Dumont 5 45 forceps

Facial nerve injuries were performed as described (Olmstead et al., 2015 (link)) with some modifications. Briefly, adult animals were anesthetized by isofluorane inhalation, as described above, and after shaving the right ear posterior region, a surgical 5 mm skin incision was performed to expose the facial nerve branches. The most dorsal branch of the facial nerve innervating the LAL muscles was carefully cleared avoiding direct manipulation. In the NMJ degeneration protocol, a section of 4 mm of the facial nerve branch was transected; in turn, for the reinnervation protocol, the facial nerve branch was crushed for 30 s using Dumont #5/45 forceps (Fine Science Tools). Control experiments only considered skin incision and facial nerve exposure. Finally, the skin was sutured using absorbable monofilament surgical suture (Ethicon Vicryl USP 6–0) and animals were monitored until their recovery.

Two facial nerve injury protocols were performed [23 (link)] leading to either chronic denervation or NMJ reinnervation. Briefly, male and female adult mice (3–6 months old) were anesthetized by isoflurane inhalation (2.5% v/v isoflurane with a 0.8-1 L/min oxygen mixture). After shaving the right ear posterior region, a surgical 5-mm skin incision was performed to expose the facial nerve branches. The posterior auricular branch of the facial nerve, which innervates the LAL muscle, was carefully cleared avoiding direct manipulation. For the chronic denervation protocol, a 5-mm portion of the facial nerve branch was transected. For the reinnervation protocol, the facial nerve branch was crushed during 30 s using Dumont #5/45 forceps (Fine Science Tools). Finally, the skin was sutured using absorbable monofilament surgical suture (Ethicon Vicryl USP 6-0), and animals were monitored until their recovery. Control experiments only considered skin incision and facial nerve exposure.