Mouse and Rat Oxygen-Induced Retinopathy Models to Study Vascular Features Seen in Retinopathy of Prematurity.

Retinopathy of prematurity (ROP), a retinovascular disease, is a leading cause of childhood blindness worldwide. Given the constraints of studying molecular mechanisms in preterm infants, reproducible animal models are important to understand ROP pathophysiology. Mouse and rat oxygen-induced retinopathy (OIR) models are the most commonly used and recapitulate key vascular features seen in ROP. However, these models are susceptible to inherent variability that limits reproducibility, including inter-litter variability, consistency of oxygen delivery across experiments, retinal dissection technique, and immunohistochemistry. Here, we describe a comprehensive protocol for performing the most common mouse and rat OIR models, and procedures such as eye enucleation, retinal dissection and flat mounting, isolectin GS-IB4 staining, whole retina stitched fluorescence imaging from Z-stacks, and quantification of vascular features. This protocol provides important materials and procedural details to increase the reproducibility of the mouse and rat OIR models. Key features • Rat and mouse oxygen-induced retinopathy (OIR) models. • Eye enucleation of experimental rat and mouse pups. • Retinal flat mounting and immunostaining for rat and mouse eyes. • Image analysis of retinal flat mounts from rat and mouse eyes.