The dorsal cochlear nucleus (DCN) is the first relay of the central auditory system and is involved in the localization of sound in the vertical plane. The DCN is also the site of integration of other sensory inputs. Therefore knowledge of synaptic inputs to the DCN is of importance to understand possible interactions between the auditory and other sensory circuits in this structure. We recently developed a method that allows delivery of fluorescent dyes within precisely localised tissue areas (Barker et al., 2008). Using this system, in excised brain tissue from decapitated rats, we were able to deliver dextran amine rhodamine conjugates to the Lister Hooded rat DCN and trace retrograde projections coming from the lateral vestibular nucleus. Dextran amine conjugates were also delivered to the lateral vestibular nucleus and synaptic terminals could be localised within the DCN confirming anterograde projections from the lateral vestibular nucleus to the DCN. Vestibulo-cochlear projections were characterised within the sagittal and the coronal plane. The projections were mainly within the deep and fusiform cell layers of the DCN and fibre tracts could only be discriminated in the sagittal plane. Therefore to assess functional synaptic projections from the lateral vestibular nucleus to the DCN, whole cell recordings were performed in sagittal slices. Stimulating the lateral vestibular nucleus elicited glutamatergic synaptic currents in fusiform and granule cells that were blocked by glutamate receptor antagonists NBQX and D-AP5. Vesicular glutamate transporters (VGLUT’s) selectively package glutamate into synaptic vesicles. By combining the dextran amine neuronal tracing method with immunocytochemistry, we were able to show projections from the lateral vestibular nucleus predominantly colabeled with VGLUT2 in the DCN, whereas very few colabeled with VGLUT1. In conclusion, we have shown that the lateral vestibular nucleus sends functional glutamatergic projections into the DCN, mainly mediated by the VGLU2 vesicular transporter. This suggests functional links between the auditory and the vestibular system and ultimately that balance and locomotion could interact with the sound localization in the vertical plane.