Chemogenetic silencing of cholinergic neurons impairs coordinated motor performance. A, NBM/SI basal forebrain cholinergic neuron transduction with AAV-DIO-hM4Di-mCherry in hemizygous ChAT-Cre mice. B, hM4Di-mCherry fusion protein expression in ChAT-positive neurons (scale bar: 200 μm). C, J60 delivered 30 min before behavior severely impaired performance on rotarod training in mice expressing hM4Di in NBM/SI cholinergic neurons (repeated-measures ANOVA, p = 0.0033, F(1,18) = 11.44). D, Rotarod latencies averaged for three trials each day (repeated measures ANOVA with post hoc Sidak's comparison test, *p < 0.05, **p < 0.01). Data presented as mean ± SEM.