Td nmr minispec live mice analyzer

Mgll^−/− mice were generated as described previously [20 (link)]. Mice were backcrossed onto C57BL/6J for at least 10 generations. Mice were bred and maintained on a regular light dark cycle (14 h light, 10 h dark) at 22 ± 1 °C in a barrier facility in specific pathogen free quality. Animals had ad libitum access to chow diet (12 kJ% fat, V1126, Ssniff Spezialdiaeten GmbH, Soest, Germany) or were fed HFD (45kJ% fat, E15744, Ssniff Spezialdiaeten GmbH, Soest, Germany). HFD-treatment started at 6 weeks of age for 22 weeks. Body mass composition was determined by TD-NMR miniSpec Live Mice Analyzer (Bruker Optics, Billerica, MA, USA). Stool samples were collected three times from all mice, at 6 weeks of age (6 weeks old), after 8 weeks of HFD (14 weeks old) and after 22 weeks of HFD (28 weeks old). All the samples were stored at −80 °C until further use. All animal experiments were approved by the Austrian Federal Ministry for Science, Research, and Economy (protocol number BMBWF-66.007/0030-V/3b/2018) and the ethics committee of the University of Graz, and were conducted in compliance with the council of Europe Convention (ETS 123).

Body composition was determined by NMR spectroscopy using a TD-NMR miniSpec Live Mice Analyzer (Bruker Optics). For metabolic phenotyping, mice were single-housed in a laboratory animal monitoring system (PhenoMaster, TSE Systems) with ad libitum access to HFD and water. Food intake, locomotor activity, and oxygen consumption rate were determined for light and dark phase on four consecutive days. Body temperature was measured using a rectal probe RET-3 (Physitemp). For glucose tolerance test, mice were injected intraperitoneally (ip) with 1.6 g per kg body weight following a 6 h fast. For insulin tolerance test, mice were injected ip with 0.75 IU insulin per kg body weight following a 4 h fast. Blood glucose was determined by Wellion CALLA glucometer (Med Trust). Plasma lipid parameters were determined using following commercial kits: FA (NEFA-HR2, Wako Diagnostics), TAG (Infinity triglycerides, Thermo Fisher Scientific), glycerol (free glycerol reagent, Sigma-Aldrich), total cholesterol (cholesterol CHOD-PAP kit; Roche Applied Science), and ketone bodies (β-Hydroxybutyrate Assay Kit, Cayman Chemical). Plasma insulin levels were determined using Ultra-Sensitive Mouse Insulin ELISA Kit (Crystal Chem). Plasma ANP levels were determined using Atrial Natriuretic Peptide EIA Kit (Sigma-Aldrich). Plasma FGF21 levels were determined using Rat/Mouse FGF21 ELISA Kit (Sigma-Aldrich).