Proteomic Analysis of the Vitreous following Experimental Retinal Detachment in Rabbits

Abstract

Purpose. The pathogenesis of rhegmatogenous retinal detachment (RRD) remains incompletely understood, with no clinically effective treatment for potentially severe complications such as photoreceptor cell death and proliferative vitreoretinopathy. Here we investigate the protein profile of the vitreous following experimental retinal detachment using a comparative proteomic based approach. Materials and Methods. Retinal detachment was created in the right eyes of six New Zealand red pigmented rabbits. Sham surgery was undertaken in five other rabbits that were used as controls. After seven days the eyes were enucleated and the vitreous was removed. The vitreous samples were evaluated with two-dimensional polyacrylamide gel electrophoresis and the differentially expressed proteins were identified with tandem mass spectrometry. Results. Ten protein spots were found to be at least twofold differentially expressed when comparing the vitreous samples of the sham and retinal detachment surgery groups. Protein spots that were upregulated in the vitreous following retinal detachment were identified as albumin fragments, and those downregulated were found to be peroxiredoxin 2, collagen-Iα1 fragment, and α-1-antiproteinase F. Conclusions. Proteomic investigation of the rabbit vitreous has identified a set of proteins that help further our understanding of the pathogenesis of rhegmatogenous retinal detachment and its complications.

The proteomic profile of a mouse model of proliferative vitreoretinopathy

“…The amount of the Eno1, vimentin, albumin and prohibitin was increased, while the amount of tubulin β‐2C, fructose‐bisphosphate aldolase A and other proteins was reduced in the retina of animals after retinal detachment . In the case of these models the PVR induction was done either by mechanical injury or by retinal detachment surgery and the examined sample was rabbit retina or vitreous .…”

“…The pathophysiological conditions leading to post-translational modifications can only be speculated upon, hypothesizing the effect of MMPs and dispase, respectively, but to best of our knowledge, no experiments aiming to elucidate the phenomena behind the crystallin profile changes have been carried out. The proteomic analysis of rat and rabbit PVR models have revealed the down-regulation of collagen-Ia1, a-1-antiproteinase and peroxiredoxin-2 (Prdx2) in rabbit vitreous, and up-regulation of different forms of a-crystallin A, Tf, b-crystallin B2 (Crybb2), matrix metalloproteinase-3 and other proteins in rabbit retina [39,40]. The amount of the Eno1, vimentin, albumin and prohibitin was increased, while the amount of tubulin b-2C, fructosebisphosphate aldolase A and other proteins was reduced in the retina of animals after retinal detachment [22].…”

“…The level of some intracellular proteins, such as a-enolase (Eno1), tubulin, creatine kinase B-type (Ckb), glutamine synthetase (Glul), G protein, heterogeneous nuclear ribonucleoprotein H (Hnrnph1), heat shock protein b-1 (Hspb1), grifin (Grifin) and fatty-acid binding protein 5, was reduced in the dispase-treated WT samples compared with the physiological saline treated ones (Table 1). In case of spots 11,12,13,14,16,17,18,22 and 24, more than one protein with similar peptide counts reflecting almost equal quantities of each protein was detected. Therefore, the combined effect of these proteins could be monitored in such cases, and no information on the contribution of each individual protein to the spot intensity changes could be observed.…”

Proteins involved in focal adhesion signaling pathways are differentially regulated in experimental branch retinal vein occlusion

“…However, the majority of patients with BRVO do not develop macular edema (Rogers et al, 2010) and protein changes at the retinal level following BRVO remain largely unstudied. Studying large-scale protein changes in BRVO using modern proteomic technologies can bring new insights into pathological mechanisms and elucidate novel biomarkers in BRVO (Cehofski et al, 2014;Honor e and Vorum, 2014;Mandal et al, 2011Mandal et al, , 2013Pollreisz et al, 2013). In the present study label-free liquid chromatography tandem mass spectrometry was used to study large-scale retinal protein changes and associated biological processes following the formation of BRVO in an experimental model.…”

Proteomic analysis of human vitreous associated with idiopathic epiretinal membrane

“…None of these spots changed more than 1.5‐fold, and significantly as determined by Mann–Whitney U test using a significance level at p < 0.05, between the iERM and iMH vitreous groups. High abundance protein spots were excised from the gels for identification with nano‐liquid chromatography‐electrospray ionization‐quadrupole/time‐of‐flight mass spectrometry as previously described (Mandal et al. 2011).…”