Photoreceptor degeneration by intravitreal injection of N-methyl-N-nitrosourea (MNU) in rabbits: a pilot study.

ABSTRACT

BACKGROUND: Pilot study on the attempt to induce selective photoreceptor degeneration in the rabbit eye by intravitreal injection of MNU, facing the difficulties of the evaluation of retinal degeneration by different in-vivo and in-vitro methods in such a large eye animal model. METHODS: Eight pigmented Chinchilla Bastard rabbits were injected intravitreally with MNU (1 × 1mg/kg body weight (BW), 1 × 2mg/kg BW, 3 × 3mg/kg BW, 1 × 4mg/kg BW, 1 × 6mg/kg BW, and 1 × DMSO + PBS as control). One, 2, and 3 weeks after injection, the effects on the rabbit retina were examined in vivo using clinical observation (macroscopic images, funduscopy, weighing of the animals), measurement of intraocular pressure (IOP), full-field Electroretinography (ffERG), and spectral-domain Optical Coherence Tomography (sd-OCT). After 3 weeks follow-up, blood samples were taken to evaluate the general health status of the animals, and immunohistochemistry (IH) was performed on sections obtained from six different regions throughout the whole retina to evaluate MNU effects in more detail. RESULTS: It was difficult to observe the effects of MNU on retinal structure by OCT in vivo. Only the temporal quadrant of the retina could be visualized. Therefore, it was indispensible to evaluate the effects of MNU on the retina in vitro by examining six areas of the retina using immunohistochemistry. Furthermore, immunohistochemistry plays a decisive role to evaluate the effects on retinal cells other than photoreceptors while in H&E staining, namely the cell count of the ONL can be observed. The results obtained in vivo and in vitro in this study mainly follow the results of a previous study in mice. The low doses of MNU (1, 2 mg/kg BW) had no effects on retinal function and morphology, while high doses (4, 6 mg/kg BW) led to retinal changes in combination with significant side-effects (e.g., cataractous changes). Injection of 3 mg/kg BW MNU induced selective photoreceptor degeneration. However, the degree of degeneration varied between different parts of the same retina and between retinae of different animals. In two of three animals, a complete loss of ERG potentials was observed. Negative effects on the contralateral eye or on general welfare of the animal were never observed. CONCLUSIONS: In rabbits, the intravitreal injection of 3 mg/kg BW MNU leads to selective but inhomogeneous photoreceptor degeneration.