Pyruvate Tolerance Test
From Bridges Lab Protocols
Materials
- Glucometer - AccuChek Advantage
- Glucose Test Strips - AccuChek Comfort Curve. Order these through Materiels Service by calling 936-6077 and ordering product 2535. For information see [here]
- Scale
- Beaker for weighing mice
- Syringes
- 2.5g pyruvate in 10 mL PBS- This will correspond to 2.5 g/kg injections.
- Timer
Protocol
- Remove food from mice for about 6h.
- Typically starve the mice at 9AM and aim to start injections at 3 PM
- Weigh mice, mark tails if necessary and take fasting glucose measurement via a tail clip.
- Prepare pyruvate syringes with 10 uL per g mouse weight (ie for a 30g mouse, 300 uL).
- At approximately 1 min intervals, inject appropriate amount of pyruvate into interperitoneal cavity of the mouse.
- Immobilize mouse and restrain tail with one hand
- Aim needle between the midline and the hip bone
- Insert syringe (do not inject) into cavity
- Eject syringe.
- At desired intervals (normally 15, 30, 45, 60, 75, 90, 105 and 120 min), take blood glucose measurements from tail vein. If needed re-snip the tail vein. When measuring glucose just lift the tail of the mouse, while leaving it in the cage, rather than removing and restraining the mouse.
- Analyse data by both % change from fasting glucose and absolute values.
