Cb6f1 j mice

Six-week-old CB6F1/J mice (Jackson Laboratory, Bar Harbor, ME, USA) were maintained for breeding 6- to 8- day-old (P6-P8) offspring with ad libitum access to food and water at a 12-h light and 12-h dark cycle in the animal facility of the Division of Life Sciences at the Nelson Biology Laboratories of Rutgers University. Offspring of either sex was used for the preparation of brain homogenate and of primary cerebellar granule cell culture. The Institutional Animal Care and Use Committee of Rutgers University approved all animal experiments (protocol no. 09-051).

Age-matched (6-10 weeks) female CB6F1/J mice (Jackson Laboratories, Bar Harbor, ME, USA) were used in all experiments. Mice were housed in the animal care facility at the National Institute of Allergy and Infectious Diseases under specific-pathogen-free conditions and maintained on standard rodent chow and water supplied ad libitum. All studies were reviewed and approved by the National Institutes of Health Animal Care and Use Committee.

Mice utilized for diet-induced obesity studies were male CB6F1/J mice (Jackson Laboratories Stock #100007) which are the F1 generation of a cross between male C57BL/6 and female Balb/c. Mice were obtained at three weeks of age, housed at Vanderbilt for three days on the standard chow diet after which time they were fed either control (10% calories from fat; 4.3% fat by weight, D12450Bi, Research Diets, New Brunswick, NJ), HFD (45% calories from fat; 24% fat by weight, D12451i, Research Diets), or very high fat diet (VHFD) (60% calories from fat; 34.9% fat by weight, D12492i, Research Diets). Mice were maintained on a 12-hour light/dark cycle and housed with three to five animals per cage on corncob bedding except during fasting when placed on Pure-o’Cel™ bedding (The Andersons, Maumee, OH). Mice were weighed weekly and body composition was assessed every four weeks by pulsed NMR (Bruker Instruments, The Woodlands, TX) at the Vanderbilt Mouse Metabolic Phenotyping Center. After 17 weeks of defined diet feeding, mice were injected via SC with 20 mg/kg DG-041 or vehicle twice daily for one week. At the termination of the experiment, mice were euthanized by isoflurane overdose and liver, epididymal fat pads, and skeletal muscle were dissected. All procedures were approved by the Institutional Animal Care and Use Committee at Vanderbilt University.

Adult (6–10 weeks old) female CB6F1/J mice or OT-I mice were purchased (The Jackson Laboratories, Bar Harbor, ME) or bred in-house. CB6F1/J neonates (6–7 days old) were obtained through timed breeding of BALB/c (female) and C57BL/6 (male) mice (The Jackson Laboratories).^{26 (link)} Mice were bred and housed under specific pathogen-free (SPF) conditions at the Uniformed Services University (USU). Studies were approved by and adhered to the USU Institutional Animal Care and Use Committee (IACUC).