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The authors use post-genomic experimentation that employs physiological measurements in mouse mutants to elucidate a role for myosin 7a in mechanotransduction in auditory hair cells. For the MET channels to open, the hair bundles in the mutants must be deflected several microns beyond their normal operating range, which suggests that myosin 7a normally positions the bundle for optimal transducer sensitivity.
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This study uses both yeast two-hybrid and pull-down binding assays to demonstrate interactions among three proteins (myosin 7a, cadherin-23 and a PDZ protein harmonin) that are known to be mutated in Usher type I deaf-blindness syndrome. Equivalent mutations in mice cause defective hair bundle development and result in deafness. The authors conclude that myosin 7a ferries harmonin along the actin core of developing stereocilia, and that the three-protein complex is essential for stereociliary cohesion.
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Boëda B., El-Amraoui A., Bahloul A., Goodyear R., Daviet L., Blanchard S., Perfettini I., Fath K.R., Shorte S., Reiners J.et al. Myosin VIIa, harmonin and cadherin 23, three Usher I gene products that cooperate to shape the sensory hair bundle. EMBO J. 21:2002;6689-6699 This study uses both yeast two-hybrid and pull-down binding assays to demonstrate interactions among three proteins (myosin 7a, cadherin-23 and a PDZ protein harmonin) that are known to be mutated in Usher type I deaf-blindness syndrome. Equivalent mutations in mice cause defective hair bundle development and result in deafness. The authors conclude that myosin 7a ferries harmonin along the actin core of developing stereocilia, and that the three-protein complex is essential for stereociliary cohesion.
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The authors present the most detailed account so far of the properties and mechanisms of spontaneous hair bundle oscillations. They demonstrate that they are driven by the slow adaptation motor biasing the displacement-force relation of the hair bundle into a region of negative slope.
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The authors describe a single-cell correlate of the electrically evoked otoacoustic emissions that are recorded in most vertebrate classes and used as evidence for ubiquitous active mechanical output from the hair cells. Spontaneous oscillations of saccular hair bundles were modulated at frequencies of up to 1 kHz by extracellular electrical stimulation. The specificity of the response was tied to hair cell transduction by showing that it disappeared on blocking the MET channels.
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The authors demonstrate that MET currents in turtle auditory hair cells tuned to high and low frequencies show different sensitivities to channel blockers such as dihydrostreptomycin, and have different open times as assessed by the spectra of the current noise.
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The authors found that mutagenesis of many of the amino acids from charged to neutral in the putative membrane domain of the prestin molecule had little or no effect on the voltage sensitivity of the membrane protein. This led them to conclude that the voltage sensor for the outer hair cell motor was a charged particle extrinsic to the protein, and they identified it as arising from the binding of intracellular chloride and bicarbonate ions.
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Oliver D., He D.Z.Z., Klöcker N., Ludwig J., Schulte U., Waldegger S., Ruppersberg J.P., Dallos P., Fakler B. Intracellular anions as the voltage sensor or prestin, the outer hair cell motor protein. Science. 292:2001;2340-2343 The authors found that mutagenesis of many of the amino acids from charged to neutral in the putative membrane domain of the prestin molecule had little or no effect on the voltage sensitivity of the membrane protein. This led them to conclude that the voltage sensor for the outer hair cell motor was a charged particle extrinsic to the protein, and they identified it as arising from the binding of intracellular chloride and bicarbonate ions.
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The authors found that targeted deletion of prestin in mice abolishes the somatic motility of outer hair cells in vitro and causes a 40-60 dB loss of auditory sensitivity and otoacoustic emissions in vivo without affecting the MET channels. This is strong evidence for prestin being part of the electromechanical feedback loop that underlies cochlear amplifier.
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