During the last year we have undertaken a detailed study of the signaling pathway that regulates the formation of one specific type of supporting cell, the pillar cell. Pillar cells are only found in mammalian inner ears and the presence of these cells is required for normal auditory function. An examination of expression of members of the fibroblast growth factor signaling pathway indicated that one fgf ligand, Fgf8 is expressed in a limited pattern of cells within the organ of Corti and that one of the fgf receptors, Fgfr3 is expressed in an adjacent population of cells. Interestingly, the cells that express Fgfr3 include cells that will develop as pillar cells. Based on this pattern of expression, we wanted to determine whether signaling between Fgf8 and Fgfr3 might play a role in pillar cell development. As a first step, we inactivated Fgf8 specifically in the ear using a Cre-Lox approach. Analysis of the cochleae from these mice indicated a specific loss of pillar cells. Similarly, when the level of Fgfr3 activation was increased in vitro, the number of cells that developed as pillar cells increased. These results suggest that Fgf8 acts to induce pillar cells through an inductive activation of Fgfr3. Moreover, the fact that increased activation of Fgfr3 leads to an over-production of pillar cells suggests that the level of Fgf8 within the epithelium is normally limiting and that this limitation plays a key role in determining the number and position of pillar cells within the organ of Corti.? ? In a second series of experiments, we analyzed the effects of deletion of the receptor, Fgfr3. As expected, loss of Fgfr3 leads to a similar defect in pillar cell development. This result confirms the inductive interaction between Fgf8 and Fgfr3 in the development of pillar cells. However, in addition to a lack of pillar cells, we observed a number of other defects in Fgfr3 mutant mice. In particular, the Fgfr3-mutant cochleae contain a greater number of hair cells suggesting that some of the cells that would have developed as pillar cells have undergone a fate change to become additional hair cells. A screen for genes with altered expression in Fgfr3 mutants indicated that bone morphogenetic protein 4 (Bmp4) is up-regulated. Since Bmp4 has been shown to influence cell fate, we wanted to determine whether the increase in hair cells might be a result of the increase in Bmp4 signaling. To examine this possibility, Bmp4 signaling within the cochlea was modulated in vitro. Results indicated that increased Bmp4 leads to an increase in hair cells while inhibition of Bmp4 leads to hair cell loss. Moreover, the increased hair cell number in Fgfr3 mutant cochlea can be inhibited if Bmp4 signaling is blocked. These results suggest that a balance between Fgf and Bmp signaling may play a role in regulating the number of pillar cells versus hair cells within the developing organ of Corti.
|Puligilla, Chandrakala; Feng, Feng; Ishikawa, Kotaro et al. (2007) Disruption of fibroblast growth factor receptor 3 signaling results in defects in cellular differentiation, neuronal patterning, and hearing impairment. Dev Dyn 236:1905-17|
|Jacques, Bonnie E; Montcouquiol, Mireille E; Layman, Erynn M et al. (2007) Fgf8 induces pillar cell fate and regulates cellular patterning in the mammalian cochlea. Development 134:3021-9|