Intravagal AAV injection in mice
Ayse Ulusoy, Rita Pinto-Costa, Angela Rollar, Donato Di Monte, Michael Klinkenberg, Michael Helwig, Shirley Lee, Sinead O'Sullivan
Abstract
The protocol details AAV vector injections into the mouse vagus nerve and describes pre and post-operative mouse handling.
Steps
Anesthetize the mouse with isoflurane and use pain killers (e.g. Temgesic) following the ethical committee procedures
Clean the surgery area with skin disinfectant, make a small incision at the midline of the neck, and
isolate the left vagus nerve. Place a holder underneath to keep it isolated and to form a base to insert the needle.
Using a micropump and a 35-gauge blunt steel needle fitted to a 10-ul NanoFil syringe, inject 800 nl AAV-containing solution. Keep the needle in place for another minute to avoid backflow.
Suture and clean the wound and return the mouse to a clean cage. Follow the ethical committee's recommendations for post-operative care.
post-operative drug treatments
Post-operative drug treatments:
To stimulate DREADDs, inject 1 mg/kg CNO (Tocris) dissolved in 0.9% saline intraperitoneally.
For in situ oxidative stress assessments dissolve DHE (Dihydroethidium) in saline/dimethyl sulfoxide (DMSO) (1:1 ratio) and inject intraperitoneally at a dose of 15 mg/kg.
