P=.0247, respectively), whereas increasing developmental age was associated only with up-regulated VEGF₁₂₀ (P=.0006), VEGF₁₈₈ (P=.0256), and VEGFR1 (P<.0001) mRNAs. VEGF protein increased significantly in the model and on p14 and p18 compared to RA (P<.0001)., Conclusions: The model mimics contemporary severe ROP in the United States unlike other models of oxygen-induced retinopathy. Compared to RA retinas, VEGF significantly increased in association with avascular retina and intravitreous neovascularization. A hypothesis is proposed that VEGF up-regulation plays a role in the development of both important features.

Although a blockade of acetylcholine esterase has been reported to suppress neuronal cell death induced by exogenous glutamate and beta-amyloid, information is still limited regarding the neuroprotective effects of the acetylcholine esterase inhibitor donepezil. We histologically examined the effects of donepezil on neuronal injury induced by ischemia-reperfusion. Intravenous and intravitreous treatment with donepezil 15 min prior to ischemia dramatically reduced the retinal damage. The protective effect of donepezil in the ganglion cell layer was not affected by mecamylamine, a nicotinic acetylcholine-receptor antagonist, nor scopolamine, a muscarinic acetylcholine-receptor antagonist. The protective effect of donepezil in the inner plexiform layer was reduced not by mecamylamine, but by scopolamine. Neostigmine, a choline-esterase inhibitor, and pilocarpine, a muscarinic acetylcholine-receptor agonist, have protective effects in the inner plexiform layer and the inner nuclear layer. These results suggest that not only the activation of acetylcholine receptors but also a mechanism unrelated to acetylcholine-esterase inhibition contribute to the protective effect of donepezil on the ganglion cells in the ischemic-reperfused rat retina. Donepezil may be useful as a therapeutic drug against retinal diseases that cause neuronal cell death such as glaucoma with high intraocular pressure.

To determine the effect of a vascular endothelial growth factor receptor 2 tyrosine kinase (VEGFR2) inhibitor on intravitreous neovascularization (IVNV), endothelial tip cell filopodia, and intraretinal vascularization in a rat model of retinopathy of prematurity (ROP). Within 4h of birth, newborn Sprague-Dawley rat pups and their mothers were cycled between 50% and 10% oxygen daily until postnatal day (p)12. Pups were given intravitreous injections of VEGFR2 inhibitor, SU5416, or control (dimethyl sulfoxide, DMSO) and returned to oxygen cycling until p14, then placed into room air. Intravitreous neovascularization (IVNV), avascular/total retinal areas, and endothelial tip cell filopodial number and length were determined in lectin-labeled neurosensory retinal flat mounts. Cryosections or fresh tissue were analyzed for phospho-VEGFR1, phospho-VEGFR2, activated caspase-3, or phospho-beta3 integrin. Human umbilical venous (HUVECs) and human choroidal endothelial cells (ECs) were treated with VEGFR2 inhibitor to determine effect on VEGFR2 phosphorylation and on directed EC migration toward a VEGF gradient. Filopodial length and number of migrated ECs were also measured. Compared to control, the VEGFR2 inhibitor reduced VEGFR2 phosphorylation in HUVECs in vitro and clock hours and areas of IVNV but not percent avascular retina in vivo. Filopodial length and number of filopodia/EC tip cell were reduced in retinal flat mounts at doses that inhibited IVNV, whereas at lower doses, only a reduction in filopodial length/EC tip cell was found. There was no difference in phosphorylated beta3 integrin and cleaved caspase-3 labeling in VEGFR2 inhibitor-treated compared to control in vivo. Doses of the VEGFR2 inhibitor that reduced filopodial length and number of filopodia/migrating EC corresponded to reduced EC migration in in vitro models. VEGFR2 inhibitor reduced IVNV and filopodial number and length/EC tip cell without interfering with intraretinal vascularization. Reducing the number and length of filopodia/endothelial tip cell may reduce guidance cues for endothelial cells to migrate into the vitreous without interfering with migration into the retina toward a VEGF gradient.

