Dense cholinergic projections to auditory and multisensory nuclei of the intercollicular midbrain

Cholinergic axons from the pedunculopontine tegmental nucleus (PPT) innervate the inferior colliculus where they are positioned to modulate both excitatory and inhibitory circuits across the central nucleus and adjacent cortical regions. More rostral regions of the auditory midbrain include the nucleus of the brachium of the inferior colliculus (NBIC), the intercollicular tegmentum (ICt) and the rostral pole of the inferior colliculus (ICrp). These regions appear especially important for multisensory integration and contribute to orienting behavior and many aspects of auditory perception. These regions appear to receive cholinergic innervation but little is known about the distribution of cholinergic axons in these regions or the cells that they contact. The present study used immunostaining to examine the distribution of cholinergic axons and then used chemically-specific viral tracing to examine cholinergic projections from the PPT to the intercollicular areas in male and female transgenic rats. Staining with antibodies against vesicular acetylcholine transporter revealed dense cholinergic innervation throughout the NBIC, ICt and ICrp. Deposits of viral vector into the PPT labeled cholinergic axons bilaterally in the NBIC, ICt and ICrp. In each area, the projections were denser on the ipsilateral side. The axons appeared morphologically similar across the three areas. In each area, en passant and terminal boutons from these axons appeared in the neuropil and also in close apposition to cell bodies. Immunostaining with a marker for GABAergic cells suggested that the cholinergic axons likely contact both GABAergic and non-GABAergic cells in the NBIC, ICt and ICrp. Thus, the cholinergic axons could affect multisensory processing by modulating excitatory and inhibitory circuits in the NBIC, ICt and ICrp. The similarity of axons and their targets suggests there may be a common function for cholinergic innervation across the three areas. Given what is known about the PPT, such functions could be associated with arousal, sleep-wake cycle, reward and plasticity.