Title: The sensitivity of the cochlear amplifier to changes in operating conditions
Description: Frequency selectivity is one of the most important functions of the mammalian hearing organ – the cochlea. The interaction of fluid mass and organ of Corti compliance sets a traveling wave along the basilar membrane (BM), which is longitudinally tuned to different frequencies. Beyond this passive tuning process, cochlear amplification locally enhances the vibration of the best frequency peak by factors of hundreds to boost the frequency selectivity and sensitivity of the cochlea. This amplification is achieved by a positive feedback loop between BM motion and outer hair cell (OHC) electrical-mechanical response. However, this active mechanism is vulnerable to damage and cannot be fully recovered in vivo. As the instruments of cochlear amplification, the frequency response of BM and OHCs are of great importance to understand cochlear tuning process. This thesis used animal models, aimed to understand cochlear tuning and investigate possibilities to manipulate the cochlear amplifier, by testing the cochlear amplifier’s sensitivity to operating conditions.

The first project tested whether the cochlear amplification can adjust to a lower endocochlear potential (EP), which controls OHC electromechanical force by providing part of the voltage source to drive OHC transduction current. To investigate this possibility, we use intraperitoneal (IP) and intravenous (IV) injection of furosemide to reversibly reduce EP, while monitoring the EP and cochlear amplification simultaneously. Cochlear amplification was monitored by measuring the local cochlear microphonic (LCM) and distortion product emission (DPOAE). With IV injection, the cochlear amplification observed in LCM could attain nearly full or even full recovery with reduced EP. This showed the cochlea has an ability to adjust to diminished operating condition. Furthermore, the cochlear amplifier and EP recovered with different time courses: cochlear amplification just started to recover after the EP was nearly fully recovered and stabilized. Using a Boltzmann model and the 2nd harmonic of the LCM to estimate the mechanoelectric transducer channel operating point, we found that the recovery of cochlear amplification occurred with re-centering of the operating point.

The second project studied the physiological and anatomical effects of perfusing the cochlea with a viscous fluid, for better understanding cochlear fluid mechanics. Perilymphatic perfusion was applied with artificial perilymph and viscous sodium hyaluronate (Healon, HA) in four different concentrations. Using compound action potential (CAP) thresholds as an indicator of cochlear condition, our results and analysis indicated that the cochlea can sustain, without a significant CAP threshold shift, up to a 1.5 Pa shear stress. Histology of the cochleae perfused with higher shear stress showed the Reissner's membrane was torn. These data also indicated that the cochlea mechanics remains normal within increased perilymphatic fluid viscosity up to an increase of a factor of 50. Beside these findings, a temporary CAP threshold shift was observed, perhaps due to the presence and then clearance of viscous fluid within the cochlea, or to a temporary position shift of the organ of Corti. The last project was to test the effect of OHC intracellular Cl- concentration on cochlear amplification.

Chloride is known to enable the electromotility of the OHC by binding its motor protein, prestin. By locally perfusing high chloride perilymph and the chloride ionophore tributyltin, this study investigated whether increasing intracellular chloride concentration can restore cochlear sensitivity in a cochlea that was slightly damaged. This had been shown by others in guinea pig. However, we did not observe recovery in several attempts in gerbil.