Pigment epithelium-derived factor: a potent inhibitor of angiogenesis.
(33/2152)
In the absence of disease, the vasculature of the mammalian eye is quiescent, in part because of the action of angiogenic inhibitors that prevent vessels from invading the cornea and vitreous. Here, an inhibitor responsible for the avascularity of these ocular compartments is identified as pigment epithelium-derived factor (PEDF), a protein previously shown to have neurotrophic activity. The amount of inhibitory PEDF produced by retinal cells was positively correlated with oxygen concentrations, suggesting that its loss plays a permissive role in ischemia-driven retinal neovascularization. These results suggest that PEDF may be of therapeutic use, especially in retinopathies where pathological neovascularization compromises vision and leads to blindness. (+info)
Automated localisation of the optic disc, fovea, and retinal blood vessels from digital colour fundus images.
(34/2152)
AIM: To recognise automatically the main components of the fundus on digital colour images. METHODS: The main features of a fundus retinal image were defined as the optic disc, fovea, and blood vessels. Methods are described for their automatic recognition and location. 112 retinal images were preprocessed via adaptive, local, contrast enhancement. The optic discs were located by identifying the area with the highest variation in intensity of adjacent pixels. Blood vessels were identified by means of a multilayer perceptron neural net, for which the inputs were derived from a principal component analysis (PCA) of the image and edge detection of the first component of PCA. The foveas were identified using matching correlation together with characteristics typical of a fovea-for example, darkest area in the neighbourhood of the optic disc. The main components of the image were identified by an experienced ophthalmologist for comparison with computerised methods. RESULTS: The sensitivity and specificity of the recognition of each retinal main component was as follows: 99.1% and 99.1% for the optic disc; 83.3% and 91.0% for blood vessels; 80.4% and 99.1% for the fovea. CONCLUSIONS: In this study the optic disc, blood vessels, and fovea were accurately detected. The identification of the normal components of the retinal image will aid the future detection of diseases in these regions. In diabetic retinopathy, for example, an image could be analysed for retinopathy with reference to sight threatening complications such as disc neovascularisation, vascular changes, or foveal exudation. (+info)
Peripapillary retinal blood flow in normal tension glaucoma.
(35/2152)
AIMS: To determine if normal tension glaucoma (NTG) patients differ from age matched controls in blood flow to the peripapillary retina, as measured with confocal scanning laser Doppler flowmetry (cSLDF; "Heidelberg retinal flowmetry"). METHODS: 12 NTG patients and 12 age matched controls were compared using (a) 10 x 10 pixel boxes (the instrument default sample size), taken from the nasal and temporal peripapillary retina, (b) the average from two of these boxes, and (c) every qualifying pixel within the peripapillary retina. RESULTS: Patients and controls did not differ in blood flow measured using the default sample from a single 10 x 10 pixel box, placed in either the temporal or nasal peripapillary retina, or expressed as the average from these two boxes. However, in histograms using every pixel from the peripapillary retina, NTG patients displayed significantly higher percentages of minimal flow pixels (defined as less than one arbitrary unit of flow: 30% v 19%, p < 0.01), and significantly lower flow in the 25th, 50th, and 75th percentile flow pixel (each p < 0.05) than did age matched controls. CONCLUSION: NTG is characterised by reduced blood flow in the peripapillary retina, a result suggesting that blood flow deficits accompany, and perhaps may contribute to, disease development in these patients. (+info)
Intraretinal oxygen distribution in the rat with graded systemic hyperoxia and hypercapnia.
