The brain was sectioned at 100-μm thicknesses using the vibratome (Leica VT 1200 Semi-Automatic Vibrating Blade Microtome, Leica Microsystems, Germany).
The sections were placed in 0.05 M Tris-buffered saline (Merck, Germany) and mounted onto gelatin-coated slides. Sections were air dried and incubated in 0.5% w/v cresyl violet stain (Sigma, USA). Sections were dehydrated and rehydrated in increasing concentrations of ethanol and xylene respectively with a cover slip and mounting medium. Stained sections were viewed under the microscope to identify microinjection sites of drug administration and electrode placements. Only data from animals with correct cannula implants (95% of the rats) were included in statistical analyses.
The sections were placed in 0.05 M Tris-buffered saline (Merck, Germany) and mounted onto gelatin-coated slides. Sections were air dried and incubated in 0.5% w/v cresyl violet stain (Sigma, USA). Sections were dehydrated and rehydrated in increasing concentrations of ethanol and xylene respectively with a cover slip and mounting medium. Stained sections were viewed under the microscope to identify microinjection sites of drug administration and electrode placements. Only data from animals with correct cannula implants (95% of the rats) were included in statistical analyses.