The structure of adult human vitreous condensations (tractus, membranelles) corresponds to the structure of obliterated embryonal vasa hyaloidea. Some embryonal hyaloidal vessels and structures extend from the tunica vasculosa lentis and attach to the retina between the equator and ora serrata. In normal fetal development, these hyaloidal vessels and structures successively disappear because of apoptosis. The lower the birth weight of a premature infant, the higher the risk of retinopathy of prematurity (ROP) due to the disproportion between the regression of vasa hyaloidea (hyaloideoretinal adherences) and the proliferation of retinal vessels. This may be caused by an enhanced growth of retinal vessels (or a delayed regression of vasa hyaloidea). The growth of retinal vessels stops at a barrier formed by the retina-attaching hyaloidal remnants (vessels), resulting in a demarcation line and ROP. The same retinal vessels may then, as a fibrovascular tissue, overgrow the remnants of retina-attaching hyaloidal structures (as guiding formations) through the vitreous cavity up to the tunica vasculosa lentis. The process may result in the progression from ROP stage 2 to ROP stage 5., ((c) 2008 S. Karger AG, Basel.)

Purpose: To characterize developmental defects and the time course of Norrie disease in retinal and hyaloid vasculature during retinal development and to identify underlying molecular angiogenic pathways that may be affected in Norrie disease, exudative vitreoretinopathy, retinopathy of prematurity, and Coats' disease., Methods: Norrie disease pseudoglioma homologue (Ndph)-knockout mice were studied during retinal development at early postnatal (p) stages (p5, p10, p15, and p21). Histologic techniques, quantitative RT-PCR, ELISA, and Western blot analyses provided molecular data, and scanning laser ophthalmoscopy (SLO) angiography and electroretinography (ERG) were used to obtain in vivo data., Results: The data showed that regression of the hyaloid vasculature of Ndph-knockout mice occurred but was drastically delayed. The development of the superficial retinal vasculature was strongly delayed, whereas the deep retinal vasculature did not form because of the blockage of vessel outgrowth into the deep retinal layers. Subsequently, microaneurysm-like lesions formed. Several angiogenic factors were differentially transcribed during retinal development. Increased levels of hypoxia inducible factor-1alpha (HIF1alpha) and VEGFA, as well as a characteristic ERG pattern, confirmed hypoxic conditions in the inner retina of the Ndph-knockout mouse., Conclusions: These data provide evidence for a crucial role of Norrin in hyaloid vessel regression and in sprouting angiogenesis during retinal vascular development, especially in the development of the deep retinal capillary networks. They also suggest an early and a late phase of Norrie disease and may provide an explanation for similar phenotypic features of allelic retinal diseases in mice and patients as secondary consequences of pathologic hypoxia.

Type XVIII collagen is important in the early phase of retinal vascular development and for the regression of the primary vasculature in the vitreous body after birth. We show here that the retina in Col18a1-/- mice becomes densely vascularized by anomalous anastomoses from the persistent hyaloid vasculature by day 10 after birth. In situ hybridizations revealed normal VEGF mRNA expression, but the phenotype of collagen XVIII deficient mice closely resembled that of mice expressing VEGF120 and VEGF188 isoforms only, suggesting that type XVIII collagen may be involved in VEGF function. Type XVIII collagen was found to be indispensable for angiogenesis in the eye, as also oxygen-induced neovascularization was less intense than normal in the Col18a1-/- mice. We observed a marked increase in the amount of retinal astrocytes in the Col18a1-/- mice. Whereas the retinal vessels of wild-type mice are covered by astrocytes and the regressing, thin hyaloid vessels are devoid of astrocytes, the retinal vessels in the Col18a1-/- mice were similarly covered by astrocytes but not the persistent hyaloid vessels in the vitreous body. Interestingly, double null mice lacking type XVIII collagen and its homologue type XV collagen had the persistent hyaloid vessels covered by astrocytes, including the parts located in the vitreous body. We thus hypothesize that type XV collagen is a regulator of glial cell recruitment around vessels and that type XVIII collagen regulates their proliferation.

