ChI cells in NAc are successfully infected utilizing the rabies monosynaptic tracing method. (A) Injection schedule and types of viruses used for axonal tracing. (B) Schematic diagram showing the areas of the brain injected with the viruses. (C) Representative image of the cell in the NAc infected with the mixture of helper viruses (red, mCherry) and immunolabeled with the anti-ChI antibody. Scale bar, 10 μm. (D) Series of low magnification images of the sagittal sections of the mouse line ChAT-CRE crossed with the ChI-L10a-EGFP injected with the helper (mCherry) virus into the NAc. Sections were labeled with the mCherry (red) and GFP (green) antibodies to enable visualization of the infection efficacy in the ChI cells. The majority of infected cells, double-labeled with mCherry and GFP (yellow) are found in the NAc, with no major infection of the ChIs in the adjacent areas of the dorsal striatum, basal forebrain, and septum. Distance from the bregma: ∼1.54 mm (left image); ∼0.74 mm (middle image); ∼0.38 mm. Scale bars, 500 μm.