2.1. Animal Care

All mice (C57BL/6J background strain, The Jackson Laboratory, Bar Harbor, ME) were singly-housed in conventional style open cages at the Florida State University (FSU) vivarium in accordance with institutional requirements for animal care. Mice were maintained on a reverse 12/12 hour dark/light cycle so that behavioral phenotyping could take place in the dark phase. Rodent chow (5001, Purina, Richmond VA) and water were provided ad libitum (unless otherwise specified). Experiments began at adulthood, defined as 4 months of age, and were only performed on male animals. Although sex-specific effects have been reported for rats and humans [57, 59, 80, 81], such has not been found for mice [16] so it was not necessary to utilize both sexes. Because mice also demonstrate a lack of diet-induced obesity (DIO) and associated prediabetic state in females [36], with the intent of applying our study for future application for diabetes, it was pragmatic to hone our current study to male mice. Taking the aforementioned into consideration, we acknowledge the potential study limitations due to the lack of females. All animal experiments were approved by the FSU Institutional Animal Care and Use Committee (IACUC) under protocol #1427 and were conducted in accordance with the American Veterinary Medicine Association (AVMA) and the National Institutes of Health (NIH). For final collection of tissues for biochemistry, mice were sacrificed by cervical dislocation (AVMA Guidelines on Euthanasia, June 2007). Use of the ARRIVE guidelines for reporting animal research was followed in the design of our manuscript [29]