Three-week-old C57BL/6J female mice were purchased from The Jackson Laboratory (Bar Harbor, ME) and were randomly assigned to either experimental (treatment) or control groups and housed 5 mice per cage. All animals were maintained in the UCSF animal care facility, with a light cycle of 12 h. (light (7 am-7 pm) and 12 h. dark (7 pm-next day 7 am)) and were fed a standard rodent diet. The animal care facility is a barrier facility, accredited by Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). All procedures are conducted under the approval of UCSF Animal Care and Use Committee, and animals are checked daily for signs of distress and discomfort, with necessary care administered by a veterinarian. Water was given via plastic water bottles to avoid a loss of fluoride through absorption onto glass surfaces. Control mice were given deionized water and treatment groups were given deionized drinking water supplemented with 50 mg/L sodium fluoride (50ppmF, Sigma-Aldrich, St. Louis, MO) beginning at 21 days of age, and continuing for up to 42 weeks. The volume of water consumed was measured once per week. One group of 30 mice (group 1), including 15 control and 15 fluoride treated mice were maintained for 7 weeks and then tested for anxiety related behaviors using the open field and elevated plus maze. A second group of 40 mice (20 control and 20 fluoride treated), were also tested in the open field and elevated plus maze after 7 weeks, retested at 23 weeks, and maintained for up to 42 weeks. At 12 week and 42 weeks brain fluoride was measured in fluoride treated and control mice. 

Note: Only Group 2 data at the 23 week time point for the elevated plus maze (Figure 3) were extracted. Duplicate testing in Group 1 animals for the 7 week time point found similar results for this endpoint.

Three-week-old C57BL/6J female mice were purchased from The Jackson Laboratory (Bar Harbor, ME) and were randomly assigned to either experimental (treatment) or control groups and housed 5 mice per cage. All animals were maintained in the UCSF animal care facility, with a light cycle of 12 h. (light (7 am-7 pm) and 12 h. dark (7 pm-next day 7 am)) and were fed a standard rodent diet. The animal care facility is a barrier facility, accredited by Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). All procedures are conducted under the approval of UCSF Animal Care and Use Committee, and animals are checked daily for signs of distress and discomfort, with necessary care administered by a veterinarian. Water was given via plastic water bottles to avoid a loss of fluoride through absorption onto glass surfaces. Control mice were given deionized water and treatment groups were given deionized drinking water supplemented with 50 mg/L sodium fluoride (50ppmF, Sigma-Aldrich, St. Louis, MO) beginning at 21 days of age, and continuing for up to 42 weeks. The volume of water consumed was measured once per week. One group of 30 mice (group 1), including 15 control and 15 fluoride treated mice were maintained for 7 weeks and then tested for anxiety related behaviors using the open field and elevated plus maze. A second group of 40 mice (20 control and 20 fluoride treated), were also tested in the open field and elevated plus maze after 7 weeks, retested at 23 weeks, and maintained for up to 42 weeks. At 12 week and 42 weeks brain fluoride was measured in fluoride treated and control mice. 

Note: Only Group 1 data were extracted since duplicate testing in Group 2 animals found similar results.