Laser-Induced Rat CNV Lesion Protocol

The CNV lesions were induced in rat eyes by laser photocoagulation as previously described [26 (link)]. Briefly, Brown Norway rats were anesthetized with an intramuscular injection of a mixture of 2% xylocaine (0.15 mL/kg body weight, Astra, Astra Sodertalje, Sweden) and ketamine (50 mg/kg body weight, Parke-Davis, Morris Plains, NJ, USA). Pupils were dilated with 1% tropicamide (Alcon Laboratories, Fort Worth, TX, USA). A piece of cover glass served as the contact lens to improve the visibility of the fundus. Argon laser (Novus Omni; Coherent, Palo Alto, CA, USA) irradiation was delivered through a slit lamp (Carl Zeiss, Oberkochen, Germany). Laser parameters: spot size of 50 µm, power of 400 mW, and exposure duration of 0.05 s. Disruption of Bruch’s membrane was detected by the emergence of a bubble at the center of photocoagulation in the laser spotted zone. Six lesions were generated in each eye at the 1, 3, 5, 7, 9, and 11 o’clock positions located at equidistance from the optic disk and between the major retinal vessels.