The World's Most Bizarre Ku-0059436 History

RosaNICD/NICD mice were crossed with Adipoq-Cre mice and the male offspring RosaNICD/-::AdipoqCre were crossed with female RosaNICD/NICD to get the desired phenotype of RosaNICD/NICD::AdipoqCre (Sitaxentan Ad-NICD mice). Male Ad-NICD mice were mated with female RosaNICD/NICD to generate the male mice used in the experiments. RosaNICD/NICD, used as controls, and RosaNICD/NICD::AdipoqCre (Ad-NICD) were both in the albino C57BL/6J background. An experimental cohort of male mice was sacrificed at 1 month of age and another experimental cohort was sacrificed at 3 months of age. The 1-month old cohort included 8 control and 9 Ad-NICD male mice. The 3-month old cohort included 7 control and 6 Ad-NICD male mice. Another cohort (5 control and 8 Ad-NICD mice) was used to evaluate the growth curves of the mice starting from 22 days old until the age of 90 days. The genotyping primers as well as a more detailed description of the RosaNICD/NICD::AdipoqCre model can be found in the supplementary material. The mice were housed at 22?��C, 50% humidity with a 12?h light/dark cycle with ad libitum access to water and food (Prolab Isopro RMH 3000 5P76 irradiated diet, LabDiet, St Louis, MO). 2.2. Body fat composition and metabolic tests in mice Body fat composition was assessed using NMR-MRI-based technology (EchoMRI, Houston, TX). Blood glucose was measured using a Precision Xtra glucose meter (Abbott, Chicago, IL) in blood collected from the tails of mice. Glucose tolerance was assessed by injecting intraperitoneally a single dose of d-glucose (1?g/kg) after a 16?h overnight fasting with free access to water. Insulin tolerance was assessed by injecting mice intraperitoneally with 0.75?units/kg insulin (Humalog, Eli Lilly, Indianapolis, IN) after a 4?h fasting with free access to water. 2.3. Serum metabolites Blood was drawn under isoflurane anesthesia with cardiac puncture and was allowed to clot at room temperature for 30?min. Serum was collected after centrifuging the blood at 2000?��?g for 30?min at 4?��C. Triglycerides and cholesterol concentrations were measured using a colorimetric and a fluorometric assay kit, respectively, from Cayman Chemical (Ann Arbor, MI). Liver triglycerides were also quantified using the same kit. Concentrations of non-esterified fatty acids were measured photometrically using the NEFA-HR kit (Wako Chemicals, Japan). Leptin and insulin were measured using ELISA kits from R&D systems (Minneapolis, MN) and Mercodia (Sweden), respectively. 2.4. Histology Tissue was immersed in 10% formalin or in O.C.T. (Sakura, Torrance, CA) for the frozen sections. Hematoxylin and Eosin (H&E) stained sections were prepared by the Department of Pathology, University of Pittsburgh.