The segregation of Calb1, Calb2, and Prph neurons reveals distinct and mixed neuronal populations and projections to hair cells in the inner ear and central nuclei

BACKGROUND: Knockin mouse models expressing calbindin (Calb1), calretinin (Calb2), and peripherin (Prph) exhibit changes in hair cells (HCs), spiral ganglion neurons (SGN), vestibular ganglion neurons (VGNs), and their central projections. RESULTS: Developing cristae HCs show strong Calb1-positive expression, but adult HCs are mainly Calb2-positive. Utricle and saccule initially have Calb2-positive HCs and later develop Calb1-positive HCs in the striola region. Inner hair cells (IHCs) and outer hair cells (OHCs) in the cochlea express Calb2 early on. Calb1 expression in OHCs overlaps with Calb2; the expression of Myo7a, Calb1, and Calb2 reaches the apex later. SGNs and VGNs exhibit distinct Calb1 and Calb2 patterns but include a subpopulation with mixed expression. Central fibers are Calb1- and Calb2-positive early in the developing cochlear nuclei (CN) and vestibular nuclei (VN) but remain highly Prph-positive. VGNs innervate the lateral and VN, which are positive for Calb2 and Prph. Distinct Calb1-positive neurons overlap with the anterior (A) and ventral (V) cochlear nuclei (AVCN, PVCN) with Calb2, while the dorsal cochlear nucleus (DCN) shows segregation of Calb2 and Calb1. CONCLUSION: We offer insights into the timing of how neuronal identity and connectivity are regulated in the auditory and vestibular systems, as shown by the expression of Calb1, Calb2, and Prph.