Purchase this article with an account.
Francois Paquet-Durand, Stavros Vagionitis, Ksenija Martinovic, Joaquin Valdes, Dragana Trifunovic, Maria Miranda, Oliver Schmachtenberg; Simulating diabetic retinopathy in organotypic retinal explant cultures: Comparison of diabetic conditions on early vs. late post-natal retina. Invest. Ophthalmol. Vis. Sci. 2017;58(8):5192.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Diabetic retinopathy (DR) is one of the leading causes of vision impairment worldwide. Unfortunately, DR research is hindered by a lack of disease models that faithfully reproduce the retinal phenotype of diabetes, in particular for type 2 diabetes. Previously, we have shown that a number of aspects of DR can be faithfully reproduced in early (P5) post-natal organotypic retinal explant cultures (Valdés et al., ALTEX 33:459-464, 2016). However, since the retina is not fully developed at this stage, it remains unclear whether the deleterious effects of diabetes-like conditions on the early postnatal retina are representative of mature retinal pathophysiology.
Mouse retinas were explanted at P14 (around eye opening) and treated from P16 to different with different experimental conditions (i.e. no-insulin, high-glucose, no-insulin + high-glucose, 2-deoxyglucose; 2-DG) to simulate type 1 and 2 diabetic conditions. The treatment effects were assessed on histological preparations using photoreceptor row counts, TUNEL assay for cell death detection, and immunostaining for cone arrestin.
In the no-insulin, high-glucose, and no-insulin + high-glucose treatment paradigms the photoreceptor row counts, numbers of TUNEL, and cone arrestin-positive cells did not change significantly when compared to control. However, the 2-DG treatment significantly increased the numbers of dying TUNEL-positive cells in the outer nuclear layer (ONL; control: 1.22% ± 0.3 STD; 2-DG: 13.76% ± 0.8, p<0.001) and reduced the number of arrestin-positive cones per 100 µM of retinal circumference (control: 4.7 ± 0.7; 2-DG: 0.4 ± 0.3, p<0.001). Yet, the overall photoreceptor row count was not significantly altered (control: 8.0 ± 0.6; 2-DG: 8.1 ± 0.4).
Compared to the early post-natal retina, older retinas responded less strongly to simulated diabetic conditions, yet, cone photoreceptors were highly vulnerable to type I diabetes-like conditions caused by 2-DG treatment. Taken together, we have further validated and extended an in vitro model of DR which may prove useful for studies into the etiopathology of DR and for DR-related drug screening.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
This PDF is available to Subscribers Only