Abstract
The mouse model of oxygen-induced retinopathy (OIR) has been widely used in studies related to retinopathy of prematurity, proliferative diabetic retinopathy and in studies evaluating the efficacy of antiangiogenic compounds. In this model, 7-d-old (P7) mouse pups with nursing mothers are subjected to hyperoxia (75% oxygen) for 5 d, which inhibits retinal vessel growth and causes significant vessel loss. On P12, mice are returned to room air and the hypoxic avascular retina triggers both normal vessel regrowth and retinal neovascularization (NV), which is maximal at P17. Neovascularization spontaneously regresses between P17 and P25. Although the OIR model has been the cornerstone of studies investigating proliferative retinopathies, there is currently no harmonized protocol to assess aspects of angiogenesis and treatment outcome. In this protocol we describe standards for mouse size, sample size, retinal preparation, quantification of vascular loss, vascular regrowth, NV and neovascular regression.
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Acknowledgements
This research was generously supported by the V. Kann Rasmussen Foundation, the US National Institutes of Health (EY008670, EY017017, EY14811 (L.E.H.S.); 5 T32 EY07145, 1 F32 EY017789-01 (K.M.C.)), William Randolph Hearst Fund (K.M.C.) and Children's Hospital Boston Mental Retardation and Developmental Disabilities Research Center, P01 HD18655 (L.E.H.S.). Support from the Research to Prevent Blindness Lew Wasserman Merit Award (L.E.H.S). Juvenile Diabetes Research Foundation International (3-2006-278, 10-2008-603 (J.C.). The sponsors had no role in the design or conduct of the study, in the collection, analysis and interpretation of data and in the preparation, review or approval of the manuscript.
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K.M.C. wrote the manuscript with N.M.K., R.J.D. and L.E.H.S. with input from C.M.A., J.C., K.G., P.S., A.S. and K.L.W. All authors have worked on and modernized this protocol.
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Supplementary information
Supplementary Video 1
Retinal Dissection Movie (MOV 156087 kb)
Supplementary Video 2
Movie of retinal flatmount (MOV 87767 kb)
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Connor, K., Krah, N., Dennison, R. et al. Quantification of oxygen-induced retinopathy in the mouse: a model of vessel loss, vessel regrowth and pathological angiogenesis. Nat Protoc 4, 1565–1573 (2009). https://doi.org/10.1038/nprot.2009.187
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DOI: https://doi.org/10.1038/nprot.2009.187
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