After the completion of behavioral tests on transgenic mice, animals were euthanized with CO2 and decapitated. Brains were rapidly removed and cut in half through the midline. Half of the brain was placed in 4% paraformaldehyde for 24 hours to post-fix for GFP imaging and then stored in phosphate buffer. The other half of the brain was dissected into forebrain, midbrain, and cerebellum sections, and snap-frozen in liquid nitrogen for subsequent RNA isolation using Trizol® (Invitrogen) and qPCR. To image GFP fluorescence, 40-μm sagittal brain sections were cut using a vibrating blade microtome (Leica Microsystems, Bannockburn, IL) and mounted on slides. Images were acquired using a Zeiss confocal microscope, and visualized using Zeiss LSM software (Carl Zeiss MicroImaging, LLC, Thornwood, NY).