Purpose: To investigate the nature and origin of the vitreous membranes in mice with knock-out of the Norrie gene product (ND mice)., Methods: Eighty-two eyes of ND mice of different age groups (postnatal day [P]0-13 months) and 95 age-matched wild-type control mice were investigated. In vitreoretinal wholemounts and in sagittal sections, vessels and free cells were visualized by labeling for lectin. In addition, staining with a marker for macrophages (F4/80) and collagen XVIII/endostatin known to be involved in regression of hyaloid vessels was performed for light and electron microscopic investigations. Endostatin expression was confirmed by Western blot analysis., Results: Wild-type controls showed the typical pattern of hyaloid vessels, their regression and concomitantly retinal vasculogenesis and angiogenesis. Hyaloid vessels all stained for endostatin, whereas retinal vessels remained unstained. In ND mice, 1 to 5 days after birth, the hyaloid and retinal vasculatures were comparable to that in control mice. The hyaloid vessels also stained for endostatin. Numerous F4/80-positive cells were present adjacent to the vessels. With increasing age, only a few connecting branches of the hyaloid vessels regressed. Even in old mice most of the hyaloid vessels persisted. The vessels still stained for endostatin. Retinal angiogenesis was impaired., Conclusions: Retrolental membranes in ND mice consist of persistent hyaloid vessels, indicating that the ND gene product is important for the process of regression of these vessels. The ND gene product neither influences endostatin expression nor the presence of macrophages.

Purpose: Vascular endothelial growth factor (VEGF)-A and placental growth factor (PlGF) are members of a large group of homologous peptides identified as the VEGF family. Although VEGF-A is known to act as a potent angiogenic peptide in the retina, the vasoactive function of PlGF in this tissue is less well defined. This study has sought to elucidate the expression patterns and modulatory role of these growth factors during retinal vascular development and hyaloid regression in the neonatal mouse., Methods: C57BL6J mice were killed at postnatal days (P)1, P3, P5, P7, P9, and P11. The eyes were enucleated and processed for in situ hybridization and immunocytochemistry and the retinas extracted for total protein or RNA. Separate groups of neonatal mice were also injected intraperitoneally daily from P2 through P9 with either VEGF-neutralizing antibody, PlGF-neutralizing antibody, isotype immunoglobulin (Ig)-G, or phosphate-buffered saline (PBS). The mice were then perfused with fluorescein isothiocyanate (FITC)-dextran, and the eyes were subsequently embedded in paraffin wax or flat mounted., Results: Quantitative (real-time) reverse transcription-polymerase chain reaction (RT-PCR) demonstrated similar expression patterns of VEGF-A and PlGF mRNA during neonatal retinal development, although the fluctuation between time periods was greater overall for VEGF-A. The localization of VEGF-A and PlGF in the retina, as revealed by in situ hybridization and immunohistochemistry, was also similar. Neutralization of VEGF-A caused a significant reduction in the hyaloid and retinal vasculature, whereas PlGF antibody treatment caused a marked persistence of the hyaloid without significantly affecting retinal vascular development., Conclusions: Although having similar expression patterns in the retina, these growth factors appear to have distinct modulatory influences during normal retinal vascular development and hyaloid regression.

Intravitreal neovascular diseases are a major cause of blindness worldwide. It remains unclear why neovessels in many retinal diseases spread into the physiologically nonvascularized vitreous rather than into the ischemic retinal areas, where the angiogenic factors are released. Here we show that inducible nitric oxide synthase (iNOS) is expressed in the ischemic retina. Using iNOS knockout mice and the iNOS inhibitor 1400W, we demonstrate that iNOS expression inhibits angiogenesis locally in the avascular retina, mediated at least in part by a downregulation of VEGF receptor 2 (VEGFR2) in cells adjacent to iNOS-expressing cells. At the same time, pathological intravitreal neovascularization is considerably stronger in iNOS-expressing animals. These findings demonstrate that iNOS plays a crucial role in retinal neovascular disease and show that it offers an ideal target for the control of vitreal neovascularization through improvement of the vascularization of the hypoxic retina.

