The middle ear epithelium is derived from neural crest and endoderm, which line distinct regions of the middle ear cavity. Here we investigate the distribution of putative stem cell markers in the middle ear combined with an analysis of the location of label retaining cells (LRCs) to create a map of the middle ear mucosa. We show that proliferating cells and LRCs were associated with specific regions of the ear epithelium, concentrated in the hypotympanum at the base of the auditory bulla and around the ear drum. Sox2 was widely expressed in the endodermally-derived ciliated pseudostratified epithelium of the hypotympanum. This part of the middle ear showed high levels of Wnt activity, as indicated by the expression of Axin 2, a readout of Wnt signaling. Keratin 5 showed a more restricted expression within the basal cells of this region, with very little overlap between the Sox2 and Keratin 5 positive epithelium, indicating that these genes mark distinct populations. Little expression of Sox2 or Keratin 5 was observed in the neural crest-derived middle ear epithelium that lined the promontory, except in cases of otitis media when this epithelium underwent hyperplasia. This study lays the foundation for further understanding of homeostasis and repair in the middle ear.
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