Intracerebroventricular (i.c.v.) bolus injections were performed as previously described [65 (link)]. The ASO was dissolved and diluted in sterile D-PBS and sterilized by filtration through a 0.22-µm filter. The DMSXL mice were anesthetized with isoflurane (4% in O2, 1 L/min) and were placed in a rat stereotaxic frame (Stoelting, Wood Dale, IL, USA) with ultra-precise manipulator arms (Stoelting). The isoflurane was increased to 2% in O2 at a flow rate of 0.5 L/min (ABBOTT, Mississauga, ON, Canada) through a P28 mouse gas anesthesia head holder. A craniotomy was performed with a 2-mm ball head drill bit to inject 5 µL of the ASO with a Neuros syringe (Hamilton, City, QC, Canada) equipped with a 33-G blunt point needle at a rate of 1 µL/s at the following coordinates: X = 1.1 mm, Y = −0.2 mm, and Z = −2.2 mm from the bregma. After 5 min, the needle was slowly withdrawn, and the skin incision was sutured.
The dose-response experimental group sizes were: Saline (n = 5), 9 µg of ASO (n = 5), 19 µg of ASO (n = 5), 38 µg of ASO (n = 4), and 75 µg of ASO (n = 5). The time-effect experimental group sizes were 5 mice per time point for the IONIS-486178 ASO and 5 mice for the saline per time point.
The dose-response experimental group sizes were: Saline (n = 5), 9 µg of ASO (n = 5), 19 µg of ASO (n = 5), 38 µg of ASO (n = 4), and 75 µg of ASO (n = 5). The time-effect experimental group sizes were 5 mice per time point for the IONIS-486178 ASO and 5 mice for the saline per time point.