Three-dimensional Imaging Coupled with Topological Quantification Uncovers Retinal Vascular Plexuses Undergoing Obliteration
Chih‐Chiang ChangAlison ChuScott A. MeyerYichen DingMichel M. SunParinaz AbiriKyung In BaekVarun GudapatiXili DingPierre GuihardKristina I. BoströmSong LiLynn K. GordonJie ZhengTzung K. Hsiai
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Murine models provide microvascular insights into the 3-D network disarray seen in retinopathy and cardiovascular diseases. Light-sheet fluorescence microscopy (LSFM) has emerged to capture retinal vasculature in 3-D, allowing for assessment of the progression of retinopathy and the potential to screen new therapeutic targets in mice. We hereby coupled LSFM, also known as selective plane illumination microscopy, with topological quantification, to characterize the retinal vascular plexuses undergoing preferential obliteration.Keywords:
Plexus
Abstract The manner in which new cells are added to the growing adult goldfish retina was examined using 3 H‐thymidine radioutography. Cell proliferation leading to the formation of neurons is restricted to the retinal margin at the ora terminalis . New retina is added in concentric rings, with slightly more growth dorsonasally. The rate of cell addition is variable, averaging 12,000 cells/ day. These new cells account for about 20% of the total increase in retinal area; the remaining 80% is due to hypertrophy, or expansin, of the retina. In contrast to all of the other retinal cells, the rods do not appear to participate in the retinal expansion. This hypothesized immobility of the rods would create a shearing strain between the retinal layers resulting in a shift in their position relative to the other cells. Were they to maintain synaptic contacts with the same horizontal and bipolar cells, the rod axons would have to be elongated peripherally or the post‐synaptic cell dendrites displaced centrally. Since neurons with this morphology have not been found in the goldfish retina, these observations suggest that the rods must be changing their synaptic connections as the retina grows.
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Abstract The pattern of retinal vasculative is described and the position at which cell proliferation at the ventral retinal margin is maximal was shown to be at the point of entry of the ventral blood vessels. To test whether there is a causal relation between retinal blood supply and retinal cell production, surgical inversion of the eye, transplantations and excisions of retina were done to change the pattern of retinal vasculature. The growth pattern of inverted eyes was normal with respect to the internal axes of the eyes. After excision of part of the retina or after fusion of retinal fragments to form compound eyes, the pattern of retinal cell proliferation was not correlated with the distribution of retinal blood vessels, but was correlated with the position(s) of the choroidal fissure(s).
Blood supply
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To observe and estimate the distribution the changes of histology and biochemical parameters in retina of the patient with diabetic retinopathy.Retinas were divided into quadrants and the lesions of diabetic retinopathy were compared among quadrants. the vasculature was isolated by trypsin-digest, homogenates taken from temporal and nasal quadrant were used to explore regional differences in the expression of Glut-1, PKCbeta and activity of caspase1.Microaneurysms, acellular capillaries and pericytes ghosts were significantly increased in the temporal retina than in the nasal retina. Activity of the pro-inflammatory protease, caspase1, was the only biochemical abnormality which was found significantly difference between quadrants. Glut-1 expression was decreased in diabetes, but there was no significant difference between quadrants. Expression of PKCbeta in both temporal and nasal retina was tended to be greater in diabetic than in non diabetic patients.Retinal microvascular disease does not develop uniformly across the retina, even the different regions are exposed to the same level of hyperglycemia. Regional differences play an important role in the response of the retinal microvasculature to hyperglycemia.
Quadrant (abdomen)
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Expression and Significance of Macrophage Migration Inhibitory Factor in the Retina of Diabetic Rats
Objective To detect the expression of macrophage migration inhibitory factor (MIF) in the retina of diabetic rats,and to investigate its role in the pathogenesis of diabetic retinopathy(DR).Methods Diabetic rats model was set by intraperitoneal injection of streptozocin (STZ),and the normal control group was set too,n=20.After 8 weeks,the different content of MIF in retina of the two groups were detected by ELISA.HE staining and immunohistochemical staining were used to observe the different depression of MIF in retina.Results Compared with the normal control group,blood glucose levels increased,the expression of MIF in retina increased significantly in the diabetic group.The difference was statistically significant(all P0.01).Conclusion The expression of MIF in retina of diabetic rats increased,related to the retinal inflammatory change may participate in early diabetic retinopathy.
