Abstract
Purpose: :
We have previously shown that the isoprostane, 8-isoprostaglandin F2 (8-isoPGF2) can regulate neurotransmitter release from mammalian neural retina, in vitro (Opere et al.,Neurochem. Res.2005). However, the effect this isoprostane on retinal protein expression has not been determined. Experiments in this study were designed to investigate the effects of intravitreally injected 8-isoPGF2 on protein expression in bovine retina, ex vivo.
Methods: :
Freshly isolated bovine eyeballs were injected intravitreally with 8-isoPGF2 and neural retina were isolated and prepared for determination of protein expression using the 2DE-DIGE approach. Protein spots exhibiting statistically significant changes (P<0.05) were selected and identified using LC-MS/MS and analyzed by bioinformatics.
Results: :
8-isoPGF2 (10uM) up-regulated the expression of two proteins while nine were down-regulated. Specifically, the expression of triosephosphate isomerase which is involved in energy metabolism was increased. On the contrary, expression of proteins involved in cellular defense: peroxiredoxin 2, a scavenger for peroxides; aldehyde dehydrogenase 9 family, member A1, an enzyme that metabolizes alcohol; heat shock cognate 71, a protein involved in cellular defense against hypoxia were down-regulated. Furthermore, the expression of ATPase, H+ transporter; GDP dissociation inhibitor 2, a signal transduction protein; calretinin, a calcium binding protein that is used as a marker for neuronal subpopulations and recoverin, a calcium-binding protein in photoreceptor cells whose antibody has been linked to some retinopathies were also down-regulated.
Keywords: eicosanoids • proteomics • retina