Desiccating Stress Dry Eye Model in Mice

All studies performed were approved by The Institutional Animal Care and Use Committees at Baylor College of Medicine and comply with the ARVO Statement for the Use of Animals in Vision Research. Female C57BL/6 mice aged 6–8 weeks old were purchased from Jackson Laboratories (Bar Harbor, ME). DS was induced by subcutaneous injection of scopolamine hydrobromide (0.5 mg/0.2 ml; Sigma-Aldrich, St. Louis), QID (08:00, 12:00, 14:00, and 17:00 h), for 3, 5 or 10 consecutive days in 6–8 week old female C57BL/6 mice and euthanized at the end of each variable (DS3, DS5 and DS10), as previously published (de Paiva et al., 2009 (link); de Paiva et al., 2006a (link); de Paiva et al., 2006b (link)). Mice were placed in a cage with a perforated plastic screen on one side to allow airflow from a fan placed six inches in front of it for 16 h/day. Room humidity was maintained at 30–35%. Control mice were maintained in a non-stressed (NS) environment containing 50–75% relative humidity without exposure to forced air. Since dry eye is more prevalent in women and male mice do not respond well to desiccation, only female mice were used (Gao et al., 2015 (link); Schaumberg et al., 2003 (link), 2009 (link)).