Brown norway rat

Procedures concerning animals were performed in accordance with the EU Directive 2010/63/EU for animal experiments and with permission of the Dutch Central Authority for Scientific Procedures on Animals (CCD), permit number 1160020172924, approved 18, January, 2018. The animals were maintained on a 12 h day–night cycle and were supplied ad libitum with food and water. Brown Norway rats from Janvier Labs with a spontaneous mutation in the Crb1 gene were used in this study [10 (link)]; the Crb1 mutant rat breeding was set up within the LUMC animal facility. Age-matched control Brown Norway rats lacking the mutation in the Crb1 gene from Charles River Laboratories were used as controls. Animals were killed by carbon dioxide inhalation.

Animal experiments were performed in accordance with the Association for Research in Vision and Ophthalmology (ARVO) Statement concerning the use of animals in ophthalmic and vision research. The experimental procedure was approved and monitored by the Animal Care and Use Committee of the Nara Research & Development Center, Santen Pharmaceutical Co., Ltd. Six to eight-week-old female Brown Norway rats were purchased from Charles River Japan (Yokohama, Japan). Rats were housed under a 12-hour light/12-hour dark cycle and provided with food and water ad libitum.

A total of fifty-two adult Lewis rats (300 g, Charles Rivers Laboratories) and eight Brown-Norway rats (350 g, Charles River Laboratories) were used for this study. This study was designed and carried out in accordance with the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. This protocol was approved by the Johns Hopkins University Animal Care and Use Committee (RA11M130). All animals were housed in a central animal care facility with a 12-hour light/12-hour dark cycle, and provided with adequate food/water ad libitum.

Two-month-old Brown Norway rats weighing approximately 200 g (g) were obtained from Charles River Laboratories (Wilmington, MA). All the animals were treated according to the ARVO Statement for the Use of Animals in Ophthalmology and Vision Research. After 12 h (hr) fasting, the animals received a single 50 mg/kg intraperitoneal (IP) injection of STZ (Sigma-Aldrich, St. Louis, MO) in 10 mM of sodium citrate buffer, pH 4.5. Control (non-diabetic) animals were fasted and injected with the citrate buffer alone. Animals with blood glucose levels greater than 300 mg/dl were considered diabetic and randomly divided into groups. Diabetic rats at 2 weeks after onset of diabetes received an intravitreal injection of PEDF34-NP (20 µg/eye) in the treatment group and the same numbers of empty PGLA-NP (Control-NP) in the control group.

Three-month-old male and female Brown Norway rats (Charles River Laboratories) were used. All rats were kept in 12 hours of white fluorescent light and 12 hours of darkness for the first 3 months of life. Fluorescent lamps provided white light (≤ 100 lx). Then, the acute exposure (AE) group (9 rats) was first kept in total darkness for 24 hours, and then continuously exposed to blue light for 48 hours. The long-term exposure (LTE) group (9 rats) was exposed to 12 hours of blue light and 12 hours of darkness for 10 days. The control group (6 rats) was kept in 12 hours of white light (1000 lx) and 12 hours of darkness for 10 days. To dilate the rats’ pupils in all 3 groups, 1% atropine drops were applied twice a day.
All rats were kept in transparent, plexiglass cages (Supplementary Fig. S1). Blue light was provided by clusters of light emitting diodes (LEDs; 463 ± 10 nm; 400 blue diodes per cage; 50 diodes provide 16,000 lumen/5 meters²). White and blue LED illumination was measured at the level of the rats’ eyes with a monochromatic light meter (Multi-Led TENMARS, Taiwan). Supplementary Figure S1 shows other details of the experimental system. All rats were euthanized at 08:00 AM in a CO₂ chamber. All experimental procedures were performed according to Ukrainian animal welfare regulations and were approved by the Poltava National Agricultural Academy Ethical Commission.