To begin, transfer the prepared anesthetized mouse to the surgical area and isolate the surgical site with an adhesive drape. Under deep anesthesia, use a scalpel blade to make a ventral midline incision in the skin. Next, use scissors to separate the skin from the muscle layer.
With the help of a scalpel blade, make a smaller incision in the muscle layer. Once the bowels are visualized, use blunt forceps to locate the cecum and exteriorize it. Then ligate the cecum with a sterile 5/0 polyglactin absorbable suture.
Using a 27-gauge needle, perforate the secum through and through allowing the fecal content to leak. Gently squeeze the secum to externalize the fecal content with caution. Afterward, relocate the secum into the abdominal cavity and close the muscle layer with a sterile 5/0 absorbable suture.
Then use a 5/0 nylon non-absorbable suture to close the skin. After that, provide sterile saline via subcutaneous injection in the back of the animal. After four days of recovery from CLP, or sham surgery, use foam tape to attach the anesthetized mouse tail to a short metal chain.
Then attach the metal chain to a hook connected to a crossbar along the center of the cage. Affix a second small bar that can move along the crossbar to allow the animal greater movement ability. Then adjust the height of the suspended limbs to prevent the paws from contacting the chow pellets.
Ensure that the animals can move via their forelimbs using the metal grid on the cage floor. CLP animals demonstrated increased immune cell populations in their peritoneal lavage compared to sham animals. CLP and HLS lead to approximately 11.8%of body mass loss.
A marked decrease in soleus mass was observed when HLS was superimposed with CLP, indicating that smaller fibers comprise a major part of the muscle section. Compared to the control animals, CLP-HLS caused significant impairment in both the absolute and peak forces of the soleus.
