7013 nih 31

All studies were conducted using male wild-type C57BL/6J mice purchased from Jackson Laboratories or bred in-house (Bar Harbor, ME). Mice were kept on a 14-hour light/10-hour dark schedule and housed 3-5 mice per cage until 1 week prior to study initiation, at which point animals were individually housed. We conducted studies in lean chow fed mice (7013 NIH-31, Teklad WI, 3.1 kcal/g, 18% kcal from fat, 59% kcal from carbohydrate, 23% kcal from protein) at 12-16 weeks of age. Studies in diet-induced obese mice dosed intraperitoneally with GABA transaminase inhibitors, mice treated with ethanolamine-O-sulfate in their drinking water, and mice treated with GABA-T targeted or scramble control antisense oligonucleotides (ASO) were performed after 8-10 weeks on a high fat diet (TD 06414, Teklad WI, 5.1 kcal/g, 60.3% kcal from fat, 21.3% kcal from carbohydrate, 18.4% kcal from protein; 20-26 weeks of age). For ASO studies, mice were stratified by body weight and assigned to an injection treatment (control or GABA-T). Studies in obese vagotomy and sham mice were performed after 9 weeks of high fat diet feeding. Unless fasted, mice had ad libitum access to food and water. All studies were approved by The University of Arizona Institutional Animal Care and Use Committee.