Disorganized central projections of Neurod1CKO inner ear neurons show aberrant interconnections between the auditory and vestibular systems. (A,B) Lipophilic differently colored NeuroVue dyes were applied to the apex and base to label cochlear afferents. In the lateral view of the cochlear nucleus of the control, dye tracing shows the normal segregated basal and apical cochlear afferents of the auditory nerve and segregated projections to the AVCN and DCN. In contrast, reduced, disorganized, and overlapping central projections are shown in the cochlear nucleus of Neurod1CKO mice. Note unusual fibers from the apex projecting away from the cochlear nucleus and auditory nerve (arrowhead). (C) Coronal sections of control show the segregation of basal and apical afferents. (D,D’) Sections of Neurod1CKO show an overlap of disorganized fibers from the apex and base. Dotted lines in corresponding figures indicate the section planes in (A,B). (E,F) Dye tracing from the utricle and the posterior canal crista (PCC) shows vestibular afferents innervating the lateral, medial, and superior vestibular nuclei that are immediately adjacent to the AVCN in the control mice. A single dye tracing from the utricle shows vestibular afferents reach vestibular nuclei and form a profound projection to the AVCN in Neurod1CKO mice. The area of the AVCN is indicated by dotted circle. Scale bars: 100 µm. AN, auditory nerve; AVCN, anteroventral cochlear nucleus; DCN, dorsal cochlear nucleus; LVN, MVN, SVN, lateral, medial, and superior vestibular nuclei; VN, vestibular nerve.