Journal
NATURE COMMUNICATIONS
Volume 6, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/ncomms7873
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Funding
- National Institutes of Health/National Institute of Biomedical Imaging and Bioengineering (NIH/NIBIB) [P41RR14579, P41EB001974]
- NIH [R01DC00033, R01DC03463, R01DC04179, R37DK39773, R01EY12963, R01GM47214, R01D K58762]
- National Science Foundation Division of Integrative Biology and Neuroscience (NSF/IBN) [IBN-998298]
- National Institutes of Health intramural funds from the National Institute on Deafness and Other Communication Disorders [DC000079-02, DC000039-17]
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The maintenance of sensory hair cell stereocilia is critical for lifelong hearing; however, mechanisms of structural homeostasis remain poorly understood. Conflicting models propose that stereocilia F-actin cores are either continually renewed every 24-48 h via a treadmill or are stable, exceptionally long-lived structures. Here to distinguish between these models, we perform an unbiased survey of stereocilia actin dynamics in more than 500 utricle hair cells. Live-imaging EGFP-beta-actin or dendra2-beta-actin reveal stable F-actin cores with turnover and elongation restricted to stereocilia tips. Fixed-cell microscopy of wild-type and mutant beta-actin demonstrates that incorporation of actin monomers into filaments is required for localization to stereocilia tips. Multi-isotope imaging mass spectrometry and live imaging of single differentiating hair cells capture stereociliogenesis and explain uniform incorporation of N-15-labelled protein and EGFP-beta-actin into nascent stereocilia. Collectively, our analyses support a model in which stereocilia actin cores are stable structures that incorporate new F-actin only at the distal tips.
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