(36/2152)
PURPOSE: To describe the nature of oxygen level changes in specific layers in the rat retina under graded levels of systemic hyperoxia, with and without hypercapnia. METHODS: Oxygen-sensitive microelectrodes were used to measure oxygen tension as a function of depth through the retina of anesthetized, mechanically ventilated rats. Breathing mixtures were manipulated to produce stepwise increments in systemic oxygen levels, with or without 5% CO2. Retinal arteriovenous oxygen differences were also measured as an indicator of oxygen delivery through the retinal circulation. Systemic blood gas levels were measured under each condition. RESULTS: Hyperoxia increases PO2 throughout the retina to a varying extent in different retinal layers, with the increase more pronounced in the outer retina than in the inner retina. Simultaneous hypercapnia results in further increases in retinal oxygen levels. The lowest intraretinal oxygen level was consistently found in the inner plexiform layer (IPL), between the two capillary layers that support this region. There was a greater than fourfold increase in oxygen supply from the choroid with hyperoxia but, remarkably, the retinal circulation continued to provide a net delivery of oxygen to the retina. CONCLUSIONS: Hyperoxia results in a significant but nonuniform increase in oxygen level in all layers of the rat retina, which is augmented by hypercapnia. The persistence of a minimum oxygen level in the IPL, despite the dramatic increase in oxygen flux from the choroid, suggests that oxygen consumption increases significantly in the IPL under hyperoxic conditions. (+info)
17 Beta-estradiol increases VEGF receptor-2 and promotes DNA synthesis in retinal microvascular endothelial cells.
(37/2152)
PURPOSE: Estrogen is known to promote angiogenesis in gonads. The presence of estrogen receptors in the vascular endothelium of organs other than gonads has been reported. The goal of this study was to determine whether estrogen promotes the proliferation of retinal microvascular endothelial cells and to explore the mechanism of it. METHODS: DNA was quantitated using primary cultures of bovine retinal endothelial cells that were incubated with different doses of 17 beta-estradiol (E2), VEGF, or both. The changes in expression level of VEGF and VEGF receptor-2 (VEGFR2) were measured using northern blot analysis after treatment with E2. The presence of estrogen receptors in the endothelial cells was studied by immunohistochemistry and western blot analysis. RESULTS: 17 Beta-estradiol (E2) increased the DNA level in bovine retinal capillary endothelial cells (BRECs) by 177% at 1 nM (P < 0.05) and 150% at 10 nM (P < 0.05) by comparison with unstimulated BREC. One hundred nanomole tamoxifen completely blocked the E2-induced DNA synthesis in BRECs. Ten nanomole E2 augmented vascular endothelial growth factor (VEGF)-induced DNA synthesis in BRECs significantly (160%, P < 0.01). Ten nanomole E2 also increased VEGF mRNA expression, which peaked after 24 hours (6.7 times, P < 0.05), and VEGF receptor-2 (VEGFR2) mRNA expression, which peaked after 9 hours (2.4 times, P < 0.05). The mRNA expression level of VEGFR2 peaked with 10 nM E2 (P < 0.05) and that of VEGF reached maximum with 1 nM E2 (15 times, P < 0.001). VEGFR2 and VEGF proteins increased in parallel with their mRNA levels. Immunocytochemistry showed estrogen receptor expression in BRECs, and western blot analysis indicated the presence of a 67-kDa protein that was compatible with the estrogen receptor. CONCLUSIONS: These findings suggest that E2 may stimulate BREC growth by the receptor-mediated pathway and that E2 may augment the VEGF-dependent angiogenesis partly through the upregulation of VEGFR2. (+info)
Administration of nerve growth factor, brain-derived neurotrophic factor and insulin-like growth factor-II protects phosphate-activated glutaminase in the ischemic and reperfused rat retinas.
