Diabetic Retinopathy: Retina‐Specific Methods for Maintenance of Diabetic Rodents and Evaluation of Vascular Histopathology and Molecular Abnormalities

Veenstra, Alexander A.; Liu, Haitao; Lee, Chieh Allen; Du, Yunpeng; Tang, Jie; Kern, Timothy S.

doi:10.1002/9780470942390.mo140190

Cited by 49 publications

(85 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…22 When devoid of neuronal cells, the isolated vasculature was placed on a glass microscope slide, dried overnight, stained with hematoxylin and periodic acid Schiff, dehydrated, and coverslipped. Degenerated (acellular) capillaries were quantitated in up to six to seven field areas corresponding to the midretina (200× magnification) in a masked manner.…”

Section: Methodsmentioning

confidence: 99%

Photoreceptor Cells Influence Retinal Vascular Degeneration in Mouse Models of Retinal Degeneration and Diabetes

Liu

Tang

et al. 2016

Invest. Ophthalmol. Vis. Sci.

Self Cite

View full text Add to dashboard Cite

PurposeLoss of photoreceptor cells is associated with retinal vascular degeneration in retinitis pigmentosa, whereas the presence of photoreceptor cells is implicated in vascular degeneration in diabetic retinopathy. To investigate how both the absence and presence of photoreceptors could damage the retinal vasculature, we compared two mouse models of photoreceptor degeneration (opsin−/− and RhoP23H/P23H ) and control C57Bl/5J mice, each with and without diabetes.MethodsRetinal thickness, superoxide, expression of inflammatory proteins, ERG and optokinetic responses, leukocyte cytotoxicity, and capillary degeneration were evaluated at 1 to 10 months of age using published methods.ResultsRetinal photoreceptor cells degenerated completely in the opsin mutants by 2 to 4 months of age, and visual function subsided correspondingly. Retinal capillary degeneration was substantial while photoreceptors were still present, but slowed after the photoreceptors degenerated. Diabetes did not further exacerbate capillary degeneration in these models of photoreceptor degeneration, but did cause capillary degeneration in wild-type animals. Photoreceptor cells, however, did not degenerate in wild-type diabetic mice, presumably because the stress responses in these cells were less than in the opsin mutants. Retinal superoxide and leukocyte damage to retinal endothelium contributed to the degeneration of retinal capillaries in diabetes, and leukocyte-mediated damage was increased in both opsin mutants during photoreceptor cell degeneration.ConclusionsPhotoreceptor cells affect the integrity of the retinal microvasculature. Deterioration of retinal capillaries in opsin mutants was appreciable while photoreceptor cells were present and stressed, but was less after photoreceptors degenerated. This finding proves relevant to diabetes, where persistent stress in photoreceptors likewise contributes to capillary degeneration.

show abstract

Section: Methodsmentioning

confidence: 99%

Photoreceptor Cells Influence Retinal Vascular Degeneration in Mouse Models of Retinal Degeneration and Diabetes

Liu

Tang

et al. 2016

Invest. Ophthalmol. Vis. Sci.

Self Cite

View full text Add to dashboard Cite

show abstract

“…1a). This trypsin-induced retinal digestion and vascular network isolation technique was originally developed in 1993 [51] and subsequently modified by replacing trypsin with commercially available elastase [77]. Our modified protocol is as follows: retinal strips from human donors or mouse whole retinas preserved in PFA were first washed in lukewarm running distilled water overnight, then digested in 40 U/ ml elastase solution (Merck Millipore, Burlington, MA) for 2 h at 37 °C.…”

Section: Retinal Vascular Isolation and Immunofluorescent Stainingmentioning

confidence: 99%

“…To exclusively investigate the extent of retinal microvascular amyloidosis and possible pericyte degeneration in AD without interference from other retinal tissues, we enzymatically digested retinas, preserved solely the vascular network [51,77], and subsequently conducted fluorescent immunostaining for blood vessels (lectin), PDGFRβ, and different types of Aβ ( Fig. 1; extended data in Supplementary Fig.…”

