Intracameral Adeno-Associated Virus Delivery

Three-month-old C57BL/6J wild-type or FP receptor knockout mice previously generated by our laboratory [39 (link)] were anesthetized with an intraperitoneal injection of ketamine/xylazine/acepromazine (80/6/1 mg/kg body weight) for intracameral injections. Mice were subsequently placed on a dissecting microscope to visualize the anterior chamber. A 32 gauge beveled needle (Hamilton Company, Reno, NV) containing the AAV2 construct (1 μL; 3 x 10¹² VG/mL) or 1 μL phosphate buffered saline (PBS) was placed parallel to the cornea and inserted anterior to the limbus into the anterior chamber. Care was taken to avoid iris trauma as well as avoidance of the corneal endothelium and anterior lens capsule. The injected volume was administered into the anterior chamber over 2 seconds. The needle was then slowly removed to minimize reflux.