Involvement of Dmp1 in the Precise Regulation of Hair Bundle Formation in the Developing Cochlea

SIMPLE SUMMARY: Deafness is a common clinical disease and a major global public health problem worldwide. The hair cell regeneration is one of the most promising strategies to address hearing loss. Thus, it is urgent to explore the development and differentiation process of hair cells at the cellular and molecular level, which is the basis of “overall regeneration of functional auditory receptors”. We observed the characteristics of Dmp1 involved in inner ear morphogenesis by using recombinant enzyme (Cre) transgenic mice to knockdown Dmp1 in early inner ear development. In our study, we found Dmp1 was already expressed early in inner ear development and Dmp1 mutant mice had abnormal hair cell morphology (disordered stereocilia and mislocalized kinococilia). It is reasonable to think that Dmp1 plays a role in the precise regulation of hair bundle morphogenesis. Dmp1 may be a suitable target molecule for the treatment of deafness, and our study provides strong support for gene therapy for sensorineural deafness. ABSTRACT: Dentin matrix protein 1 (Dmp1) is a highly phosphorylated, extracellular matrix protein that is extensively expressed in bone and teeth but also found in soft tissues, including brain and muscle. However, the functions of Dmp1 in the mice cochlea are unknown. Our study showed that Dmp1 was expressed in auditory hair cells (HCs), with the role of Dmp1 in those cells identified using Dmp1 cKD mice. Immunostaining and scanning electron microscopy of the cochlea at P1 revealed that Dmp1 deficiency in mice resulted in an abnormal stereociliary bundle morphology and the mispositioning of the kinocilium. The following experiments further demonstrated that the cell-intrinsic polarity of HCs was affected without apparent effect on the tissue planer polarity, based on the observation that the asymmetric distribution of Vangl2 was unchanged whereas the Gαi3 expression domain was enlarged and Par6b expression was slightly altered. Then, the possible molecular mechanisms of Dmp1 involvement in inner ear development were explored via RNA-seq analysis. The study suggested that the Fgf23–Klotho endocrine axis may play a novel role in the inner ear and Dmp1 may regulate the kinocilium–stereocilia interaction via Fgf23–Klotho signaling. Together, our results proved the critical role of Dmp1 in the precise regulation of hair bundle morphogenesis in the early development of HCs.