This study investigates the role of mutated alpha-synuclein protein in neuroinflammation using a transgenic mouse model. The research focuses on how alpha-synuclein fibrils promote aggregation and subsequent neural cell death.
Position a transgenic mouse in a dorsal position. The mouse expresses a mutated form of the alpha-synuclein protein, which is prone to aggregation.
Take a suspension of alpha-synuclein fibrils, aggregates of misfolded alpha-synuclein protein.
Inject the fibrils into the peritoneal cavity to allow their systemic distribution.
Once in the bloodstream, the fibrils reach the central nervous system and bind to endothelial cells of the blood-brain barrier.
The fibrils are transported via vesicles across the cells and enter the nervous tissue. They then invade neurons through endocytosis and promote aggregation of the mutated alpha-synuclein proteins.
These aggregates are released by exocytosis and spread to neighboring neurons, microglia, and astrocytes.
Recognition of the aggregates triggers microglia to release reactive oxygen species, which damages the tissue, and cytokines, which recruit additional immune cells. This results in neuroinflammation that leads to neural cell death.
Allow the mouse to recover to assess alpha-synuclein-mediated neuroinflammation.
For intraperitoneal injections, briefly narcotize the animals in an anesthesia chamber with isoflurane. Again, pinch the animal's toe and confirm that it does not withdraw its hindlimb to ensure proper anesthesia. Fill a 27-gauge disposable hypodermic syringe with 50 microliters of sonicated alpha-synuclein fibrils or PBS.
Hold the animal in a dorsal position, with the head facing away from the investigator, and downward at approximately 45 degrees. Directly inject into the peritoneum of the mouse without penetrating the small intestine or cecum located behind the abdominal wall. Monitor animals until they have recovered from the anesthesia.
繁體中文