The hyaloid artery is a vestigial vessel situated on the optic nerve extending to the posterior surface of the lens in the vitreous cavity of the eye. We studied the nature and pattern of cell death during regression of the hyaloid artery. The cells comprising the hyaloid artery appear to be alive for 20 days after birth in the rat, and cell death during regression of the hyaloid artery has the characteristics of apoptosis. We observed apoptotic bodies containing condensed chromatin and identified the hyaloid vessels as targets of macrophage-mediated remodelling. Using the "TUNEL method" for labeling fragmented DNA in vascular cells, we assessed the pattern of apoptotic cell death during hyaloid artery regression. Our study demonstrated the appearance of apoptosis in pericytes as well as endothelial cells during regression in the vasculature. In pericytes, apoptosis appeared to begin or to occur more frequently than in endothelial cells. Both morphological and TUNEL analyses indicated that capillary apoptosis occurs mainly from day 10 to day 20 after birth rather than from the 1st day. Macrophages were present near the hyaloid artery and these may influence apoptosis.

Coats' disease is an idiopathic disorder in which telangiectatic and aneurysmal retinal vessels leak a lipoproteinaceous exudate, with consequent bullous retinal detachment. It is a diagnostic challenge, and CT and MR imaging provide valuable information to differentiate it from other pathologies, particularly from retinoblastoma. Typical, advanced Coats' disease shows on CT a denser substance posterior to the vitreous, which on MR is hyperintense on all pulse sequences. Contrast administration on both CT and MR might give slight linear enhancement at the boundary between vitreous and exudation. Persistent hyperplastic primary vitreous (PHPV) is a unilateral disorder in a microphthalmic eye, seen in full-term infants. PHPV rarely is bilateral in patients with Norrie's disease, Warburg syndrome, or patients with retinal dysplasia. Persistent fetal vasculature leads to fibrosis, resulting in elongation of the ciliary processes, retinal detachment, and spontaneous cataracts. The CT appearance in the disorder is quite variable; however, MR imaging may be superior in demonstrating the enhancing retrolental mass and the elongated ciliary processes.

The relation between extension of extraretinal vasoproliferation and vitreous fibers in stage 3 mild retinopathy of promaturity was evaluated by light and electron microscopy using 4 eyes obtained after autopsies. Seven developing human eyes from 20 to 40 weeks' gestation were used as controls. At an early stage, when vascular tissues broke the inner limiting membrane of the retina and grew into the vitreous cavity, they extended vertically from the retina. Proliferating extraretinal vessels extended towards the posterior surface of the lens. They ran side by side with the framework of vitreous fibers. Electron microscopy demonstrated multiple attachments between vascular endothelia and vitreous fibers. Normal developing vitreous fibers were very dense and homogeneous, running towards the lens not only from the vitreous base but also from the retina at the equator and at the posterior pole. These findings suggest that extension of extraretinal vasoproliferation in retinopathy of prematurity is induced by the framework of the developing vitreous.

