Differences in mechano-transducer channel kinetics underlie tonotopic distribution of fast adaptation in auditory hair cells

The first step in audition is a deflection of the sensory hair bundle that opens mechanically gated channels, depolarizing the sensory hair cells. Two mechanism of adaptation of mechano-electric transducer (MET) channels have been identified in turtle auditory hair cells. The rate of fast adaptation varies tonotopically and is postulated to underlie a mechanical tuning mechanism in turtle auditory hair cells. Fast adaptation is driven by a calcium-dependent feedback process associated with MET channels. The purpose of this paper is to test the hypothesis that fast adaptation contributes to MET channel kinetics and that variations in channel kinetics underlie the tonotopic distribution of fast adaptation. To test for kinetic differences, the open channel blocker dihydrostreptomycin (DHS) was used. DHS blocked MET currents from low-frequency cells (IC50 = 14 +/- 2 muM) more effectively than high-frequency cells (IC50 = 75 +/- 5 muM), suggesting differences in MET channel properties. DHS block showed similar calcium sensitivities at both papilla locations. No difference in calcium permeation or block of the transducer channels was observed, indicating that the DHS effect was not due to differences in the channel pore. Slowing adaptation increased DHS efficacy, and speeding adaptation decreased DHS efficacy, suggesting that adaptation was influencing DHS block. DHS block of MET channels slowed adaptation, most likely by reducing the peak intraciliary calcium concentration achieved, supporting the hypothesis that the rate of adaptation varies with the calcium load per stereocilia. Another channel blocker, amiloride showed similar efficacy for high- and low-frequency cells with an IC50 of 24.2 +/- 0.5 muM and a Hill coefficient of 2 but appeared to block high- frequency channels faster than low-frequency channels. To further explore MET channel differences between papilla locations, stationary noise analysis was performed. Spectral analysis of the noise gave half power frequencies of 1,185 +/- 148 Hz (n = 6) and 551 +/- 145 Hz (n = 5) for high- and low-frequency cells in 2.8 mM external calcium. The half power frequency showed similar calcium sensitivity to that of adaptation shifting to 768 +/- 205 Hz (n = 4) and 289 +/- 63 Hz (n = 4) for high- and low- frequency cells in 0.25 mM external calcium. Both the pharmacological data and the noise analysis data are consistent with the hypothesis that the tonotopic distribution of fast adaptation is in part due to differences in MET channel kinetics. An increase in the number of MET channels per stereocilia (termed summation) and or intrinsic differences in MET channel kinetics may be the underlying mechanism involved in establishing the gradient.