Intracerebroventricular Delivery of AAV Vectors

Intracerebroventricular injections of AAV were performed as previously described with some minor modifications [7 (link), 28 ]. Briefly, a 32-gauge needle (product #7803–04, 0.5 in. custom length, point style 4, 12 degrees, Hamilton Company) attached to a 10 μl syringe (Hamilton Company) was inserted into the lateral ventricle of cryoanesthetized C57BL/6J pups at postnatal day 0. The needle was inserted at a 30-degree angle from the surface of the head, and held at a depth of approximately two millimeters. Two microliters (1.5E10 genomes/μl) of AAV2/9-(G₄C₂)₂ or AAV2/9-(G₄C₂)₁₄₉ solution was manually injected into each lateral ventricle. Following injection, pups were placed on a heated pad until they recovered from anesthesia, at which time they were placed back into their home cage.