(A) Representative image of a control and ChAT-CKO mouse demonstrates severe kyphosis and unkempt coat. (B) ChAT-CKO mice exhibit significantly increased kyphotic curvature during locomotion (Mann-Whitney U = 35, p<0.0001). (C) ChAT-CKO mice exhibit a significantly reduced latency to fall during forelimb suspension (Mann-Whitney U = 71.5, p<0.0001). (D, E) ChAT-CKO mice are hypoactive in the open field (horizontal movement, t(23)=2.345, p=0.028; vertical rears, welch-corrected t(15.1) = 2.345, p=0.033). (F) Performance on the accelerated rotarod does not significantly differ from controls (two-way repeated measures ANOVA, genotype, F(1,43)=0.75, p=0.389; trial, F(9,387)=55.63, p<0.0001; interaction, F(9,387)=1.194, p=0.297). (G - I) ChAT-CKO mouse gait is abnormal during locomotion (paw angle, two-way ANOVA main effect of genotype, F(1,56)=30.54, p<0.0001, main effect of limb F(3,56)=51.02, p<0.0001, interaction F(3,56)=13.51, p<0.0001, post-hoc Sidak’s multiple comparisons test. Stance width, t(14)=3.329, p=0.005. Stride length, two-way ANOVA genotype F(1,28)=3.164, p=0.086, limb F(1,28)=0.02, p=0.887, interaction F(1,28)=0.0001, p=0.989).