Lysolecithin-induced demyelinating injury

Lysolecithin-induced demyelination was carried out in the ventrolateral spinal white matter of approximately 8-week-old mice. Anesthesia was induced and maintained by peritoneal injection of a mixture of ketamine (90 mg/kg) and xylazine (10 mg/kg). After exposing the spinal vertebrae at the level of T9-T12, meningeal tissue in the intervertebral space was cleared, and the dura was pierced with a dental needle. 0.5 μl of 1% lysolecithin (L-a-lysophosphatidylcholine, Sigma L4129) via a Hamilton syringe attached a glass micropipette was injected into the ventrolateral white matter via a stereotactic apparatus. Spinal cord tissues carrying the lesions were collected at different time points. For demyelination in the brain, focal demyelinating lesions were induced by stereotaxic injection of 1.5 μl lysolecithin solution (1% in 0.9% NaCl) into the corpus callosum at coordinates: 1 mm lateral, 1.3 mm rostral to bregma, 1.7 mm deep to brain surface) using a glass-capillary connected to a 10 μl Hamilton syringe. Animals were left to recover in a warm chamber before being returned into their housing cages. LPC-induced injuries were conducted in a genotype-blinded manner.