Hypervulnerability to Sound Exposure through Impaired Adaptive Proliferation of Peroxisomes

A deficiency in pejvakin, a protein of unknown function, causes a strikingly heterogeneous form of human deafness. Pejvakin-deficient (Pjvk(-/-)) mice also exhibit variable auditory phenotypes. Correlation between their hearing thresholds and the number of pups per cage suggest a possible harmful effect of pup vocalizations. Direct sound or electrical stimulation show that the cochlear sensory hair cells and auditory pathway neurons of Pjvk(-/-) mice and patients are exceptionally vulnerable to sound. Subcellular analysis revealed that pejvakin is associated with peroxisomes and required for their oxidative-stress-induced proliferation. Pjvk(-/-) cochleas display features of marked oxidative stress and impaired antioxidant defenses, and peroxisomes in Pjvk(-/-) hair cells show structural abnormalities after the onset of hearing. Noise exposure rapidly upregulates Pjvk cochlear transcription in wild-type mice and triggers peroxisome proliferation in hair cells and primary auditory neurons. Our results reveal that the antioxidant activity of peroxisomes protects the auditory system against noise-induced damage

Atypical Tuning and Amplification Mechanisms in Gecko Auditory Hair Cells

The auditory papilla of geckos contains two zones of sensory hair cells, one covered by a continuous tectorial membrane affixed to the hair bundles and the other by discrete tectorial sallets each surmounting a transverse row of bundles. Gecko papillae are thought to encode sound frequencies up to 5 kHz, but little is known about the hair cell electrical properties or their role in frequency tuning. We recorded from hair cells in the isolated auditory papilla of the crested gecko, Correlophus ciliatus, and found that in both the nonsalletal region and part of the salletal region, the cells displayed electrical tuning organized tonotopically. Along the salletal zone, occupying the apical two-thirds of the papilla, hair bundle length decreased threefold and stereociliary complement increased 1.5-fold. The two morphological variations predict a 13-fold gradient in bundle stiffness, confirmed experimentally, which, when coupled with salletal mass, could provide passive mechanical resonances from 1 to 6 kHz. Sinusoidal electrical currents injected across the papilla evoked hair bundle oscillations at twice the stimulation frequency, consistent with fast electromechanical responses from hair bundles of two opposing orientations across the papilla. Evoked bundle oscillations were diminished by reducing Ca2+ influx, but not by blocking the mechanotransduction channels or inhibiting prestin action, thereby distinguishing them from known electromechanical mechanisms in hair cells. We suggest the phenomenon may be a manifestation of an electromechanical amplification that augments the passive mechanical tuning of the sallets over the high-frequency region

Calcium- and Otoferlin-Dependent Exocytosis by Immature Outer Hair Cells

International audienceImmature cochlear outer hair cells (OHCs) make transient synaptic contacts (ribbon synapses) with type I afferent nerve fibers, but direct evidence of synaptic vesicle exocytosis is still missing. We thus investigated calcium-dependent exocytosis in murine OHCs at postnatal day 2 (P2)–P3, a developmental stage when calcium current maximum amplitude was the highest. By using time-resolved patch-clamp capacitance measurements, we show that voltage step activation of L-type calcium channels triggers fast membrane capacitance increase. Capacitance increase displayed two kinetic components, which are likely to reflect two functionally distinct pools of synaptic vesicles, a readily releasable pool (RRP; τ = 79 ms) and a slowly releasable pool (τ = 870 ms). The RRP size and maximal release rate were estimated at ∼1200 vesicles and ∼15,000 vesicles/s, respectively. In addition, we found a linear relationship between capacitance increase and calcium influx, like in mature inner hair cells (IHCs). These results give strong support to the existence of efficient calcium-dependent neurotransmitter release in immature OHCs. Moreover, we show that immature OHCs, just like immature IHCs, are able to produce regenerative calcium-dependent action potentials that could trigger synaptic exocytosis in vivo . Finally, the evoked membrane capacitance increases were abolished in P2–P3 OHCs from mutant Otof −/− mice defective for otoferlin, despite normal calcium currents. We conclude that otoferlin, the putative major calcium sensor at IHC ribbon synapses, is essential to synaptic exocytosis in immature OHCs too

Control of exocytosis by synaptotagmins and otoferlin in auditory hair cells.

International audienceIn pre-hearing mice, vesicle exocytosis at cochlear inner hair cell (IHC) ribbon synapses is triggered by spontaneous Ca(2+) spikes. At the onset of hearing, IHC exocytosis is then exclusively driven by graded potentials, and is characterized by higher Ca(2+) efficiency and improved synchronization of vesicular release. The molecular players involved in this transition are still unknown. Here we addressed the involvement of synaptotagmins and otoferlin as putative Ca(2+) sensors in IHC exocytosis during postnatal maturation of the cochlea. Using cell capacitance measurements, we showed that Ca(2+)-evoked exocytosis in mouse IHCs switches from an otoferlin-independent to an otoferlin-dependent mechanism at postnatal day 4. During this early exocytotic period, several synaptotagmins (Syts), including Syt1, Syt2 and Syt7, were detected in IHCs. The exocytotic response as well as the release of the readily releasable vesicle pool (RRP) was, however, unchanged in newborn mutant mice lacking Syt1, Syt2 or Syt7 (Syt1(-/-), Syt2(-/-) and Syt7(-/-) mice). We only found a defect in RRP recovery in Syt1(-/-) mice which was apparent as a strongly reduced response to repetitive stimulations. In post-hearing Syt2(-/-) and Syt7(-/-) mutant mice, IHC synaptic exocytosis was unaffected. The transient expression of Syt1 and Syt2, which were no longer detected in IHCs after the onset of hearing, indicates that these two most common Ca(2+)-sensors in CNS synapses are not involved in mature IHCs. We suggest that otoferlin underlies highly efficient Ca(2+)-dependent membrane-membrane fusion, a process likely essential to increase the probability and synchrony of vesicle fusion events at the mature IHC ribbon synapse