Z033429

Adult (2–3 months) BALB/c mice obtained from our in-house breeding colony were used in all experiments. ICV cannulation was performed under aseptic conditions as described previously^{58 (link)}. In brief, 4 μg/μL polybrene was incubated at 37 °C for 15 min, and ICV injection of 5 μL mixture was performed for ≥20 min.
At day 3 after ICV injection, Motor Performance Test-Accelerating Rotating Rod Test was performed. The rotating rod apparatus was used to evaluate motor function. The mice were placed on the rod (3 cm in diameter) for four trials. Each trial lasted for a maximum of 5 min, during which the rotating rod underwent linear acceleration from 0 to 30 rpm and then remained at maximum speed. Animals were scored for their latency to fall (in seconds) in each trial.
Then the mice were deeply anesthetized and euthanized by transcardiac perfusion with fixative solution (4% paraformaldehyde and 2% glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.4, 50 mL/animal). The brains were harvested and postfixed in 4% paraformaldehyde, dehydrated in increasing concentrations of sucrose (30–60%), infiltrated in OCT Tissue Freezing Medium (380148, Leica), and plastic-embedded. Frozen sections (15 μm) were cut, mounted on coverslips, and then dried at room temperature and stained with NeuN (ab128886, Abcam, 1:500) and GFAP (Z033429, Dako, 1:1000).