The relationship of the vasculature to the neuronal layers was studied in whole-mounts and in sections of macaque retinas. Like other central nervous structures, primate retinas have local variations in vascularity that reflect local variations in metabolism, rather than simply tissue thickness or volume. A special feature of the retina is a dense vascular plexus in the nerve fiber layer, which is unmyelinated and hence must generate a substantial metabolic demand for ion pumping. Much of the retinal vasculature is laminated and located at specific layer boundaries. Throughout the central retina, two planes of capillaries bracket the inner nuclear layer to form a sclerad capillary network. In some regions, especially near the fovea, a second, more vitread network brackets the ganglion cell layer with another pair of capillary planes. Wherever the nerve fiber layer is thick, the vitread network becomes less planar and is multilayered. When surrounded by nerve fibers, capillaries tend to orient parallel to the fibers; when adjacent to ganglion cell bodies, the capillaries are less systematically oriented. At the border between the nerve fiber layer and the ganglion cell layer, rows of ganglion cells often interdigitate with nerve fiber bundles, resulting in local perturbations of capillary orientation. The volume of the sclerad capillary network is relatively constant at different locations, but the volume of the vitread network increases dramatically where the nerve fiber layer is thick. As a result, the vascularity of the retina is greatest in the peripapillary region near the optic disk, even though the total thickness of the peripapillary retina is comparable to the retinal thickness near the foveal crest. As many as 60-70% of the photons passing through the retina in the peripapillary region will encounter one or more capillaries before reaching a photoreceptor. Median capillary diameter increases with retinal depth from 4.5-4.7 microns in the nerve fiber layer to 5.0 microns at the sclerad border of the inner nuclear layer. Capillary diameter in the nerve fiber layer also increases near the optic disk.

The angioarchitecture of vitreal and retinal neovascularizations produced experimentally in the eyes of kittens aged 2 to 9 weeks was studied with scanning electron microscopy. Various forms of new retinal and vitreal vessels were observed depending on topographic locations. Intraretinal neovascularization was observed at the retinal periphery as it grew toward the avascular zone in forms of short vascular buds, aneurysmal outgrowths, and neovascular loops. Posterior or to this frond of neovascularization, intertwining intraretinal telangiectasia was observed. At the posterior pole, capillaries with microaneurysms extended posteriorly toward the deeper layers of the retina from the vascular trunks at the nerve fiber layer. Vitreal neovascularization broke through the internal limiting membrane and exhibited aneurysmal outgrowths, clusters of glomerular swellings, and sinusoidal vascular channels. At the optic disc, vitreous neovascularization took the form of aneurysmal outgrowths and long vascular buds. Vitreal neovascularization showed different characteristics from the intraretinal neovascularization. We hypothesize that the topographic variation of the angioarchitecture of retinal and vitreal neovascularizations depends on the maturity of the vessels and might be related to the hemodynamics at each site.

Presented is the personal material comprising cases treated surgically for primary retinal detachment; analysed is the causal connection between postoperatively occurring proliferative vitreoretinopathy and the risk factors. The comparison of personal results with results of the other authors shows that in case of diagnosis of risk factors the reduction of the surgical trauma reduces the threat of proliferative vitreoretinopathy after operation of retinal detachment.

Analysed were the results of surgical treatment, causes of the failure and early recurrence in 108 patients with retinal detachment in whom was performed an indentation of the sclera by means of a balloon (1st group--50) or by an episcleral implant (2d group--58). In the first group the apposition was achieved in 92 p.c., in the 2d group in 90 p.c. An early recurrence (2 weeks to 2 months) was observed in 9 eyes which were operated again achieving an apposition of the retina in 6 eyes. There were following causes of the failure: the existence of a second hole, rupture of the balloon, PVR before surgery and traction of the retina in the region of the hole. An early recurrence occurred in the consequence of intensification of the PVR together with inaccurate localization of the implant, existence of the PVR before the operation and traction of the retina in the vicinity of the hole, formation of a new hole.

A method is presented that demonstrates the whole network of the blood capillaries of the human retina, making it possible (using a dark-field microscope at high magnification) to view the three-dimensional arrangement of this network in different planes by moving the drive of the microscope. The basic principle includes filling the vascular system with oxygen in statu nascendi by injection of H2O2 into the vitreous body of the intact eye and achieving fixation of the retinal tissue by application of a mixture of H2O2 and ethanol, and rendering the dry and flat retinal preparation transparent by soaking it in a specially prepared resin. The retinal vessels are made visible in an optically empty dark field by diffraction of light in a gas-filled specimen. This method allows preparation of the human retinal vasculature in simply enucleated eyes without cannulation of the central retinal artery and, therefore, it may be appropriate to support pathological and anatomical investigations.

