Subretinal injections in rodent eyes: Effects onelectrophysiology and histology of rat retina

[摘要] Purpose: To describe a reliable and fast method for subretinalinjection in rodents and to assess the effect of the procedure onretinal function and histology.Methods: Corneas of rodents were punctured with a 28 gaugehypodermic insulin needle avoiding the lens. The injection procedure canbe observed with the aid of a dissecting microscope and methylcellulosesolution on the eye. A 33 gauge blunt needle was inserted into the eyethrough the corneal puncture and guided toward the subretinal matrix.Addition of fluorescein to the injection mixture facilitated immediateevaluation of the injection. Rat eyes were either non-injected(controls), received only a corneal puncture or were injected withfluorescent microspheres or PBS-fluorescein mixture. Retinal functionand integrity were assessed through electroretinographic (ERG) analysisand postmortem histology.Results: The anterior injection procedure provided a fast and simplemethod for subretinal injections. In rats a successful subretinaldelivery was achieved in more than 90%, with less than 5% of theinjected eyes developing cataracts. No significant differences in b-waveERG amplitudes in rodent eyes over a five-week period were observedbetween non-injected control eyes and subretinally injected eyes (1 to10 ml of PBS-fluorescein or 2 ml fluorescent microspheres).Histological analysis revealed that re-attachment of the rat retinaoccurred in approximately 1 day post-injection and the phagocytoticability of RPE cells remained intact.Conclusions: This method was easily learned and required a minimumof equipment and animal preparation. With experience, 10 to 30 eyescould be injected per h. Furthermore, the injection procedure did notcompromise the lens, retina or retinal pigment epithelium (RPE).

[学科分类] 生物化学/生物物理