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Retinal vein occlusion induced by laser treatment in mice

ABSTRACT: Purpose: Several animal models are available for investigating retinal vein occlusion (RVO). Laser-induced RVO has been studied in pigs, rabbits, and rats. Mice have been rarely used despite the huge number of available mutants, ease of handling and cost effectiveness. The aim of this study was to find out if the RVO mouse model is useful for investigating effects of hypoxia. Methods: C57Bl/6J mice were injected with eosin Y for photo-sensitization. Subsequently, large retinal veins were laser-treated in one eye to induce vascular occlusion. Contralateral control eyes received non-occlusive laser treatment of the retina sparing large vessels. The animals were followed for up to eight days and assessed by fundoscopy, angiography, hypoxyprobe staining, histopathology and gene expression analysis by qPCR and RNA sequencing. Another group of mice was left untreated and studied at a single time point to determine baseline characteristics. Results: In the RVO group, half of the retinal veins stayed occluded for three days, and one third did so for the whole test period of 8 days. Hypoxia was observed in all RVO eyes for up to 5 days with a maximum around days 2-3. Large retinal edematous areas were present in all laser-treated eyes irrespective of vascular occlusion status. Accordingly, expression of genes associated with inflammation or cell damage was highly up-regulated in both groups as compared to untreated eyes while expression of genes associated with angiogenesis or hypoxia did not change much. Conclusion: In the laser-induced RVO mouse model, general tissue damage and inflammation responses predominate and obscure specific hypoxia- or angiogenesis-related changes. A non-occlusive control laser treatment is essential to allow for proper data interpretation.

ORGANISM(S): Mus musculus

PROVIDER: GSE101398 | GEO | 2018/03/14

REPOSITORIES: GEO

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Project description:Human CD34+ stem cells are mobilized from bone marrow to sites of tissue ischemia and play an important role in tissue revascularization. This study used a murine model to test the hypothesis that intravitreal injection of human CD34+ stem cells harvested from bone marrow (BMSCs) can have protective effects in eyes with diabetic retinopathy. Streptozotocin-induced diabetic mice (C57BL/6J) were used as a model for diabetic retinopathy. Subcutaneous implantation of Alzet pump, loaded with Tacrolimus and Rapamycin, 5 days prior to intravitreal injection provided continuous systemic immunosuppression for the study duration to avoid rejection of human cells. Human CD34+ BMSCs were harvested from the mononuclear cell fraction of bone marrow from a healthy donor using magnetic beads. The CD34+ cells were labeled with enhanced green fluorescent protein (EGFP) using a lentiviral vector. The right eye of each mouse received an intravitreal injection of 50,000 EGFP-labeled CD34+ BMSCs or phosphate buffered saline (PBS). Simultaneous multimodal in vivo retinal imaging system consisting of fluorescent scanning laser ophthalmoscopy (enabling fluorescein angiography), optical coherence tomography (OCT) and OCT angiography was used to confirm the development of diabetic retinopathy and study the in vivo migration of the EGFP-labeled CD34+ BMSCs in the vitreous and retina following intravitreal injection. After imaging, the mice were euthanized, and the eyes were removed for immunohistochemistry. In addition, microarray analysis of the retina and retinal flat mount analysis of retinal vasculature were performed. The development of retinal microvascular changes consistent with diabetic retinopathy was visualized using fluorescein angiography and OCT angiography between 5 and 6 months after induction of diabetes in all diabetic mice. These retinal microvascular changes include areas of capillary nonperfusion and late leakage of fluorescein dye. Multimodal in vivo imaging and immunohistochemistry identified EGFP-labeled cells in the superficial retina and along retinal vasculature at 1 and 4 weeks following intravitreal cell injection. Microarray analysis showed changes in expression of 162 murine retinal genes following intravitreal CD34+ BMSC injection when compared to PBS-injected control. The major molecular pathways affected by intravitreal CD34+ BMSC injection in the murine retina included pathways implicated in the pathogenesis of diabetic retinopathy including Toll-like receptor, MAP kinase, oxidative stress, cellular development, assembly and organization pathways. At 4 weeks following intravitreal injection, retinal flat mount analysis showed preservation of the retinal vasculature in eyes injected with CD34+ BMSCs when compared to PBS-injected control. The study findings support the hypothesis that intravitreal injection of human CD34+ BMSCs results in retinal homing and integration of these human cells with preservation of the retinal vasculature in murine eyes with diabetic retinopathy.

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Retinal vein occlusion induced by laser treatment in mice

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