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Updated: Mar 18, 2026
Transpupillary Two-Photon In Vivo Imaging of the Mouse Retina
Published on: February 13, 2021
Differential expression of microRNAs in retinal vasculopathy caused by selective Müller cell disruption
Sook Hyun Chung1, Mark Gillies1, Michelle Yam1
1Macula Research Group, Clinical Ophthalmology and Eye Health, Save Sight Institute, the University of Sydney, Sydney, Australia.
Abstract:
Vascular changes and photoreceptor degeneration are features of age-related macular degeneration, diabetic retinopathy and macular telangiectasis. We have profiled the differential expression of microRNAs and analysed their target genes in transgenic mice in which induced Müller cell disruption results in photoreceptor degeneration, vascular leak and deep retinal neovascularisation. We identified 9 miRNAs which were differentially expressed during the development of retinal neovascularization and chose miR-200b and its target genes for further study. Using qRT-PCR and western blot analysis, we found that downregulation of miR-200b was negatively correlated with its target genes, including zinc finger E-box binding homeobox (ZEB) 1 and 2 and vascular endothelial growth factor receptor 1. Double immunofluorescence labelling revealed that the newly formed vessels in the outer retina were positive for ZEB2. Furthermore, intravitreal injections of a miR-200b-mimic and anti-miR-200b confirmed the negative correlation of miR-200b and its target gene expression. We also found that the miR-200b-mimic inhibited vascular leak in the established mild vascular lesions, whereas anti-miR-200b promoted it. Taken together, these data suggest that miR-200b may play a role in the development of intraretinal neovascularisation.
Insights
MicroRNA-200b (miR-200b) downregulation is linked to retinal neovascularization and vascular leak in mice. Restoring miR-200b levels may inhibit new blood vessel formation in the retina.
Area of Science:
- Ophthalmology
- Molecular Biology
- Genetics
Background:
- Age-related macular degeneration, diabetic retinopathy, and macular telangiectasis involve vascular changes and photoreceptor degeneration.
- Müller cell disruption in mice models photoreceptor degeneration, vascular leak, and retinal neovascularization.
Purpose of the Study:
- To investigate the role of microRNAs (miRNAs) and their target genes in retinal neovascularization.
- To identify specific miRNAs involved in the development of retinal neovascularization and vascular leak.
Main Methods:
- Differential expression profiling of miRNAs in a mouse model of retinal neovascularization.
- Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analysis to assess miRNA and target gene expression.
- Intravitreal injections of miR-200b-mimic and anti-miR-200b to evaluate functional effects.
Main Results:
- Nine differentially expressed miRNAs were identified; miR-200b and its targets (ZEB1, ZEB2, VEGFR1) were selected for further study.
- Downregulation of miR-200b correlated negatively with its target genes ZEB1, ZEB2, and VEGFR1.
- ZEB2 was found in newly formed vessels in the outer retina.
- miR-200b-mimic inhibited vascular leak, while anti-miR-200b promoted it.
Conclusions:
- miR-200b plays a significant role in regulating intraretinal neovascularization.
- The miR-200b/ZEB pathway is implicated in retinal vascular pathologies.