Section: Retinal Pericyte Loss Along With Vascular Aβ Deposits Includmentioning

confidence: 99%

Identification of early pericyte loss and vascular amyloidosis in Alzheimer’s disease retina

et al. 2020

View full text Add to dashboard Cite

Pericyte loss and deficient vascular platelet-derived growth factor receptor-β (PDGFRβ) signaling are prominent features of the blood-brain barrier breakdown described in Alzheimer's disease (AD) that can predict cognitive decline yet have never been studied in the retina. Recent reports using noninvasive retinal amyloid imaging, optical coherence tomography angiography, and histological examinations support the existence of vascular-structural abnormalities and vascular amyloid β-protein (Aβ) deposits in retinas of AD patients. However, the cellular and molecular mechanisms of such retinal vascular pathology were not previously explored. Here, by modifying a method of enzymatically clearing non-vascular retinal tissue and fluorescent immunolabeling of the isolated blood vessel network, we identified substantial pericyte loss together with significant Aβ deposition in retinal microvasculature and pericytes in AD. Evaluation of postmortem retinas from a cohort of 56 human donors revealed an early and progressive decrease in vascular PDGFRβ in mild cognitive impairment (MCI) and AD compared to cognitively normal controls. Retinal PDGFRβ loss significantly associated with increased retinal vascular Aβ 40 and Aβ 42 burden. Decreased vascular LRP-1 and early apoptosis of pericytes in AD retina were also detected. Mapping of PDGFRβ and Aβ 40 levels in pre-defined retinal subregions indicated that certain geometrical and cellular layers are more susceptible to AD pathology. Further, correlations were identified between retinal vascular abnormalities and cerebral Aβ burden, cerebral amyloid angiopathy (CAA), and clinical status. Overall, the identification of pericyte and PDGFRβ loss accompanying increased vascular amyloidosis in Alzheimer's retina implies compromised blood-retinal barrier integrity and provides new targets for AD diagnosis and therapy.

show abstract

“…Trypsin digestion of the retina was performed according to a previously published protocol [32,33]. Briefly, eyeballs were enucleated and incubated in 4% paraformaldehyde overnight.…”

Section: Acellular Capillaries Quantificationmentioning

confidence: 99%

Characterizing the Retinal Phenotype in the High-Fat Diet and Western Diet Mouse Models of Prediabetes

Asare-Bediako

Noothi

Calzi

et al. 2020

Cells

View full text Add to dashboard Cite

We sought to delineate the retinal features associated with the high-fat diet (HFD) mouse, a widely used model of obesity. C57BL/6 mice were fed either a high-fat (60% fat; HFD) or low-fat (10% fat; LFD) diet for up to 12 months. The effect of HFD on body weight and insulin resistance were measured. The retina was assessed by electroretinogram (ERG), fundus photography, permeability studies, and trypsin digests for enumeration of acellular capillaries. The HFD cohort experienced hypercholesterolemia when compared to the LFD cohort, but not hyperglycemia. HFD mice developed a higher body weight (60.33 g vs. 30.17g, p < 0.0001) as well as a reduced insulin sensitivity index (9.418 vs. 62.01, p = 0.0002) compared to LFD controls. At 6 months, retinal functional testing demonstrated a reduction in a-wave and b-wave amplitudes. At 12 months, mice on HFD showed evidence of increased retinal nerve infarcts and vascular leakage, reduced vascular density, but no increase in number of acellular capillaries compared to LFD mice. In conclusion, the HFD mouse is a useful model for examining the effect of prediabetes and hypercholesterolemia on the retina. The HFD-induced changes appear to occur slower than those observed in type 2 diabetes (T2D) models but are consistent with other retinopathy models, showing neural damage prior to vascular changes.

show abstract

Diabetic Retinopathy: Retina‐Specific Methods for Maintenance of Diabetic Rodents and Evaluation of Vascular Histopathology and Molecular Abnormalities

Cited by 49 publications

References 57 publications

Photoreceptor Cells Influence Retinal Vascular Degeneration in Mouse Models of Retinal Degeneration and Diabetes

Photoreceptor Cells Influence Retinal Vascular Degeneration in Mouse Models of Retinal Degeneration and Diabetes

Identification of early pericyte loss and vascular amyloidosis in Alzheimer’s disease retina

Characterizing the Retinal Phenotype in the High-Fat Diet and Western Diet Mouse Models of Prediabetes

Contact Info

Product

Resources

About