Difference between revisions of "Pyruvate Tolerance Test"

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(Updated layout of the protocol)
m (fixed typo)
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* Remove food from mice for about 6h.  
 
* Remove food from mice for about 6h.  
 
* Typically starve the mice at 9AM and aim to start injections at 3 PM
 
* 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.
+
* 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).
 
* 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.
 
* At approximately 1 min intervals, inject appropriate amount of pyruvate into interperitoneal cavity of the mouse.

Revision as of 16:39, 27 April 2017

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.