Acepromazine

For surgical procedures and euthanasia, animals were anesthetized with an intraperitoneal injection of 50 mg/kg ketamine (Fort Dodge Animal Health, Fort Dodge, IA), 10 mg/kg xylazine (VedCo Inc., Saint Joseph, MO), and 2 mg/kg acepromazine (Phoenix Pharmaceuticals, Burlingame, CA), and received topical ocular anesthesia of 0.5% proparacaine hydrochloride (Akorn Inc. Buffalo Grove, IL, USA). For IOP measurements independent of additional procedures, animals were anesthetized using a Rodent Circuit Controller (VetEquip, Inc., Pleasanton, CA, USA) delivering 2.5% isoflurane in oxygen at 500 cc/min.

In the glaucoma experiment, 200 mice were anesthetized with a mixture of ketamine (Fort Dodge Animal Health, Fort Dodge, IA), xylazine (VedCo Inc., Saint Joseph, MO), and acepromazine (Phoenix Pharmaceuticals, Burlingame, CA) at 50, 10 and 2 mg/kg, respectively. The dosage and time of anesthesia was controlled and has been standardized as previously published [11 (link)]. Then, one anterior chamber was injected with Polybead Microspheres^® (Polysciences, Inc., Warrington, PA, USA), using the 4+1 protocol [11 (link)], consisting of 2 μl of 6 μm diameter beads, then 2 μl of 1 μm diameter beads, followed by 1 μl of viscoelastic compound (10 mg/ml sodium hyaluronate, Healon; Advanced Medical Optics Inc., Santa Ana, CA). The injections were made through a glass cannula with 50 μm tip diameter, connected to a Hamilton syringe (Hamilton, Inc., Reno, NV). IOP was measured immediately after injection, and at 3 days, 1 week, 2 weeks and 6 weeks, using the Tonolab tonometer. Investigators were masked as to treatment group during IOP measurements. The experimental and control animals were treated in masked fashion on the same days, interchangeably, so that any difference in the state or effect of anesthesia would be random and would not lead to any systematic bias. At all steps of the experiment, the solutions and tissues were coded to mask the participants.

For surgical procedures and euthanasia, animals were anesthetized with an intraperitoneal injection of 50 mg/kg ketamine (Fort Dodge Animal Health, Fort Dodge, IA, USA), 10 mg/kg xylazine (VedCo Inc., Saint Joseph, MO, USA), and 2 mg/kg acepromazine (Phoenix Pharmaceuticals, Burlingame, CA, USA), and received topical ocular anesthesia of 0.5% proparacaine hydrochloride (Akorn Inc., Buffalo Grove, IL, USA). For IOP measurements, independent of additional procedures, animals were anesthetized using a Rodent Circuit Controller (VetEquip, Inc., Pleasanton, CA, USA) delivering 2.5% isoflurane in oxygen at 500 cc/min.