Diabetes triggers slight immunoreactive changes in tight junction proteins, without changes in retinal vascular permeability. (IMAGE)
Caption
Diabetes triggers slight immunoreactive changes in tight junction proteins, without changes in retinal vascular permeability. Diabetic Wistar Han rats (induced by a 12-week HFD with STZ injection at week 4; 35 mg/kg, IP; T2D group) presented a slight decrease in claudin-5 immunoreactivity after 8 weeks on HFD, as well as in occludin and claudin-5 immunostaining at week 12, as assessed in retinal wholemounts. However, no differences were found in its protein levels in retinal extracts, assessed by Western blot, compared to age-matched controls (Control group). No vascular leakage was detected in diabetic animals, as assessed by the Evans blue assay. Representative images of retinal wholemounts immunostained for (a) claudin-5, (b) occludin, and (c) ZO-1. (d) Representative images showing Evans blue fluorescence in the retina. (e) Claudin-5, (f) occludin, and (g) ZO-1 protein levels assessed by Western blot, normalised to the loading control (calnexin), and expressed as percentage of the respective control. Representative images of protein immunoreactive bands are presented above the graphs, with the respective loading control (calnexin). Data are presented as mean ± SEM. Statistical analysis was performed using the Student’s t-test, or the Mann–Whitney test, if data were not normally distributed. HFD, high-fat diet; STZ, streptozotocin; IP, intraperitoneal; T2D, type 2 diabetes; ZO-1, zonula occludens-1.
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Eye and Vision
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