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Among the vertebrate lineages with different hearing frequency ranges, humans lie between the low-frequency hearing (1 kHz) of fish and amphibians and the high-frequency hearing (100 kHz) of bats and dolphins. Little is known about the mechanism underlying such a striking difference in the limits of hearing frequency. Prestin, responsible for cochlear amplification and frequency selectivity in mammals, seems to be the only candidate to date. Mammalian prestin is densely expressed in the lateral plasma membrane of the outer hair cells (OHCs) and functions as a voltage-dependent motor protein. To explore the molecular basis for the contribution of prestin in hearing frequency detection, we collected audiogram data from humans, dogs, gerbils, bats, and dolphins because their average hearing frequency rises in steps. We generated stable cell lines transfected with human, dog, gerbil, bat, and dolphin prestins (hPres, dPres, gPres, bPres, and nPres, respectively). The non-linear capacitance (NLC) of different prestins was measured using a whole-cell patch clamp. We found that the  Q  max / C  lin  of bPres and nPres was significantly higher than that of humans. The  V  1  /  2  of hPres was more hyperpolarized than that of nPres. The  z  values of hPres and bPres were higher than that of nPres. We further analyzed the relationship between the high-frequency hearing limit ( F  max ) and the functional parameters ( V  1  /  2 ,  z , and  Q  max / C  lin ) of NLC among five prestins. Interestingly, no significant correlation was found between the functional parameters and  F  max . Additionally, a comparative study showed that the amino acid sequences and tertiary structures of five prestins were quite similar. There might be a common fundamental mechanism driving the function of prestins. These findings call for a reconsideration of the leading role of prestin in hearing frequency perception. Other intriguing kinetics underlying the hearing frequency response of auditory organs might exist.

Functional Parameters of Prestin Are Not Correlated With the Best Hearing Frequency