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Generation of transgenic mice with mild and severe retinal neovascularisation
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  1. C-M Lai1,
  2. S A Dunlop2,3,
  3. L A May1,
  4. M Gorbatov4,
  5. M Brankov1,4,
  6. W-Y Shen1,4,
  7. N Binz1,4,
  8. Y KY Lai1,4,
  9. C E Graham1,4,
  10. C J Barry1,4,
  11. I J Constable1,
  12. L D Beazley2,3,
  13. E P Rakoczy1
  1. 1Centre for Ophthalmology and Visual Science, The University of Western Australia, Nedlands, Australia
  2. 2School of Animal Biology, The University of Western Australia, Nedlands, Australia
  3. 3Western Australian Institute for Medical Research, The University of Western Australia, Nedlands, Australia
  4. 4Lions Eye Institute, Molecular Ophthalmology Department, Nedlands, Australia
  1. Correspondence to: Associate Professor Chooi-May Lai Lions Eye Institute, 2 Verdun Street, Nedlands, Western Australia, 6009, Australia; mlaicyllene.uwa.edu.au

Abstract

Aim: To generate a mouse model for slow progressive retinal neovascularisation through vascular endothelial growth factor (VEGF) upregulation.

Methods: Transgenic mice were generated via microinjection of a DNA construct containing the human VEGF165 (hVEGF) gene driven by a truncated mouse rhodopsin promoter. Mouse eyes were characterised clinically and histologically and ocular hVEGF levels assayed by ELISA.

Results: One transgenic line expressing low hVEGF levels showed mild clinical changes such as focal fluorescein leakage, microaneurysms, venous tortuosity, capillary non-perfusion and minor neovascularisation, which remained stable up to 3 months postnatal. Histologically, there were some disturbance and thinning of inner and outer nuclear layers, with occasional focal areas of neovascularisation. By contrast, three other lines expressing high hVEGF levels presented with concomitantly severe phenotypes. In addition to the above, clinical features included extensive neovascularisation, haemorrhage, and retinal detachment; histologically, focal to extensive areas of neovascularisation associated with retinal folds, cell loss in the inner and outer nuclear layers, and partial retinal detachment were common.

Conclusions: The authors generated four hVEGF overexpressing transgenic mouse lines with phenotypes ranging from mild to severe neovascularisation. These models are a valuable research tool to study excess VEGF related molecular and cellular changes and provide additional opportunities to test anti-angiogenic therapies.

  • CFP, colour fundus photography
  • CNV, choroidal neovascularisation
  • DR, diabetic retinopathy
  • ELISA, enzyme linked immunosorbent assay
  • FFA, fluorescein fundus angiography
  • FITC, fluorescein isothiocyanate
  • hVEGF, human vascular endothelial growth factor
  • INL, inner nuclear layer
  • NFL, nerve fibre layer
  • NPDR, non-proliferative diabetic retinopathy
  • ONL, outer nuclear layer
  • PBS, phosphate buffered saline
  • PCR, polymerase chain reaction
  • PDR, proliferative diabetic retinopathy
  • RPE, retinal pigment epithelium
  • ROP, retinopathy of prematurity
  • VEGF, vascular endothelial growth factor
  • transgenic mouse model
  • retinal neovascularisation
  • vascular endothelial growth factor
  • CFP, colour fundus photography
  • CNV, choroidal neovascularisation
  • DR, diabetic retinopathy
  • ELISA, enzyme linked immunosorbent assay
  • FFA, fluorescein fundus angiography
  • FITC, fluorescein isothiocyanate
  • hVEGF, human vascular endothelial growth factor
  • INL, inner nuclear layer
  • NFL, nerve fibre layer
  • NPDR, non-proliferative diabetic retinopathy
  • ONL, outer nuclear layer
  • PBS, phosphate buffered saline
  • PCR, polymerase chain reaction
  • PDR, proliferative diabetic retinopathy
  • RPE, retinal pigment epithelium
  • ROP, retinopathy of prematurity
  • VEGF, vascular endothelial growth factor
  • transgenic mouse model
  • retinal neovascularisation
  • vascular endothelial growth factor

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Footnotes

  • Sponsor details. This study was supported by the Juvenile Diabetes Research Foundation International (USA), National Health and Medical Research Council (Australia) and Westpac Foundation (Australia).

  • Ethical approval. Ethical approval (AEC 03/300/016 and 03/100/107) for breeding of and experimentation on animals has been granted by the Animal Ethics Committee at the University of Western Australia, Australia.

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