Pyruvate Tolerance Test

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.