C3h hej

Mice were obtained from Jackson Laboratory and bred in the facilities of the NeuroBsik consortium (VU University Amsterdam, The Netherlands or Harlan Laboratories, Horst, The Netherlands; 129S1/SvImJ n = 61, A/J n = 49, BALB/cJ n = 47, C3H/HeJ n = 29, C57BL/6J n = 112, DBA/2J n = 40, FVB/NJ n = 49, NOD/ShiLtJ n = 46) or subjected to experiments 2 weeks after shipment from Jackson laboratories to the testing facility (WSB/EiJ n = 14, PWK/PhJ n = 15 and CAST/EiJ n = 14). Male 8 to 12 week old mice were singly housed on sawdust in standard Makrolon type II cages enriched with cardboard nesting material for at least one week prior to experiments, with water and food ad libitum (7:00/19:00 lights on/off; providing an abrupt phase transition). We only used male mice to avoid possible impact of estrous cycle on longitudinal behavioral assessments. Experiments were carried out in accordance with the European Communities Council Directive of 24 November 1986 (86/609/EEC), and with approval of the local animal care and use committee of the VU University Amsterdam, The Netherlands.

All procedures were approved by the Bavarian Animal Research Authority, Animal Research Authority of Schleswig-Holstein, and University of Pittsburgh, PA. Frozen and paraffin embedded tissues were procured to carry out experiments at SRM University, India according to the Institutional Animal Ethics Committee (IAEC) permission. Mouse strains C3H/HeJ (#000659) and JF1/MsJ (#003720) were purchased from Jackson laboratory (Bar Harbor, ME, USA) and housed in specific pathogen free conditions. Food and water were available ad libitum. Three developmental stages, namely, embryonic day (E) 18 (completion of embryonic lung development), postnatal day (P) 28 (completion of bulk alveolar formation) and P70 (completion of lung growth and maturity) were selected for the microarray studies. E18 embryos (days post coitum) were stage matched. Mice were anesthetized (4 mg/kg xylene and 188 mg/kg ketamine i.p.) and the posterior aorta was severed. To obtain tissue for mRNA and protein analysis (n = 5 mice/strain/stage, female), the diaphragm was punctured, and the chest cavity opened. Lungs were excised, frozen in liquid nitrogen, and stored (−80 °C). Whole lung was used for RNA extraction in case of stage E18. The left lobe of the lung was used for RNA extraction and the right lobe was used for protein extraction for P28 and P70.

A total of 23 mouse strains (129X1/SvJ, A/J, AKR/J, B10.S-H2s/SgMcdJ (B10.S), BALB/cJ, BPL/1J, BPN/3J, C3H/HeJ, C57BL/6J, C57BL/10J, CBA/J, CZECHII/EiJ, DBA/1J, DBA/2J, FVB/NJ, JF1/MsJ, MOLF/EiJ, MRL/MpJ, NOD/ShiLtJ, NU/J, PWD/PhJ, PWK/PhJ, SJL/J and SWR/J were purchased from the Jackson Laboratory (Bar Harbor, ME). All mice, including B10.S-Histh^SJL and B10.S-Histh^SJL ISRC lines, were generated and maintained under specific pathogen-free conditions in the vivarium of the Given Medical Building at the University of Vermont according to National Institutes of Health guidelines. All animal studies were approved by the Institutional Animal Care and Use Committee of the University of Vermont.

Eight-week-old male mice, C57BL/6N from Charles River (Wilmington, MA) and C57BL/6J and C3H/HeJ mice from Jackson Labs (Bar Harbor, ME), were obtained (n = 30 mice/strain) for these studies. Animals were acclimated for 7 days and then randomly assigned to a control AIN-93M (10% kcals from fat) or a HF (45% kcals from fat; Harlan Teklad, TD.06415) diet for 24 wk. Body weight and food intake were recorded throughout the study. Total feed efficiency was calculated by determining the gain in body weight (mg) per energy unit consumed (kcal) [35 (link)]. Venous tail blood was collected following a 6 hr fast for evaluation of glucose and insulin at 4 wk intervals. After 24 wk, mice were anesthetized (ketamine/xylazine cocktail 70 and 30 mg/kg body weight, resp.) as previously reported and whole body DXA (Lunar PIXI, GE Medical Systems, Madison, WI) scans were performed. Mice were exsanguinated via the carotid artery. An aliquot of blood was collected for total white blood cell (WBC) counts and the remainder processed for plasma in EDTA coated tubes and stored at −80°C. All procedures were approved by the Institutional Animal Care and Use Committee of Oklahoma State University.