design: Young male ChAT-ZsGreen transgenic mice were killed by transcardiac perfusion with 4% formaldehyde. Following a 5-h postfixation in this fixative, the brains were infiltrated overnight with sterile 20% sucrose solution in sterile PBS. The cholinergic neurons were immunostained with our novel ImmunoLCM method. Cholinergic neurons were isolated via laser-capture microdissection from the caudate putamen (CPU), and subjected to RNA-sequencing (Illumina).