Mouse Resources Portal

Indirect Calorimetry (week 12, male mice prioritised) [Mice were fed on High Fat Diet (21.4% crude fat content, Western RD, 829100, Special Diets Services) from 4 weeks of age]: Respiratory metabolic function is assessed using the LabMaster system (TSE-systems). Mice are housed individually in calorimetry cages for a period of approximately 21 hours including a 5 hour light phase acclimatisation period. This is followed consecutively by a 12 hour dark (19:30 - 07:30) and minimum 3 hour light phase. Cumulative food intake, activity (beam breaks), volume of oxygen consumed and volume of carbon dioxide produced are all measured. From this data the respiratory exchange ratio and energy expenditure are derived. Water bottles are weighed before and after the experiment to assess water consumption.

ipGTT (week 13) [Mice were fed on High Fat Diet (21.4% crude fat content, Western RD, 829100, Special Diets Services) from 4 weeks of age]: Mice are fasted overnight (maximum of 16 hours) before the glucose tolerance test. A fasting blood sample is taken before a bolus of glucose is administered by intra-peritoneal injection. Blood samples are tested for glucose concentration (Accu-Check Aviva, Roche) at 15, 30, 60 and 120 minutes following the glucose administration, using strips modified such that there is no interference with maltose. Data are presented as plasma glucose concentration.

ipGTT (week 13) [Mice were fed on Mouse Breeder Diet (5021, Labdiet) from weaning]: Mice are fasted overnight (maximum of 18 hours) before the glucose tolerance test. A fasting blood sample is taken before a bolus of glucose is administered by intra-peritoneal injection. Blood samples are tested for glucose concentration (Accu-Check Aviva, Roche) at 15, 30, 60 and 120 minutes following the glucose administration, using strips modified such that there is no interference with maltose. Data are presented as plasma glucose concentration.

Core Temperature and Stress Induced Hyperthermia (15 weeks) [Mice were fed on High Fat Diet (21.4% crude fat content, Western RD, 829100, Special Diets Services) from 4 weeks of age]: Basal core body temperature is measured rectally using a TH-5 thermometer with a RET-3 probe (Viking Medical). The mice are thereafter placed in a clean cage and a second reading is taken after 15-30 minutes.

Plasma Chemistry Panel (week 16) [Mice were fed on High Fat Diet (21.4% crude fat content, Western RD, 829100, Special Diets Services) from 4 weeks of age]: Non-fasted mice are terminally anaesthetised with ketamine/xylazine, and blood is collected into Lithium/Heparin coated tubes via the retro-orbital sinus. Plasma is analysed for the following parameters on an Olympus AU400:
Electrolytes: Sodium, potassium, chloride

Comprehensive Clinical Chemistry Panel (week 16) [Mice were fed on Mouse Breeder Diet (5021, Labdiet) from weaning]: Non-fasted mice are terminally anaesthetised with ketamine/xylazine, and blood is collected into Lithium/Heparin coated tubes via the retro-orbital sinus. Plasma is analysed for the following parameters on an Olympus AU400:
Electrolytes: Sodium, potassium, chloride
Non-Fasted Metabolic Panel: Glucose, fructosamine, triglycerides, cholesterol, high density lipoprotein, low density lipoprotein, non-esterified free fatty acids (NEFAC) and glycerol
Pancreatic Enzyme: Amylase
Liver/Muscle Panel: Alanine aminotransferase, alkaline phosphatase, creatine kinase, aspartate aminotransferase, total bilirubin
Protein Parameters: Total protein, albumin
Kidney Panel: Creatinine, urea
Minerals and Iron: Calcium, magnesium, iron, phosphate For a subset of genes, plasma lactate dehydrogenase and uric acid are analysed.
Plasma insulin concentration is measured by Mesoscale Discovery (MSD) array technology.