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Volume 16, Number 17, Issue of September 1, 1996 pp. 5466-5477
Copyright ©1996 Society for Neuroscience

Regenerative Proliferation in Organ Cultures of the Avian Cochlea: Identification of the Initial Progenitors and Determination of the Latency of the Proliferative Response

Received Jan. 18, 1996; revised May 31, 1996; accepted June 5, 1996.

Mark E. Warchol¹ and Jeffrey T. Corwin^{1, 2}

¹ Departments of Otolaryngology-Head and Neck Surgery and ² Neuroscience, School of Medicine, University of Virginia, Charlottesville, Virginia 22908

Sensory hair cells in the cochleae of birds are regenerated after the death of preexisting hair cells caused by acoustic overstimulation or administration of ototoxic drugs. Regeneration involves renewed proliferation of cells in an epithelium that is otherwise mitotically quiescent. To determine the identity of the first cells that proliferate in response to the death of hair cells and to measure the latency of this proliferative response, we have studied hair-cell regeneration in organ culture. Cochleae from hatchling chicks were placed in culture, and hair cells were killed individually by a laser microbeam. The culture medium was then replaced with a medium that contained a labeled DNA precursor. The treated cochleae were incubated in the labeling media for different time periods before being fixed and processed for the visualization of proliferating cells. The first cells to initiate DNA replication in response to the death of hair cells were supporting cells within the cochlear sensory epithelium. All of the labeled supporting cells were located within 200 µm of the hair-cell lesions. These cells first entered S-phase ~16 hr after the death of hair cells. The results indicate that supporting cells are the precursors of regenerated hair cells and suggest that regenerative proliferation of supporting cells is triggered by signals that act locally within the damaged epithelium.

Key words: hair cell; hearing; ear; regeneration; cochlea; auditory system; precursor

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