Kaolin-Induced Hydrocephalus Model with Necroptosis Inhibitors

The hydrocephalus model was induced via kaolin injection into the cisterna magna.²² (link) The mice were anaesthetized with pentobarbital (40 mg/kg intraperitoneally). The skin was incised along the midline of the neck, the muscles were separated, and the cisterna magna was exposed under aseptic conditions. Then, a 30‐gauge needle (Hamilton, Switzerland), which was custom bent to 30°‐45°, was inserted into the cisterna magna. A 6‐μl volume of a sterile kaolin suspension (100 mg/mL, Sigma‐Aldrich) in 0.9% sterile saline was injected for approximately 30 seconds. The experimental groups were divided into the sham group, hydrocephalus group, hydrocephalus+Nec‐1 group (Nec‐1 group) and hydrocephalus+GSK872 group (GSK872 group). The specific necroptosis inhibitors Nec‐1 (Selleck) and GSK872 (Merck Millipore, Billerica, MA, USA) were dissolved in DMSO and diluted to a final concentration of 25 mM.²³ (link) A total of 1 μL of Nec‐1 and 1 μL of GSK872 were injected four times into the lateral ventricles (coordinates relative to the bregma: 0.5 mm posterior, 1 mm lateral, 2.2 mm deep) of the mice once per week after the kaolin injection.