Methods

1. Weigh the mouse to determine the dose of Avertin to administer (15-17 |L 100% Avertin/g body wt).

2. Draw up the appropriate volume into a sterile 1-mL disposable syringe. Exclude any air bubbles. (Have the needle of the syringe facing up, flick gently to dislodge any bubbles, and push the barrel of the piston until the bubbles are expelled.)

3. Restrain the animal, and introduce the needle into the abdomen of the mouse, avoiding both the diaphragm and the bladder. Withdraw the barrel of the syringe slightly—a small air bubble is observed if the needle has been inserted correctly into the intraperitoneal space. Inject the anesthetic and wait awhile before withdrawing the needle. Accidental subcutaneous injection is revealed by backleakage of drops of Avertin through the skin. The mouse will remain fully anesthetized for 0.5-1 h.

4. An adequate depth of anesthesia is indicated by an absence of a blinking reflex while blowing on the eyes and the maintenance of rapid breathing. Following surgery, the animal is observed to have regained mobility before being left to recover in a quiet, warm place.

1. Weigh the rat to determine the dose of CRC to administer (0.275 |L CRC/g body wt). Inject the rat intraperitoneally as described for the mouse (above). If the rat is too big to be restrained with one hand, use one of the many commercially available plastic animal restrainers or ask another person to assist in restraining the rat.

2. The rat will remain fully anesthetized for 20-30 min. An adequate depth of anesthesia is indicated by an absence of a jerk reflex on pinching the animal's paw and the maintenance of rapid breathing.

3. Following surgery, the animal is observed to have regained some mobility before being left to recover in a quiet, warm place.

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