Standard ain93g rodent diet

We purchased 61 male BALB/c mice (age 10 weeks) from Animal Resources Centre, Canning Vale, WA, Australia. The BALB/c mice were housed in groups of 4, in OptiMICE cages (Animal Care Systems, CO, USA), with corn cob bedding (Sanichips, Harlan Laboratories, USA) at the Animal Facility, Queensland Brain Institute, The University of Queensland, Australia. The animal housing conditions were maintained with a 12-h light-dark cycle. All of the animals had free access to water and food. The animals were given either a control diet (Standard AIN93G Rodent diet with 1500 IU vitamin D3/kg, Specialty Feeds, WA, Australia) or a vitamin D-deficient diet (Vitamin D Deficient AIN93G Rodent diet, Specialty Feeds, WA, Australia) (Supplementary table S1). Following behavioural testing, a separate group of AVD-deficient BALB/c mice (n = 6) were returned to the control diet (supplemented with a vitamin D containing diet) for 10 weeks. See Figure 1 for an outline of the timing for the diets used in these experiments. The experimental work was completed with approval from the University of Queensland Animal Ethics Committee (QBI/376/15/NHMRC), under the guidelines of the National Health and Medical Research Council of Australia.

Upon delivery, all animals were allowed to acclimatize for at least 1 week before commencement of dietary manipulation. Rats were randomly assigned to either a control diet (CON, Standard AIN93G rodent diet, 6% total fat including 1.05% total saturated fatty acids; Specialty Feeds, Perth, Australia) or WD (SF00-219, 21% total fat including 1.80% total saturated fats and 0.15% cholesterol; Specialty Feeds, Perth, Australia) and remained on this diet for 12 weeks.

A total of 101 BALB/c mice (71 male and 30 female) were used in this study. Ten-week old BALB/c mice (Animal Resources Centre, Canning Vale, WA, Australia) were obtained and housed in groups of four animals in individually ventilated OptiMICE cages, with bedding and nesting material at the Queensland Brain Institute Animal House Facility, The University of Queensland, Australia.
The mice were assigned to either a control diet (Standard AIN93G Rodent diet with 1,500 IU vitamin D₃/kg (prior to irradiation with 25 kGy), Specialty Feeds, WA, Australia) or a vitamin D-deficient diet (Vitamin D Deficient AIN93G Rodent diet with 0 I.U vitamin D₃/kg irradiated with 25 kGy, Specialty Feeds, WA, Australia) for 10 weeks prior to the start of behavioural testing and for the entire duration of the experimental procedures. The mice were maintained on a 12-hour light-dark cycle (lights on at 07:00 h) with ad libitum access to food and water. They were housed under incandescent lighting free from UVB radiation. All experimental work was performed with approval from The University of Queensland Animal Ethics Committee, under the guidelines of the National Health and Medical Research Council of Australia.