Investigation of the role of Glil+ cells in retinal angiogenesis and fibrosis.
type: CreativeWork
description: Background: In ocular diseases such as Age-related Macular Degeneration (AMD) and diabetic retinopathy, dysregulated inflammation in the retina can lead to the formation of a fibrovascular membrane, which can cause severe structural damage to the retina thus leading to severe visual defects and blindness. The process of pathological neovascularisation into the formation of a fibrovascular membrane is not completely understood. Recent research has implicated the Hedgehog pathway transcription factor, Gli1, in the development of fibrosis in numerous organs. This study aims to characterise Gli1 expression in ocular tissues and to elucidate the role of Gli1 in the development of fibrovascular membranes in the retina utilising transgenic mice, and specific small molecule inhibitor of Gli1, GANT61. Methods: Gli1CreERT2 mice were crossed with Ai9(RCL-tdT) (Tomato) mice to produce Gli1-Cre x Tomato crossbreeds. 6–7-week-old Heterozygous Gli1-Cre x Tomato mice received 100 mg/kg Tamoxifen daily (I.P.) for 5 days. Mice were monitored for 80 days to determine the duration Gli1-Tomato expression. Eyes, and other organs, were collected for immunofluorescent staining. For the one laser choroidal neovascularisation (CNV) model, Gli1-Cre x Tomato mice received 3-4 laser burns per eye to induce choroidal neovascularisation and were monitored over 21 days to assess the expression of Gli1 in the injured retina using fundus imaging and immunofluorescent staining. For the two-stage CNV model, Gli1-Cre x Tomato mice received 3-4 laser burns per eye to induce choroidal neovascularisation followed by a second laser directed towards CNV lesions 7 days later. Mice were then monitored over a period of 30 days. Two-stage laser mice received either GANT61 or diphtheria toxin (for Gli1-cre x Rosa mice) to determine the effects of Gli1 inhibition/ablation on fibrovascular lesion development. Cell survival assays of retinal pigment epithelial and microvascular endothelial cells in the presence of …
abstract: Background: In ocular diseases such as Age-related Macular Degeneration (AMD) and diabetic retinopathy, dysregulated inflammation in the retina can lead to the formation of a fibrovascular membrane, which can cause severe structural damage to the retina thus leading to severe visual defects and blindness. The process of pathological neovascularisation into the formation of a fibrovascular membrane is not completely understood. Recent research has implicated the Hedgehog pathway transcription factor, Gli1, in the development of fibrosis in numerous organs. This study aims to characterise Gli1 expression in ocular tissues and to elucidate the role of Gli1 in the development of fibrovascular membranes in the retina utilising transgenic mice, and specific small molecule inhibitor of Gli1, GANT61. Methods: Gli1CreERT2 mice were crossed with Ai9(RCL-tdT) (Tomato) mice to produce Gli1-Cre x Tomato crossbreeds. 6–7-week-old Heterozygous Gli1-Cre x Tomato mice received 100 mg/kg Tamoxifen daily (I.P.) for 5 days. Mice were monitored for 80 days to determine the duration Gli1-Tomato expression. Eyes, and other organs, were collected for immunofluorescent staining. For the one laser choroidal neovascularisation (CNV) model, Gli1-Cre x Tomato mice received 3-4 laser burns per eye to induce choroidal neovascularisation and were monitored over 21 days to assess the expression of Gli1 in the injured retina using fundus imaging and immunofluorescent staining. For the two-stage CNV model, Gli1-Cre x Tomato mice received 3-4 laser burns per eye to induce choroidal neovascularisation followed by a second laser directed towards CNV lesions 7 days later. Mice were then monitored over a period of 30 days. Two-stage laser mice received either GANT61 or diphtheria toxin (for Gli1-cre x Rosa mice) to determine the effects of Gli1 inhibition/ablation on fibrovascular lesion development. Cell survival assays of retinal pigment epithelial and microvascular endothelial cells in the presence of …
author: Marry, Stephen
language: eng
keywords: Hedgehog Signalling, Gli1, Fibrosis, CNV, retina, retinal microvascular, neovascularisation, laser-induced neovascularisation, two-staged laser model
date published: 2022-7
date modified: 2026-07-24T03:56:18Z
provider: Queen's University Belfast
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: Queen's University Belfast