GMC11 project protocol

Glucose tolerance test in 6 inbred founder strains of the Collaborative Cross   (2020)

German Mouse Clinic and Department of Infection Genetics, HZI
With: Hrabě de Angelis M, Fuchs H, Gailus-Durner V, Rathkolb B, Wolf E, Lengger C, Kollmus H, Schughart K




German Mouse Clinic: Phenotyping Pipeline

Intraperitoneal glucose tolerance test (IPGTT): 13-14 wks of age

 

  • Project protocol - Contents

    Workflow and sampling

    Step
    Procedure
    		 Equipment
    Data collected
    1
    		 Mice are weighed Balance
    Body weight before fast
    2 Mice are fasted overnight (~16 h) - -
    3 Mice are weighed Balance Body weight after fast
    4 Post-fast glucose levels are measured prior to glucose injection Glucose meter Blood glucose at t=0
    5 Mice are injected (i.p.) with glucose solution - -
    6

    Glucose levels are measured at four timepoints within a 2-h period

    		 Glucose meter Blood glucose at t=15, 30, 60, 120 min

    Equipment and supplies

    • Balance: Ohaus (AV213C)
    • Restraining device
    • Syringes, needles
    • Scalpel blades
    • Glucose meter: Roche Diagnostic (Accu-Chek Aviva)
    • Glucose meter test strips: Roche Diagnostic
    • Timer

    Reagents and solutions

    • 20% glucose solution

    Procedure: Intraperitoneal glucose tolerance test (IPGTT)

      1. Mice are weighed and fasted overnight for ~16 h in a clean cage with access to water at all times.
      2. Mice are weighed again post-fasting in order to calculate the volume of 20% glucose solution required to give 2 g glucose/kg body weight.
      3. Each mouse is restrained in the restraining device with the tail exposed. The tip of the tail is scored using a scalpel blade and the first drop of blood is discarded.
      4. The second drop of blood (<5 µL) is placed on the test strip and fasting-levels of glucose (t=0) are measured by the glucose meter and recorded.
      5. The mouse is removed from the restraining device and injected intraperitoneally (i.p.) with the appropriate amount of glucose as determined above.
      6. Blood glucose levels are measured and recorded at 15, 30, 60, and 120 min post-injection using the technique described above using a fresh test strip.
      7. Light pressure is applied to the incision after each measurement to ensure minimal blood loss.
      8. After testing, mice are placed in a clean cage with water and food available ad libitum.

    Data collected by investigator

    • Body weight before and after fast
    • Blood glucose levels at t = 0, 15, 30, 60, 120 min post-injection
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