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Live-cell imaging of actin dynamics reveals mechanisms of stereocilia length regulation in the inner ear

Author

Listed:
  • Meghan C. Drummond

    (Laboratory of Molecular Genetics, Section on Human Genetics, National Institute on Deafness and Other Communication Disorders, National Institutes of Health)

  • Melanie Barzik

    (Laboratory of Molecular Genetics, Section on Human Genetics, National Institute on Deafness and Other Communication Disorders, National Institutes of Health)

  • Jonathan E. Bird

    (Laboratory of Molecular Genetics, Section on Human Genetics, National Institute on Deafness and Other Communication Disorders, National Institutes of Health)

  • Duan-Sun Zhang

    (Harvard Medical School and Howard Hughes Medical Institute)

  • Claude P. Lechene

    (National Resource for Imaging Mass Spectrometry, Brigham and Women’s Hospital and Harvard Medical School
    Brigham and Women’s Hospital and Harvard Medical School)

  • David P. Corey

    (Harvard Medical School and Howard Hughes Medical Institute)

  • Lisa L. Cunningham

    (Section on Sensory Cell Biology, National Institute on Deafness and Other Communication Disorders, National Institutes of Health)

  • Thomas B. Friedman

    (Laboratory of Molecular Genetics, Section on Human Genetics, National Institute on Deafness and Other Communication Disorders, National Institutes of Health)

Abstract

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-β-actin or dendra2-β-actin reveal stable F-actin cores with turnover and elongation restricted to stereocilia tips. Fixed-cell microscopy of wild-type and mutant β-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 15N-labelled protein and EGFP-β-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.

Suggested Citation

  • Meghan C. Drummond & Melanie Barzik & Jonathan E. Bird & Duan-Sun Zhang & Claude P. Lechene & David P. Corey & Lisa L. Cunningham & Thomas B. Friedman, 2015. "Live-cell imaging of actin dynamics reveals mechanisms of stereocilia length regulation in the inner ear," Nature Communications, Nature, vol. 6(1), pages 1-10, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms7873
    DOI: 10.1038/ncomms7873
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