The Stereociliary Paracrystal Is a Dynamic Cytoskeletal Scaffold In Vivo

• Transgenic zebrafish reveal stereociliary protein dynamics in vivo
• Actin cross-linker fascin 2b exchange within the stereocilium is rapid and unbiased
• Stereocilia exchange β-actin and myosin XVa in the shafts and tips, respectively
• Protein dynamism is suggested to underpin stereociliary function and longevity

SummaryPermanency of mechanosensory stereocilia may be the consequence of low protein turnover or rapid protein renewal. Here, we devise a system, using optical techniques in live zebrafish, to distinguish between these mechanisms. We demonstrate that the stereocilium’s abundant actin cross-linker fascin 2b exchanges, without bias or a phosphointermediate, orders of magnitude faster (t1/2 of 76.3 s) than any other known hair bundle protein. To establish the logic of fascin 2b’s exchange, we examine whether filamentous actin is dynamic and detect substantial β-actin exchange within the stereocilium’s paracrystal (t1/2 of 4.08 hr). We propose that fascin 2b’s behavior may enable cross-linking at fast timescales of stereocilia vibration while noninstructively facilitating the slower process of actin exchange. Furthermore, tip protein myosin XVa fully exchanges in hours (t1/2 of 11.6 hr), indicating that delivery of myosin-associated cargo occurs in mature stereocilia. These findings suggest that stereocilia permanency is underpinned by vibrant protein exchange.

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