Prolab isopro rmh 3000

Eight-week-old male C57BL/6J mice (Jackson Laboratories, Bar Harbor, ME) were used for behavioral pharmacology experiments. To visualize Pdyn-expressing neurons, we generated a Pdyn-GFP reporter line by crossing preprodynorphin-IRES-Cre mice (Crowley et al., 2016 (link); Bloodgood et al., 2020 (link); Al-Hasani et al., 2015 (link)) (Pdyn^IRES-Cre, B6;129S-Pdyn^{tm1.1(cre)Mjkr}/LowlJ, Jackson Laboratories Stock # 027958) and Rosa26-flox-stop-L10a-EGFP reporter mice (EGFP-L10a; B6;129S4-Gt(ROSA)26Sor^{tm9(EGFP/Rpl10a)Amc}/J, Jackson Laboratories Stock # 024750). For conditional knockout of BNST Pdyn, we used the Pdyn^lox/lox mouse line (Bloodgood et al., 2020 (link)). These mice were bred in the UNC facilities. All mice were group-housed for at least 3 days before being singly housed in polycarbonate cages (GM500, Tecniplast, Italy) with a 12:12 hr reversed dark-light cycle with lights off at 7:00am. Mice had unrestricted access to food (Prolab Isopro RMH 3000, LabDiet, St. Louis, MO) and H₂O. The UNC School of Medicine Institutional Animal Care and Use Committee approved all experiments. Procedures were conducted in accordance with the NIH Guidelines for the Care and Use of Laboratory Animals.

All animal experiments were conducted in accordance with the Animal Welfare Act and other federal statutes and regulations relating to animals and experiments involving animals and adhered to principles stated in the Guide for the Care and Use of Laboratory Animals (NRC Publication 2011 edition) using an Institutional Animal Care and Use Committee approved protocol. Male Sprague Dawley rats, 9–10 weeks old weighing 300–350 g (Charles River Laboratories, Wilmington, MA), were housed at 20–22°C (12 h light/dark cycle). Rats were given free access to nutritious rat chow (Prolab IsoPro RMH 3,000 from LabDiet, St. Louis, MO) and water ad libitum till 1 month after the blast exposure, when they reached a body weight of 400–450 g. We restricted diet for all rats including sham controls after 1 month so that the weight of the rats was maintained between 450 and 500 g until the completion of the study (1 year). Body weights were recorded 3 days a week and adjustments were made in the quantity of diet to maintain body weights within this range since these animals were also used for neurobehavioral functional tests summarized in a recent publication (18 (link)).

Mice in all experiments were fed ad libitum a diet of standard mouse chow ProLab IsoPro RMH 3000 (Lab Diet; St Louis MO).

The fetuses used in this study were generated from other ongoing projects within the lab in accordance with National Institutes of Health guidelines using methods and protocols approved by the Institutional Animal Care and Use Committee of the University of North Carolina (15–290).
C57BL6/J female mice (Jackson Labs, Bar Harbor, ME) were housed in groups of five or fewer, and breeder C57BL6/J males were single housed, in polycarbonate cages (39 x 20 x 16 cm) under controlled temperature and humidity with cob bedding and cotton nesting material, with free access to water and standard chow (Prolab Isopro RMH 3000, LabDiet, St. Louis, MO). Beginning about 4 hours into the light portion of a 12:12 light/dark cycle, 1–2 female mice (weighing at least 20g) were placed into a male’s cage for 1–2 hours. Females were then removed from the male’s cage and checked for the presence of a vaginal plug. GD 0 was defined as the beginning of the breeding period if a plug was found. Thus, 24 and 36 hours after the beginning of the breeding period would be GD 1 and 1.5, respectively.

Animal studies were performed in accordance with protocols approved by the Institutional Animal Care and Use Committee at the Texas A&M Health Science Center, Institute of Biosciences and Technology. Mice were fed with standard ProLab IsoPro RMH3000 (LabDiet).

The GK non-obese rat with type 2 diabetes mellitus (T2DM) and Wistar (Control) control animals were purchased from Charles River Laboratories (Wilmington, MA, United States). GK rats manifest spontaneous skeletal muscle and hepatic insulin resistance, mild hyperglycemia and normal lipidemia by 4 weeks of age. Six male GK and six Wistar controls were housed in pairs in the Animal Resource Center facilities of Case Western Reserve University with 12:12-h light-dark cycle and were fed standard diet chow (Prolab Isopro RMH 3000, LabDiet, St. Louis, MO, United States) ad libitum. The sample size was used to detect difference between bioenergetics parameters in rat skeletal muscle fibers (Lopes Martins et al., 2018 (link)). Male animals were selected to avoid any hormonal effects on energy metabolism during the menstrual cycle. Animals were euthanized at 28 weeks of age. The euthanasia was performed 7–8 a.m. on the day of study while food was available. All procedures were approved by Case Western Reserve University Institutional Animal Care and Use Committee and performed in accordance with the National Research Council guidelines for care and use of laboratory animals in research. Plasma insulin and glucose, and the bioenergetics of skeletal muscle mitochondria in these animals were previously published by our group (Lai et al., 2017 (link)).