Actin at stereocilia tips is regulated by mechanotransduction and ADF/cofilin

Stereocilia on auditory sensory cells are actin-based protrusions that mechanotransduce sound into an electrical signal. These stereocilia are arranged into a bundle with three rows of increasing length to form a staircase-like morphology that is required for hearing. Stereocilia in the shorter rows, but not the tallest row, are mechanotransducing because they have force-sensitive channels localized at their tips. The onset of mechanotransduction during mouse postnatal development refines stereocilia length and width. However, it is unclear how actin is differentially regulated between stereocilia in the tallest row of the bundle and the shorter, mechanotransducing rows. Here, we show actin turnover is increased at the tips of mechanotransducing stereocilia during bundle maturation. Correspondingly, from birth to postnatal day 6, these stereocilia had increasing amounts of available actin barbed ends, where monomers can be added or lost readily, as compared with the non-mechanotransducing stereocilia in the tallest row. The increase in available barbed ends depended on both mechanotransduction and MYO15 or EPS8, which are required for the normal specification and elongation of the tallest row of stereocilia. We also found that loss of the F-actin-severing proteins ADF and cofilin-1 decreased barbed end availability at stereocilia tips. These proteins enriched at mechanotransducing stereocilia tips, and their localization was perturbed by the loss of mechanotransduction, MYO15, or EPS8. Finally, stereocilia lengths and widths were dysregulated in Adf and Cfl1 mutants. Together, these data show that actin is remodeled, likely by a severing mechanism, in response to mechanotransduction. •Mechanotransduction regulates actin at the tips of mammalian cochlear stereocilia•With transduction onset, ADF/cofilin localize to mechanotransducing stereocilia tips•Actin-severing proteins increase available F-actin barbed ends at stereocilia tips•Normal stereocilia length and width depend on ADF and cofilin-1 McGrath et al. show that ADF/cofilin-1 are recruited to mechanotransducing auditory stereocilia tips, where they increase the availability of F-actin barbed ends. Blocking transduction either by constitutive mutation of the channel or acute inhibition in cochlear explants perturbed ADF/cofilin-1 localization and reduced barbed end availability.