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Phosphate-activated glutaminase (PAG) activity decreases markedly in the early period of ischemia. The decrease of the enzyme activity is reversible if the ischemic period is relatively short, but it becomes irreversible after 90 minutes of ischemia. The deterioration is a functional damage of the retinas caused by ischemia. We studied effects of growth factors and neurotrophic factors on protection of PAG in the ischemic and reperfused rat retinas. Before ischemia, 1 microl of growth factors or neurotrophic factors (0.1 microg/microl for insulin-like growth factor-I [IGF-I], insulin-like growth factor-II [IGF-II], brain-derived neurotrophic factor [BDNF], nerve growth factor [NGF]; 1 microg/microl for basic fibroblast growth factor [bFGF]) were injected into the vitreous cavity of the left eyes of anesthetized Sprague Dawley rats. As a control, phosphate buffered saline was injected to the right eyes. To induce ischemia, we clamped left eyes for 90 minutes after bulbar conjunctival incision all around limbus. The rat retinas were homogenized with distilled water 1 day after reperfusion and used for PAG assay. Retinal ammonia concentration was also determined as a ischemic marker. About 80% decrease of retinal PAG activity and 50% increase of retinal ammonia concentration were observed after 90 minutes of ischemia and 1 day of reperfusion as compared with unoperated normal eyes. IGF-II, BDNF and NGF had protective effects on the retinal PAG activity, whereas IGF-I, bFGF, stable bFGF were less effective. In addition, IGF-II and BDNF suppressed elevation of retinal ammonia concentration. BDNF, NGF and IGF-II have marked effect on the protection of PAG activity in the ischemic and reperfused rat retinas, whereas bFGF, which is very effective for the protection of ischemic cell death, shows moderate effect. (+info)
Light and choroidal PO2 modulation of intraretinal oxygen levels in an avascular retina.
(39/2152)
PURPOSE: To determine the influence that choroidal oxygen level and outer retinal oxygen demand have on oxygen availability to the inner retina when the choroid is the only source of retinal oxygenation. This condition prevails in avascular retinas and in vascularized retinas suffering vascular occlusion. METHODS: Oxygen-sensitive microelectrodes were used to measure the oxygen tension as a function of depth in the naturally avascular retina of anesthetized and mechanically ventilated guinea pigs (n = 6). Choroidal PO2 was manipulated by varying the ventilation gas mixture, and outer retinal oxygen consumption was modulated by light-dark adaptation. Individual PO2 profiles were fitted to a multilayer mathematical model of PO2 distribution, and pairs of profiles at different choroidal PO2 levels, or under light and dark conditions, were fitted to an intraretinal PO2 difference model. Both models reflect the purely choroidal supply of retinal oxygenation. RESULTS: An increase in choroidal PO2 produced an equivalent increase in all retinal layers. Light induced a decreased oxygen consumption in the region of the inner segments of the photoreceptors, which resulted in a significant increase in PO2 in this layer, flowing on unattenuated to all inner retinal layers. The intraretinal PO2 distribution and the light- and ventilatory-induced changes in PO2 were consistent with theoretical predictions of the mathematical models. CONCLUSIONS: The present experimental studies confirm that when the choroid is the only source of retinal oxygenation, the full effect of increased choroidal oxygen level or reduced uptake in the outer retina passes through to the inner retinal layers if the oxygen utilization by the inner retina remains constant. (+info)
Remodeling of retinal capillaries in the diabetic hypertensive rat.
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PURPOSE: To document the effect of sustained systemic hypertension on the integrity and ultrastructural morphology of retinal capillaries in diabetic and nondiabetic rats. METHODS: Normotensive (strain Wistar-Kyoto; WKY) and genetically hypertensive (spontaneously hypertensive; SHR) rats were rendered diabetic by intravenous streptozotocin injection. At 20 weeks of diabetes, endothelial cells, pericytes, and extracellular matrix were evaluated by ultrastructural morphometry. Serum albumin was localized by immunofluorescence microscopy. RESULTS: The endothelial cell layer was markedly thinner in the diabetic normotensive animals. The number of intercellular junctions was reduced in both the nondiabetic and diabetic hypertensive group but less so in the diabetic normotensive group. No significant endothelial cell loss was noted in either of the experimental groups, whereas the number of pericytes and the number of their cytoplasmic processes were reduced in diabetic and hypertensive animals. Significant thickening of the basement membrane and increased permeability to serum albumin were observed in diabetic and hypertensive rats and were strongly enhanced in the combined diseases. CONCLUSIONS: Endothelial thinning and shape changes from an elaborate to a simpler form as well as rounding up of the pericytes and loosening of their vascular sheaths indicate remodeling of the vascular wall during chronic diabetes and sustained hypertension, before a characteristic vasculopathy becomes manifest. The combination of diabetes and hypertension enhances these features, as well as basement membrane thickening and breakdown of the blood-retinal barrier. (+info)