ABSTRACTEarly postnatal mouse cochlear cultures were treated with a small panel of kinase inhibitors to elucidate the mechanisms underlying the maintenance of hair‐bundle structure in the developing inner ear. At low concentrations (1–10 nM), staurosporine causes the collapse and loss of hair bundles without provoking hair‐cell death, as judged by lack of terminal transferase dUTP nick end labeling (TUNEL) labeling or reactivity to anti‐activated caspase‐3. Staurosporine exposure results in the fusion of the hair bundle's stereocilia, a resorption of the parallel actin bundles of the stereocilia into the cytoplasm of the hair cell, a detachment of the apical, non‐stereociliary membrane of the hair cell from the underlying cuticular plate, and a severing of the hair‐bundle's rootlets from the actin cores of the stereocilia. It does not block membrane retrieval at the apical pole of the hair cells, nor does it elicit the externalization of phosphatidylserine. Staurosporine treatment causes a reduction in levels of the phosphorylated forms of ezrin, radixin, and moesin in cochlear cultures during the period of hair‐bundle loss, indicating the integrity of the hair bundle may be actively maintained by the phosphorylation status of these proteins. J. Comp. Neurol. 522:3281–3294, 2014. © 2014 The Authors The Journal of Comparative Neurology Published by Wiley Periodicals, Inc.