Isopropanol 2 propanol

Dexamethasone, mifepristone, fomepizole, and common reagents were purchased from Sigma-Aldrich. Lipopolysaccharide (E. coli, serotype O111:B4 L2630) was from List Biological Laboratories (Campbell, CA) or from Sigma-Aldrich. Pure anhydrous ethyl alcohol or ethanol (200 proof/100%, Koptec), propanol (Sigma-Aldrich), and isopropanol (2-propanol, Sigma-Aldrich) were obtained commercially.

P1 and P7 Wistar rats were anaesthetised by overdose injection of Lethabarb before transcardial perfusion with 0.05% (w/v) heparin (David Bull Laboratories, Australia) in phosphate buffered saline (PBS), followed by 4% paraformaldehyde (w/v; Sigma-Aldrich, Australia) in 0.1 M Sorenson’s Buffer (4% PFA; pH 7.4). Eyes and whole brains were dissected out and postfixed in 4% PFA for 30 minutes before cryoprotection in diethylpyrocarbonate (DEPC; Sigma-Aldrich, Australia) treated PBS containing 30% sucrose (w/v; Sigma-Aldrich, Australia) overnight at 4°C. PBS/sucrose solution was gradually replaced with Jung tissue freezing medium (Leica Microsystems, Australia) over several days. After infusion tissues were snap frozen in isopropanol (2-propanol; Sigma-Aldrich, Australia). 20 μM sections were cut on a cryostat, placed on SuperFrost Plus slides (Menzel-Gläser, Germany), and stored at −80°C until processing by in situ hybridisation (ISH).