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. 2021 May 19;41(20):4378–4391. doi: 10.1523/JNEUROSCI.2537-20.2021

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Copyright © 2021 Beurg et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license, which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Figure 5. — Ionic properties of MET channels from apical OHCs of Tmc1 p.T416K/T416K; Tmc2−/−. A, Three examples of single MET channel currents for Tmc1+/+; Tmc2−/−, ensemble average of 30 presentations (middle) and amplitude histogram (bottom) giving −7.0 pA at –84 mV. C and O indicate closed and open states of channel. B, Examples of single MET channel currents for Tmc1 p.T416K/T416K; Tmc2−/−, ensemble average of 100 presentations (middle) and amplitude histogram (bottom) giving −7.0 pA at −84 mV. C, Determination of MET channel Ca²⁺ permeability in isotonic external Ca²⁺ and internal Cs⁺ (see Materials and Methods). During a sinusoidal bundle vibration (ΔX, ±200 nm), the membrane potential (V_m) was swept from −30 to +80 mV and the MET current reversed polarity at arrowed potential. Reversal potential in Tmc1+/+; Tmc2−/− is ∼7 mV positive to that in Tmc1 p.T416K/T416K; Tmc2−/−. D, Collected reversal potentials and relative permeability P_Ca/P_Cs in six cells mean +/− SD. Control and mutant are significantly different (t test, ***p < 0.001). E, MET currents for bundle vibrations (ΔX, ±200 nm) in normal saline (black traces) and in presence of 100 μm DHS in Tmc1+/+; Tmc2−/− (top) and Tmc1 p.T416K/T416K; Tmc2−/− (bottom). F, Mean ± SD (N = 5) of MET current block by DHS in Tmc1+/+ and Tmc1 p.T416K/T416K. Hill plots give K_I = 15 μm (all points) and 66.0 μm, respectively, with Hill coefficients = 1 for both genotypes. For control curve, open symbols determined using bundle stimulation with a fluid et (K_I = 14.0 μm), whereas crosses obtained with a stiff glass probe (K_I = 15.4 μm). The similar K_I values for the different jstimulation methods rules out artifacts because of dilution of DHS by saline in the fluid jet.