The mammalian inner ear contains two sensory organs, the cochlea and vestibule. Their sensory neuroepithelia are characterized by a mosaic of hair cells and supporting cells. Cochlear hair cells differentiate in four rows: a single row of inner hair cells (IHCs) and three rows of outer hair cells (OHCs). Recent studies have shown that Math1, a mammalian homolog of Drosophila atonal is a positive regulator of hair cell differentiation. The basic helix–loop–helix (bHLH) genes Hes1 andHes5 (mammalian hairy andEnhancer-of-split homologs) can influence cell fate determination by acting as negative regulators to inhibit the action of bHLH-positive regulators. We show by using reverse transcription-PCR analysis that Hes1, Hes5, andMath1 are expressed in the developing mouse cochleae.In situ hybridization revealed a widespread expression of Hes1 in the greater epithelial ridge (GER) and in lesser epithelial ridge (LER) regions. Hes5 is predominantly expressed in the LER, in supporting cells, and in a narrow band of cells within the GER.

Examination of cochleae fromHes1^−/− mice showed a significant increase in the number of IHCs, whereas cochleae fromHes5^−/− mice showed a significant increase in the number of OHCs. In the vestibular system, targeted deletion of Hes1 and to a lesser extentHes5 lead to formation of supernumerary hair cells in the saccule and utricle.

The supernumerary hair cells in the mutant mice showed an upregulation of Math1. These data indicate that Hes1 andHes5 participate together for the control of inner ear hair cell production, likely through the negative regulation of Math1.