Intraperitoneal glucose tolerance test (IPGTT) 22/Jan/2018 RIKENMPP_031_001 test 1 1 testSt test modification

The glucose tolerance test measures the clearance of an intraperitoneally injected glucose load from the body. Animals are fasted for approximately 18 hours (overnight), before glucose solution is administered by intraperitoneal (IP) injection. Subsequently, the blood glucose level is measured at different time points during the following 2 hours.

These procedures are applicable to the mouse phenotyping platform at the RIKEN BRC. Testing should be performed by trained staff members who understand the safety and hygiene rules required by RIKEN. Any queries, comments, or suggestions, relating either to this SOP in general or to a specific problem encountered during a procedure, should be addressed to the Blood Tests Unit Leader.

Testing is carried out in accordance with common animal handling procedures. Coveralls (in the rearing facility) or laboratory coats (in the laboratory facility) must be worn with gloves at all times in the work area. If animals are dropped on the floor, treat them in accordance with the safety and hygiene guidelines of the rearing facility at RIKEN. If something unexpected happens during the primary screening, it must be quickly reported to the Blood Tests Unit Leader and recorded.

Any glucose values that overshoot the measurement limit (600 ml/dL) are shown as 601 ml/dL in the file.

Confirm precision of glucose meter according to the manufacturer’s instructions.

Glucose meter (GLUCOCARD PlusCare GT-1840; Arkray) The range of measureable values for this equipment is 10-600 ml/dL (point 5.1). Test sticks (G-sensor; Arkray) Scalpel scissors Balance Timer Clean cages

Glucose solution 20% (0.9 NaCl) Syringe (1 ml) Gauge needle (26G 1/2)

Fast mice for 16-18 hours (overnight). Use new cages and water bottles. On the following day, prepare the experiment record sheet, syringe, sticks for glucose measurement and glucose solution. Record the weight of each mouse. Calculate and record the volume of 20 % glucose solution required (2 g of glucose/kg body weight). Cut the tip of the tail with the scissors. Place a drop of blood (< 5 uL) on the glucose meter using the G-sensor set and record the baseline glucose level (= T0 result) in experiment record sheet. Inject the mouse intraperitoneally with the appropriate amount of glucose solution, as previously determined (point 9.4) and note the time-point of injection on the record sheet. Measure the glucose level in dropped blood from the tail of mouse at 15, 30, 60 and 120 minutes (= T15, T30, T60, and T120) after injection of the glucose solution. Result are recorded in the record sheet. At the end of the experiment, return the mice to their cages and make sure that plenty of water and food is available to the animals.

[2018.1.22] For v1.1, Glucose meter was changed to "GT-1840".