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Genetically modified mice targeting the vesicular acetylcholine transporter (VAChT) gene have been used as a tool to better understand the role of ACh signaling in specific regions of the brain (Prado et al., 2006; Guzman et al., 2011; de Castro et al., 2009). VAChT stores acetylcholine (ACh) in synaptic vesicles, and changes in this transporter expression directly interferes with ACh release. Here, we use a mouse line with VAChT deletion in the pedunculopontine tegmental nucleus (PPT) to evaluate the consequences of decreased cholinergic signaling from the PPT in vivo. Based on the efferent connectivity from the PPT, our hypothesis is that decreased cholinergic signaling from the PPT will affect specific behavioural outputs such as gait, motor control and sleep. Improving our understanding of cholinergic signaling from the PPT and how it relates to behavioural abnormalities may lead to the development of novel treatments for the symptoms of neurological diseases such as PD and PSP, which show degeneration of PPT neurons (Hirsch et al. 1987).