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      Prestin-Driven Cochlear Amplification Is Not Limited by the Outer Hair Cell Membrane Time Constant

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          Summary

          Outer hair cells (OHCs) provide amplification in the mammalian cochlea using somatic force generation underpinned by voltage-dependent conformational changes of the motor protein prestin. However, prestin must be gated by changes in membrane potential on a cycle-by-cycle basis and the periodic component of the receptor potential may be greatly attenuated by low-pass filtering due to the OHC time constant (τ m), questioning the functional relevance of this mechanism. Here, we measured τ m from OHCs with a range of characteristic frequencies (CF) and found that, at physiological endolymphatic calcium concentrations, approximately half of the mechanotransducer (MT) channels are opened at rest, depolarizing the membrane potential to near −40 mV. The depolarized resting potential activates a voltage-dependent K + conductance, thus minimizing τ m and expanding the membrane filter so there is little receptor potential attenuation at the cell's CF. These data suggest that minimal τ m filtering in vivo ensures optimal activation of prestin.

          Highlights

          ► Cochlear amplification via prestin limited by outer hair cell (OHC) time constant ► Endolymph Ca 2+ opens half the transduction channels, depolarizing OHC to −40 mV ► Depolarization activates K + conductance, reduces time constant and expands filter ► Minimal membrane filtering in vivo ensures prestin activation over all frequencies

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          Most cited references51

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          Prestin is the motor protein of cochlear outer hair cells.

          The outer and inner hair cells of the mammalian cochlea perform different functions. In response to changes in membrane potential, the cylindrical outer hair cell rapidly alters its length and stiffness. These mechanical changes, driven by putative molecular motors, are assumed to produce amplification of vibrations in the cochlea that are transduced by inner hair cells. Here we have identified an abundant complementary DNA from a gene, designated Prestin, which is specifically expressed in outer hair cells. Regions of the encoded protein show moderate sequence similarity to pendrin and related sulphate/anion transport proteins. Voltage-induced shape changes can be elicited in cultured human kidney cells that express prestin. The mechanical response of outer hair cells to voltage change is accompanied by a 'gating current', which is manifested as nonlinear capacitance. We also demonstrate this nonlinear capacitance in transfected kidney cells. We conclude that prestin is the motor protein of the cochlear outer hair cell.
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            Prestin is required for electromotility of the outer hair cell and for the cochlear amplifier.

            Hearing sensitivity in mammals is enhanced by more than 40 dB (that is, 100-fold) by mechanical amplification thought to be generated by one class of cochlear sensory cells, the outer hair cells. In addition to the mechano-electrical transduction required for auditory sensation, mammalian outer hair cells also perform electromechanical transduction, whereby transmembrane voltage drives cellular length changes at audio frequencies in vitro. This electromotility is thought to arise through voltage-gated conformational changes in a membrane protein, and prestin has been proposed as this molecular motor. Here we show that targeted deletion of prestin in mice results in loss of outer hair cell electromotility in vitro and a 40-60 dB loss of cochlear sensitivity in vivo, without disruption of mechano-electrical transduction in outer hair cells. In heterozygotes, electromotility is halved and there is a twofold (about 6 dB) increase in cochlear thresholds. These results suggest that prestin is indeed the motor protein, that there is a simple and direct coupling between electromotility and cochlear amplification, and that there is no need to invoke additional active processes to explain cochlear sensitivity in the mammalian ear.
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              The aminoglycoside antibiotic dihydrostreptomycin rapidly enters mouse outer hair cells through the mechano-electrical transducer channels.

              The most serious side-effect of the widely used aminoglycoside antibiotics is irreversible intracellular damage to the auditory and vestibular hair cells of the inner ear. The mechanism of entry into the hair cells has not been unequivocally resolved. Here we report that extracellular dihydrostreptomycin not only blocks the mechano-electrical transducer channels of mouse outer hair cells at negative membrane potentials, as previously shown, but also enters the cells through these channels, which are located in the cells' mechanosensory hair bundles. The voltage-dependent blocking kinetics indicate an open-channel block mechanism, which can be well described by a two barrier-one binding site model, quantifying the antibiotic's block of the channel as well as its permeation in terms of the associated rate constants. The results identify the open transducer channels as the main route for aminoglycoside entry. Intracellularly applied dihydrostreptomycin also blocks the transducer channels, but at positive membrane potentials. However, the potency of the block was two orders of magnitude lower than that due to extracellular dihydrostreptomycin. Extracellular Ca2+ increases the free energy of the barrier nearest the extracellular side and of the binding site for dihydrostreptomycin. This reduces both the entry of dihydrostreptomycin into the channel and the channel's affinity for the drug. In vivo, where the extracellular Ca2+ concentration in the endolymph surrounding the hair bundles is < 100 microM, we predict that some 9000 dihydrostreptomycin molecules per second enter each hair cell at therapeutic drug concentrations.
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                Author and article information

                Journal
                Neuron
                Neuron
                Neuron
                Cell Press
                0896-6273
                1097-4199
                23 June 2011
                23 June 2011
                : 70
                : 6
                : 1143-1154
                Affiliations
                [1 ]Department of Biomedical Science, University of Sheffield, Sheffield, S10 2TN, UK
                [2 ]INSERM U587, Université Bordeaux Segalen, CHU Pellegrin, 33076 Bordeaux, France
                [3 ]Department of Neuroscience, University of Wisconsin Medical School, Madison, WI 53706, USA
                Author notes
                []Corresponding author w.marcotti@ 123456sheffield.ac.uk
                [∗∗ ]Corresponding author fettiplace@ 123456wisc.edu
                Article
                NEURON10694
                10.1016/j.neuron.2011.04.024
                3143834
                21689600
                332193bf-ad88-47d6-94af-13b68584ca5e
                © 2011 ELL & Excerpta Medica.

                This document may be redistributed and reused, subject to certain conditions.

                History
                : 26 April 2011
                Categories
                Article

                Neurosciences
                Neurosciences

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