Estudo eletrofisiológico de modelo animal para retinopatia da prematuridade

Panassiol, R. T. (2017). Electrophysiological Study of an Animal Model for Retinopathy of Prematurity (Master ́s Thesis). Instituto de Psicologia, Universidade de São Paulo, São Paulo. The retinopathy of prematurity is an ocular developmental disorder associated with abnormal retinal vascular growth and occurs only in neonates younger than 32 weeks of gestation and undergoing long periods in incubators, typically rich in oxygen. The disease has two phases: (1) the inhibition of the normal development of vessels in the retina, due to the hyperoxia induced by the incubator environment; (2) retinal neovascularization due to the increase in growth factors triggered by the low availability of oxygen outside the incubator. The objective of this study is to evaluate the visual function in an animal model of retinopathy of prematurity, comparing it to healthy animals. Full-field electroretinogram (ERGs) were performed on control mice (Mus musculus) and that were exposed to a hyperbaric chamber (75% oxygen) during post-natal retinal development to understand the visual losses that occur in retinopathy of prematurity. We used 122 animals, divided into two groups: in the control group, ERGs were performed at P17 (n = 32), P30 (n = 26) and ≥P60 (n = 44) with stimulation at different wavelengths (LEDs with peak of emission at 365 nm, 459 nm, 525 nm and 635 nm) for evaluation of the functional activity of the retina throughout the development, without any additional experimental manipulation; in the experimental group, animals were submitted to hyperoxogenation of P7 to P12 for induction of angiogenesis and evaluated with ERGs with stimulation at 459 nm in P17 (n = 7), P30 (n = 6) and ≥P60 (n = 7) to follow up retinal development. All animals were dark adapted for at least two hours prior to the experiments. In sessions with 40 to 60 minutes, the animals were subjected to flashes of light at increasing intensities. Amplitudes and latencies of a-waves, b-waves and oscillatory potentials were measured, and intensity-response relationships adjusted with different mathematical models for comparison between individuals and between groups. The parameters obtained with the adjustments were compared through ANOVA and Student's T tests with the appropriate Bonferroni correction. In the light levels tested, the murine ERG b-wave is dominated by rod activity and the a-wave is dominated by the cones. The records in animals at P17 and P30 of the control group were similar to those performed at ≥P60 regarding the latency and amplitude of response, as well as the sensitivity and degree of cooperation between the different ERG elements. However, the maximum response amplitudes were slightly higher at P30 at all wavelengths and there was slight reduction of absolute sensitivity throughout development. For the experimental group, the animals at P17 suffered the greatest losses, with decreases in the amplitudes of all the components of the ERG, without prejudice to the latencies or sensitivity. There was also a successive recovery of responses throughout animal development. These findings indicate that the ROP model in mice reproduces aspects essential to the severe human pathology.