Objective

The objective of this experiment was to address: 1) whether normal efferent system function is required for normal cochlear tuning as measured by distortion product otoacoustic emission (DPOAE) suppression in humans and 2) whether cochlear function, assessed by DPOAE suppression tuning, is normal in a small group of patients with auditory neuropathy.

Design

DPOAE suppression tuning curves (STCs) are similar to other physiologic measures of tuning. They are generated by evoking a DPOAE with two simultaneously presented pure tones and then suppressing the distortion product with a third tone of varying frequency and level. In this study, DPOAE STCs were generated with f2 frequencies of 1500, 3000, and 6000 Hz in 15 normal-hearing adults and four subjects with documented auditory neuropathy. Tuning curve width, slope and tip characteristics, as well as rate of suppression growth were measured in each group. Contralateral suppression of otoacoustic emissions (OAEs) was also recorded as an index of medial efferent function.

Results

Results show that the four subjects with auditory neuropathy lacked efferent suppression of OAEs. However, these four subjects showed normal estimates of cochlear tuning as measured by DPOAE suppression results.

Conclusions

This finding suggests that normal efferent system function is not required at the time of test for normal DPOAE suppression tuning. It also suggests that cochlear function as evaluated by detailed measures of DPOAE suppression, is normal in these "typical" patients with auditory neuropathy.

Neuropilin-1 (Nrp1) encodes the transmembrane cellular receptor neuropilin-1, which is associated with cardiovascular and neuronal development and was within the peak SNP interval on chromosome 8 in our prior GWAS study on age-related hearing loss (ARHL) in mice. In this study, we generated and characterized an inner ear-specific Nrp1 conditional knockout (CKO) mouse line because Nrp1 constitutive knockouts are embryonic lethal. In situ hybridization demonstrated weak Nrp1 mRNA expression late in embryonic cochlear development, but increased expression in early postnatal stages when cochlear hair cell innervation patterns have been shown to mature. At postnatal day 5, Nrp1 CKO mice showed disorganized outer spiral bundles and enlarged microvessels of the stria vascularis (SV) but normal spiral ganglion cell (SGN) density and presynaptic ribbon body counts; however, we observed enlarged SV microvessels, reduced SGN density, and a reduction of presynaptic ribbons in the outer hair cell region of 4-month-old Nrp1 CKO mice. In addition, we demonstrated elevated hearing thresholds of the 2-month-old and 4-month-old Nrp1 CKO mice at frequencies ranging from 4 to 32kHz when compared to 2-month-old mice. These data suggest that conditional loss of Nrp1 in the inner ear leads to progressive hearing loss in mice. We also demonstrated that mice with a truncated variant of Nrp1 show cochlear axon guidance defects and that exogenous semaphorin-3A, a known neuropilin-1 receptor agonist, repels SGN axons in vitro. These data suggest that Neuropilin-1/Semaphorin-3A signaling may also serve a role in neuronal pathfinding in the developing cochlea. In summary, our results here support a model whereby Neuropilin-1/Semaphorin-3A signaling is critical for the functional and morphological integrity of the cochlea and that Nrp1 may play a role in ARHL.