There is a bimodal distribution of retinal thicknesses among mammals. Avascular retinae that are totally dependent on the choroid measure less than the theoretical oxygen diffusion maximum of 143 microns. Vascular retinae often measure twice this distance, but their avascular portions do not exceed the calculated diffusion maximum. Avascular retinae also show heavy glycogen deposition in the Müller cells of the inner retina, short photoreceptors, and lack both a tapetum and retinal taper. It is argued (1) that these anatomical features represent adaptations to their nutritional dependence on the choroid, and (2) that the phylogenetic distribution of avascularity (monotremes, many marsupials, and some of the primitive placental mammals) suggests that it is the ancestral condition in mammals. Finally, it is noted that some of the pathologic changes associated with retinal vascular damage in humans, namely thinning, gliosis, and glycogen deposition, are reminiscent of these ancestral patterns for coping with a limited blood supply.

A 50-year-old man, diagnosed as having familial primary amyloidosis due to his systemic symptoms, family history, and to histological examination of biopsy specimens, was examined by fluorescein angiography. The following results were obtained: (1) The sheathing retinal artery showed a defect of dye filling in one region, but not in the other region. Dye leakage from the sheathing artery was observed. (2) The configuration of macular vessels was rough, a microaneurysma-like change and the leakage of dye were also recognized in the capillaries of this region. (3) It was speculated that vitreous opacity originated from vascular lesions.

Preretinal vascular loops are a congenital malformation usually occurring in otherwise normal fundi. These anomalies consist of vessels, most commonly arteries, that spiral into the vitreous and return to the otherwise normal retinal vascular system. The authors present two members of a family with preretinal vascular loops and two members with other vascular anomalies. The inheritance pattern in this group is autosomal dominant.

Ten choriovitreal and three subretinal fibrovascular proliferations were seen after heavy focal photocoagulation of diabetic retinopathy. These complications were believed to be caused by the use of a small spot size with high energy.

Two young patients (27 and 30 years old, respectively) with long-standing diabetes mellitus underwent vitreoretinal surgery for traction retinal detachment and vitreous hemorrhage. The postoperative course in each was characterized by early recurrence of the hemorrhage into the vitreous cavity, hypotony, and severe visual loss. Histopathologic examination of both operated on eyes showed anterior extraretinal fibrovascular proliferation extending along the anterior hyaloid to the posterior lens surface, causing traction detachments of the peripheral retina and ciliary body. The vessels originated from the anterior retina. There was no evidence of excess fibrous tissue at the well-healed sclerotomy wounds.

Different stages of retino-choroiditis, many of toxoplasmic origin, may develop vascular alterations. One of these pictures has been observed in relation to old scars of retino-choroiditis, where focal vasculitis may affect the venous return circulation sometimes leading to intraocular hemorrhages and retinal holes of variable severity. The natural history of this process is described, as well as the clinical characteristics and the results of treatment. Generally, the disease has a favorable course and a good prognosis. 18 cases are reported.

The blood-retinal barrier permeability to fluorescein was determined in 20 eyes from 17 normal volunteers (mean age 31 years) and in 20 eyes from 19 juvenile diabetics without apparent retinopathy (mean age 35 years - mean duration of diabetes 6 years). The permeability was in normal subjects (1.1 +/- 0.4) X 10(-7) cm/sec (mean +/- 2 X SD) and in juvenile diabetics (1.1 +/- 0.7) X 10(-7) cm/sec (mean +/- 2 X SD). Thus a break-down of the blood-retinal barrier cannot be demonstrated as a very early and general phenomenon in the early course of the diabetic disease. The fluorescein diffusion coefficient in the vitreous body was determined and juvenile diabetics without apparent retinopathy showed a diffusion coefficient of (0.80 +/- 0.25) X 10(-5) cm2/sec (mean +/- 2 X SD), which was the same as in normals where the diffusion coefficient was (0.69 +/- 0.46) X 10(-5) cm2/sec (mean +/- 2 X SD).