Pathogenesis
Intraperitoneal injection
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Spatial Pattern and Temporal Evolution of Retinal Oxygenation Response in Oxygen-Induced Retinopathy
To determine the spatial pattern and temporal evolution of the change in retinal partial oxygen pressure (DeltaPO(2)) associated with a murine oxygen-induced retinopathy (OIR) model of retinal neovascularization (NV).On P7, newborn C57BL/6 mice were exposed to 75% oxygen until postnatal day (P)12, followed by recovery in room air until P17 or P34. Control mice remained in room air until P17 or P34. At P17 and P34, functional magnetic resonance imaging (MRI) and a carbogen inhalation challenge was used to measure retinal DeltaPO(2). Retinal avascularity, distance from the optic nerve head to the vascular edge in the peripheral retina, and NV incidence and severity were measured in retinas stained with adenosine diphosphatase (ADPase).In P17 and P34 controls and in P34 OIR animals, retinas were fully vascularized without evidence of NV. In P17 OIR mice, there was a large central retinal capillary-free zone (22% +/- 3% of the entire retinal area, mean +/- SD) and 4 clockhours (range 1-7) of retinal NV at the border of the peripheral vascular and central acapillary retina in 100% (36/36) of the mice. In P17 OIR mice, retinal DeltaPO(2) over the vascularized far peripheral retina was not significantly (P > 0.05) different from the P17 control but was supernormal (P < 0.05) over the central capillary-free retina. However, no differences (P > 0.05) in retinal DeltaPO(2) were found between the P34 control and OIR groups.A reversible supernormal DeltaPO(2) was found only over the central acapillary retina during the appearance of retinal NV in a mouse OIR model. The present data show the applicability of carbogen-challenge functional MRI to the study of retinal DeltaPO(2) in vivo in eyes that are too small for the use of existing techniques.
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The oxygen requirements of different retinal layers are of interest in understanding the vulnerability of the retina to hypoxic damage in retinal diseases with an ischemic component. Here, we report the first measurements of retinal oxygen consumption in the visual streak of the rabbit retina, the region with the highest density of retinal neurons, and compare it with that in the less-specialized region of the retina underlying the vascularized portion of the rabbit retina. Oxygen-sensitive microelectrodes were used to measure oxygen tension as a function of retinal depth in anesthetized animals. Measurements were performed in the region of the retina containing overlying retinal vessels and in the center of the visual streak. Established mathematical analyses of the intraretinal oxygen distribution were used to quantify the rate of oxygen consumption in the inner and outer retina and the relative oxygen contributions from the choroidal and vitreal sides. Outer retinal oxygen consumption was higher in the visual streak than in the vascularized area (means ± SE, 284 ± 20 vs. 210 ± 23 nl O 2 ·min –1 ·cm –2 , P = 0.026, n = 10). However, inner retinal oxygen consumption in the visual streak was significantly lower than in the vascular area (57 ± 4.3 vs. 146 ± 12 nl O 2 ·min –1 ·cm –2 , P < 0.001). We conclude that despite the higher processing requirements of the inner retina in the visual streak, it has a significantly lower oxygen consumption rate than the inner retina underlying the retinal vasculature. This suggests that the oxygen uptake of the inner retina is regulated to a large degree by the available oxygen supply rather than the processing requirements of the inner retina alone.
Streak
Oxygen tension
Retinal Artery
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Objective Investigate and analyze the macular thickness in diabetic patients by optical coherence tomography(OCT) measurements.Methods There were 30 eyes(30 cases) in normal control group,and 90 eyes(30 cases) in diabetic group.The diabetic patients were divided into three groups: non-diabetic retinopathy(NDR),non-proliferative diabetic retinopathy(NPDR) and proliferative diabetic retinopathy(PDR).We observed the different tomographic features of diabetic macula and measured macular thickness.For each eye,nine different sectors were analyzed(central subfield,four parafoveal sectors,and four extrafoveal sectors).All the people had undergone examination of BCVA,slit lamp,ocular fundus,FFA and OCT.Results he average retinal thickness at central subfield(CSF) with 1 mm diameter respectively were NDR Group(225.8±18.0)μm,NPDR Group(246.8±34.2)μm,PDR Group(323.5±92.2)μm,Contral Group(224.8±15.9)μm.There were statistically significant differences in CSF among NPDR group,PDR group and the control eyes(P 0.05).But no significant difference between NDR group and the control eyes.Conclusion OCT can detect macular thickness and structural changes in eyes with DM.It is a useful tool to diagnose and monitor diabetic maculopathy.
Fundus (uterus)
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