Ilium xylazil 100

Mice were anesthetized with an intraperitoneal injection of 40 mg/kg Ketamine (Ceva Animal Health Pty Ltd, Amersham, UK) and 5 mg/kg Ilium Xylazil-100 (Troy Laboratories, New South Wales, Australia) or subcutaneously with a mixture of Ketamine (85 mg/kg; Parke-Davis, Berlin, Germany) and xylazine (4 mg/kg; Bayer AG, Leverkusen, Germany). Single intravitreal injections of 0.5 uL were administered at PN (Postnatal day) 14, with either 100 nM of Trichostatin A (TSA; Sigma) or 10 µM of GSK-J4 (Selleckchem, Texas, USA) administered in one eye and the contralateral eye receiving 0.0001% DMSO (diluted in 0.9% sodium chloride solution [Sigma]). GSK-J4 contained 0.0001% DMSO and was further diluted in 0.9% sodium chloride solution to make a 100 µM solution, while TSA (Trichostatin A) also contained 0.0001% DMSO with dilution to make a 100 nM solution. Assuming a 5 uL free vitreous volume [26 ], the final concentration of treatment was 10 nM TSA or 10 µM of GSK-J4. Anesthesia was reversed with Ilium Atipamezole (1 mg/kg; Troy Laboratories) or Antisedan (Atipamezole; 2 mg/kg; Orion Corp, Espoo, Finland).

Traumatic brain injury was induced by a controlled cortical impactor (CCI) TBI 0310 (Precision Systems and Instrumentation, LLC, Fairfax, VA, USA). TBI induction was described in our previous study [11 (link)]. Briefly, mice were anesthetized by intraperitoneal injection of Zoletil 50 (tiletamine hydrochloride and zolazepam hydrochloride, 25 mg/ml/kg, VIRBAC Laboratories, Carros, France) and Ilium Xylazil-100 (Xylazine, 10 mg/ml/kg, Troy Laboratories, Australia) before surgery. After shaving and disinfection, the mouse skull underwent craniotomy at the left cortex using a drill. A 3 mm diameter impact head was settled 0.5 mm posterior to the bregma and 0.5 mm lateral to the midline. The weight drops a head on the flat exposed brain surface with 2 mm impact depth, 5 m/s impact velocity, and 500 ms dwell time. After cortical brain injury, the mouse skull was replaced, and the skin was enclosed with wound clips. The animals were returned to their home cage under a heat lamp until recovery from anesthesia.