The glycogen content in the individual eye tissues is strongly correlated to blood supply. Our investigations on the retina of bovines, which have not been fully evaluated, show that the time interval between interruption of blood supply and preparation of the retina is of special importance. Pressure ischemia affects a decrease in glycogen content in the retina and vitreous of rabbits, which is, however, less distinct in the vitreous. Decrease of glycogen with ischemia also takes place in the cornea and, to a lesser degree, in iris and choroid. In contrast, there is no decrease in the glycogen content of the lens. Changes in glycogen content of the rabbit retina after ligation of the A. carotis communis is less distinct than with pressure ischemia. In the vitreous, changes in glycogen content could not be observed. Values measured in both tissues of the ligated eye decrease with additional pressure ischemia.

Forty-two eyes in 30 patients with disc neovascularization were treated by the method of extensive photocoagulation of the retinal periphery (ablation). Total obliteration of disc neovascularization was accomplished in 76% of eyes treated. Partial regression was noted in 21% of eyes treated. There were only 2 major complications--traction retinal detachment and one major hemorrhage--in this series. Feeder-frond treatment was necessary to obliterate recurrent disc neovascularization in 2 eyes. It is concluded that therapy is efficacious in the treatment of papillary neovascularization.

In cases with persistent primary vitreous in full-term infants there were vitreoretinal reactions including vitreoretinal anastomoses of blood vessels, vitreoretinal strands, traction folds of the retina, avascular retinal areas, hypoplasia of retinal blood vessels, rarefaction of retinal ganglion cells, and ischaemic structural changes of the retina. New vessel formation on the surface of avascular retinal areas was supplied from persistent hyaloid blood vessels. This finding may be of some importance in explaining those cases histologically resembling classical retrolental fibroplasia with no history of prematurity or oxygen treatment.

Two patients with talc emboli of the retinal vasculature, both of them intravenous drug abusers, are described. One underwent a pars plana vitrectomy for removal of the vascularized vitreous mass that obscured the retina. The vitreous mass was presumably a result of retinal neovascularization secondary to the talc embolic. Other ocular findings that may be observed in the intravenous drug abuser are described.

The retinae of two species of teleosts, the rippled blenny Istiblennius edentulus and the blue tusk fish Choerodon albigena are examined in wholemount. The retinal topography of Nissl-stained cells within the ganglion cell layer in each species reveals a temporal (4.32 x 10(4) cells per mm2) and a nasal (3.83 x 10(4) cells per mm2) area centralis in the rippled blenny and two temporal areae centrales (8.30 x 10(4) and 8.00 x 10(4) cells per mm2) and a horizontal streak (5.00 x 10(4) cells per mm2) in the tusk fish. These areas are thought to subserve higher spatial resolution. Transcardial perfusions of indian ink reveal an extensive network of vitreal blood vessels which are supplied by the hyaloid artery and overlie the retina in each species. This rich network of vitreal vessels supplies areas of increased ganglion cell density although areas of maximum cell density are devoid of vessels to preserve the high spatial resolving power of the eye within this region. Unique blood vessel plexuses overlying the optic disc and falciform process in the tusk fish are also described. The diameter of the overlying vitreal vessels is compared to the soma sizes of cells within the ganglion cell layer.

A patient with proliferative diabetic retinopathy was treated by panretinal and focal photocoagulation. Later, he developed one area of clinically diagnosed chorioretinal and choriovitreal neovascularization (CNV), neovascular glaucoma, and a blind painful eye necessitating enucleation. Clinicopathologic correlations of this eye including fundus photography, fluorescein angiography, light and electron microscopy are reported. Histopathologic examination revealed three areas of CNV, suggesting that some CNV may go undetected clinically also in other cases and thus may occur more frequently than evident from the literature. Our CNV occurred at sites of focal treatment. Retreatment of one area was unsuccessful. Choriovitreal neovascularization passed through discontinuities of Bruch's membrane into the retina and showed fenestrae of the endothelial cells. Endothelial fenestrae may account for the profuse fluorescein leakage seen clinically in CNV.