Encyclopedia of Experiments: Biology
A subscription to JoVE is required to view this content.
Transcript
Clean the abdominal surface with alcohol and povidone-iodine, and maintain sterile conditions throughout the operation. Perform a vertical incision for the laparotomy of pregnant dams, and cut all layers separately. Identify the uterine horns on each side and determine the candidate fetuses for the surgery.
Operate on two fetuses in each uterine horn if there is an even number of fetuses on each side, and on one fetus in each uterine horn if there is an odd number. Using 2x magnification glasses for visualization, position the uterine horn in a transverse fashion. Position the pups facing upward between two fingers using the eyes and the tail as a guide.
Apply gentle pressure to the pup's head to allow extension of the head, and visualization of the neck. Perform tracheal occlusion, or TO, using an atraumatic needle and a 6-0 polypropylene suture. Keep the placenta on the side and far from the entrance and exit points of the needle.
Insert the needle transversely through the side of the uterus, away from the placenta, through the one-third anterior part of the neck. Then, move the needle gently to the midline of the neck. Direct it to the anterior part, then, exit the neck between the trachea and opposite the carotid sheath and uterus.
Knot the suture, taking care to maintain the integrity of the membranes and uterine wall. Replace the uterine horn in the abdomen and inject 2 milliliters of warm, sterile saline into the peritoneal cavity before closure.
Place a running 5-0 polyglactin suture to close the abdominal wall, and close the skin with a non-running silk suture. Apply 0.1 mg/kg of buprenorphine intraperitoneally for analgesia and allow the dam to recover in a warm incubator. Observe that the operated animal can feed itself and keep it alone in its individual cage.
