Bda 3000

Rats received CT label 50-51 days after P5 CTX (n =5), P10 CTX (n=5), or Sham surgery at P5 (n = 3) or P10 (n = 2). Animals were anesthetized with a mixture of ketamine (73 mg/kg) and xylazine (7 mg/kg) given i.p., and the right tympanic bulla was accessed ventrally using blunt forceps. The CT was visualized in the tympanic bulla and cut at the point where it crosses the ossicles. The proximal end of the CT was carefully moved onto a shelf of bone and a small piece of cotton soaked in dimethyl sulfoxide (DMSO) was placed on the nerve for 15 s. Crystals of 3 kDa biotinylated dextran amine (BDA; Invitrogen, BDA-3000) were liberally applied onto the cut CT, and left for at least 15 min. The tympanic bulla was then filled with Kwik-Sil (World Precision Instruments) to keep the CT and dye in place. The wound was closed with 2-3 sutures. Animals were placed on a heating pad during recovery from the anesthetic. Once sternal recumbency was regained, animals were returned to their home cages.

The GL of rats treated with PLX5622 or Vehicle was labeled at P14, P25, P40 (n = 4–6/condition) or P68 (n = 3–5/condition). Animals were anesthetized with i.p. injections of Brevital Sodium (60 mg/kg) and a small incision was made on the ventral portion of the neck. The GL was located medial and ventral to the tympanic bulla, cut, and DMSO followed by biotinylated dextran amine (Invitrogen, BDA-3000) were applied to the nerve for 15 minutes. The incision was sutured, animals were monitored until sternal recumbency was observed, and then returned to their home cages. Animals were given 5 mg/kg carprofen i.p. after the surgery and 24 hours later.