This article details the procedure for spinal cord extraction in a euthanized mouse, emphasizing the importance of maintaining sterility throughout the process. The steps outlined ensure the integrity of the spinal cord for subsequent analysis.
For spinal cord extraction, begin with a euthanized mouse in the prone position. Spray ethanol to prevent any potential contamination from fur.
Cut through the midline from the head to the tail and open the skin to reveal the underlying tissue. Examine the mouse’s skull to locate the craniocervical junction that connects the skull to the cervical spine.
Next, incise the craniocervical junction perpendicularly to sever the connection between the head and neck. Dissect the spine till the fifth lumbar vertebra. Subsequently, cut the spine at the fifth lumbar vertebra to sever the connection from the lower vertebral levels.
Now, turn the mouse to the supine position. Slit the peritoneum and the chest wall to expose both the abdominal and thoracic cavity. Remove the peritoneal and retroperitoneal organs to access the vertebral column.
Cut the ribs to separate the vertebral column from the rest of the body. Last, rinse the isolated spinal cord with a cold saline solution to wash away any blood contaminants.
After euthanizing the animal in a CO2 chamber, soak the fur with 70% ethanol and allow it to dry. Pin the mouse in a prone position and then make a midline incision extending craniocaudally from the posterior neck to the lumbar spine.
Next, use forceps to raise the dermal flaps bilaterally and expose the subcutaneous tissue. Palpate the mouse skull until the craniocervical junction is identified. Use scissors angled perpendicular to the animal to cut through the cervical muscles and spine at the craniocervical junction. Then, dissect the spine caudally and use scissors to perform a complete caudal transection at the fifth lumbar vertebrae.
Next, position the mouse in a supine position and, after making an incision along the midline from the neck to the abdomen, retract the skin laterally and then cut open the peritoneum.
Cranially, open the chest wall with scissors. After removing the peritoneal and retroperitoneal organs, use forceps and a surgical blade to cut the ribs, leaving approximately 5 millimeters of ribs protruding from the vertebral column. Remove the vertebral column from the rest of the body and wash twice with cold PBS.
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