The coupling between Ca²⁺ channels and the exocytotic Ca²⁺ sensor at hair cell ribbon synapses varies tonotopically along the mature cochlea

The cochlea processes auditory signals over a wide range of frequencies and intensities. However, the transfer characteristics at hair cell ribbon synapses are still poorly understood at different frequency locations along the cochlea. Using recordings from mature gerbils, we report here a surprisingly strong block of exocytosis by the slow Ca²⁺ buffer EGTA (10 mM) in basal hair cells tuned to high frequencies (~30 kHz). In addition, using recordings from gerbil, mouse, and bullfrog auditory organs, we find that the spatial coupling between Ca²⁺influx and exocytosis changes from nanodomain in low-frequency tuned hair cells (~<2 kHz) to progressively more microdomain in high-frequency cells (~>2 kHz). Hair cell synapses have thus developed remarkable frequency-dependent tuning of exocytosis: accurate low-latency encoding of onset and offset of sound intensity in the cochlea’s base and submillisecond encoding of membrane receptor potential fluctuations in the apex for precise phase-locking to sound signals. We also found that synaptic vesicle pool recovery from depletion was sensitive to high concentrations of EGTA, suggesting that intracellular Ca²⁺ buffers play an important role in vesicle recruitment in both low- and high-frequency hair cells. In conclusion, our results indicate that microdomain coupling is important for exocytosis in high-frequency hair cells, suggesting a novel hypothesis for why these cells are more susceptible to sound-induced damage than low-frequency cells; high-frequency inner hair cells must have a low Ca²⁺ buffer capacity to sustain exocytosis, thus making them more prone to Ca²⁺-induced cytotoxicity.