F05312

Six-week-old male B6C3F1 hybrid were obtained from Harlan Laboratories (Madison, WI, USA) and housed under controlled pathogen-free conditions in accordance with the recommendations of the University of Wisconsin Institutional Animal Care and Use Committee. Mice were fed 87 kcal week⁻¹ of the control diet (F05312; Bio-Serv, Flemington, NJ, USA) and were individually housed with ad libitum access to water. This level of food intake is ~95% ad libitum for the B6C3F1 strain, so all food was consumed. Following two weeks of facility acclimation, mice were randomized into five treatment groups fed the control diet supplemented with increasing concentrations of dietary lithium carbonate (2 months old; n = 10/group): Group 1) 0.0 g/kg/day Li₂CO₃; Group 2) 0.6 g/kg/day Li₂CO₃; Group 3) 1.2 g/kg/day Li₂CO₃; Group 4) 1.8 g/kg/day Li₂CO₃; Group 5) 2.4 g/kg/day Li₂CO_3. Li2CO3-supplemented mice were administered an additional drinking bottle containing saline (0.45% NaCl) to offset polyuria, a common side effect of lithium treatment. Mice consumed dietary lithium for 4 months and were euthanized at 6 months of age. Mice were weighed every 1–2 weeks throughout the duration of the study. Body composition analysis was conducted using the Lunar PIXImus machine and software.
Brains were isolated, bisected, embedded in OCT, frozen in liquid nitrogen, and stored at −80°C until further processing.

Six-week-old male B6C3F1 hybrid were obtained from Harlan Laboratories (Madison, WI, USA) and housed under controlled pathogen-free conditions in accordance with the recommendations of the University of Wisconsin Institutional Animal Care and Use Committee. Mice were fed 87 kcal week⁻¹ of the control diet (F05312; Bio-Serv, Flemington, NJ, USA) and were individually housed with ad libitum access to water. This level of food intake is ~95% ad libitum for the B6C3F1 strain, so all food was consumed. Following two weeks of facility acclimation, mice were randomized into five treatment groups fed the control diet supplemented with increasing concentrations of dietary lithium carbonate (2 months old; n = 10/group): Group 1) 0.0 g/kg/day Li₂CO₃; Group 2) 0.6 g/kg/day Li₂CO₃; Group 3) 1.2 g/kg/day Li₂CO₃; Group 4) 1.8 g/kg/day Li₂CO₃; Group 5) 2.4 g/kg/day Li₂CO_3. Li2CO3-supplemented mice were administered an additional drinking bottle containing saline (0.45% NaCl) to offset polyuria, a common side effect of lithium treatment. Mice consumed dietary lithium for 4 months and were euthanized at 6 months of age. Mice were weighed every 1–2 weeks throughout the duration of the study. Body composition analysis was conducted using the Lunar PIXImus machine and software.
Brains were isolated, bisected, embedded in OCT, frozen in liquid nitrogen, and stored at −80°C until further processing.