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Postural instability and gait disturbances, common disabilities in the elderly and frequently present in Parkinson's disease (PD), have been suggested to be related to dysfunctional cholinergic signaling in the brainstem. We investigated how longterm loss of cholinergic signaling from mesopontine nuclei influence motor behaviors. We selectively eliminated the vesicular acetylcholine transporter (VAChT) in pedunculopontine and laterodorsal tegmental nuclei cholinergic neurons to generate mice with selective mesopontine cholinergic deficiency (VAChT^{En1Creflox/flox}). VAChT^{En1Creflox/flox} mice did not show any gross health or neuromuscular abnormality on metabolic cages, wirehang and gripforce tests. Young VAChT^{En1Creflox/flox} mice (2–5 monthsold) presented motor learning/coordination deficits on the rotarod; moved slower, and had smaller steps on the catwalk, but showed no difference in …