Diabetic Mouse Cornea Experiments

In this study, 7-week-old male C57BL/6 mice were purchased from CLEA Japan Inc. (Tokyo, Japan). STZ (Sigma-Aldrich, St. Louis, Missouri) (200 mg/kg) was intraperitoneally administered once in 8-week-old male C57BL/6 mice to induce diabetes mellitus in mice⁵⁵ . Diabetic mice with blood glucose levels > 300 mg/dL were used in the experiments. The AGE inhibitor pyridoxamine dihydrochloride (Tokyo Chemical Industry, Tokyo, Japan) in 1 g/L of water was used to suppress AGE deposition in the cornea of diabetic mice. Male C57BL/6 mice began consuming this water 1 week before the intraperitoneal administration of STZ^{37 (link)}. We raised these mice until 24 weeks of age and used them for experiments. Diabetic mice (24-week-old), diabetic mice that consumed pyridoxamine (24-week-old), age-matched wild-type mice (24-week-old), and young wild-type mice (8-week-old) were deeply anesthetized with a cocktail of medetomidine hydrochloride (1.0 mg), midazolam (5.0 mg/mL), and butorphanol (5.0 mg/mL). After anesthesia induction, the mice were euthanized by de-bleeding from the axillary artery, and only the cornea was removed and used in the experiments.