Katie Peterson Phy Sci 173: Section 1A 12 April 2010 Prestin-Mediated Electromotility of Mammalian Outer Hair Cells Along

with mechano-electrical transduction, the mammalian outer hair cell (OHC) performs electromechanical transduction, which could be responsible for cochlear amplification. Prestin is hypothesized to be the molecular motor driving the OHC electromotility, and thus allowing the mechanical sensitivity of the mammalian cochlea. This study was a quantitative analysis of hair cell and cochlear function in prestin mutant mice. They created a mutant mouse by targeted disruption of the prestin gene. Genomic Southern analysis and PCR analysis confirmed the targeted deletion. RT-PCR analysis of deleted region showed that heterozygote cochlea had half the amount of prestin messenger RNA found in normal cochlea, while no transcript was found in the cochlea of prestin null mice. Mutant mice displayed shorter OHC lengths and a loss of inner and outer hair cells in basal quarter of cochlea, while heterozygous mice displayed intermediate OHC lengths and had scattered loss of hair cells in the same region. To determine if prestin is crucial for normal electromotility, OHCs were isolated and tested for motility through the use of a voltage clamp. Normal OHCs show length changes when the membrane voltage is altered, but OHCs from mutant mice did not show motility at any point, while heterozygous OHC absolute motility was about half of the wild type. Hair bundles, which allow for mechano-electrical transduction, were normal in prestin null mice. Also, cochlear microphonic (CM), which provides a measure of transducer function in OHCs, is an electrical response dominated by OHC receptor potential. The observed CM amplitudes suggest mechano electrical transduction in mutant OHCs still occurs, implying the decrease in cochlear sensitivity is due to the loss of electromotility in the mutant OHCs. In vivo hair cell response showed that OHC electromotility was necessary for normal cochlear amplification. In the absence of prestin, there is a loss of electromotility in vitro as well as a 40-60 dB decrease in cochlear sensitivity in vivo. Heterozygous cells displayed half the electromotility of normal cells, along with a twofold increase in cochlear thresholds. Thus, prestin is required for both OHC electromotility in vitro and normal cochlear amplification in vivo. These results suggest that OHC electromotility is the force generator for the cochlear amplifier. There may be other contributors to cochlear amplification, such as hair bundles, which may contribute to mechanical amplification in lower vertebrates, as non-mammalian hair cells lack electromotility. The article concludes that prestin dependent electromotility of OHC singlehandedly provides the cochlear amplification needed to explain the cochlear sensitivity of mammals, suggesting no other active processes are needed in the cochlear amplification mechanism.