Unidirectional and stage-dependent roles of Notch1 in Wnt-responsive Lgr5⁺ cells during mouse inner ear development

Abstract

Wnt and Notch signaling play crucial roles in the determination of the prosensory domain and in the differentiation of hair cells (HCs) and supporting cells during mouse inner ear development; however, the relationship between the two signaling pathways in the mouse cochlea remains largely unknown. Here, we investigated the interactions between Notch and Wnt signaling on the basis of the bidirectional regulation of Notch1 specifically in Wnt-responsive Lgr5⁺ progenitors during different cochlear development stages. We found that the downregulation of Notch1 in Lgr5⁺ cells from embryonic day (E) 14.5 to E18.5 can drive the quiescent Lgr5⁺ cells to re-enter the cell cycle and differentiate into extra HCs, whereas the upregulation of Notch1 expression did not affect the proliferation or differentiation of otic progenitor cells. No effect was observed on the upregulation or downregulation of Notch1 in Lgr5⁺ cells from E10.5 to E14.5. We concluded that the roles of Notch1 in Wnt-responsive Lgr5⁺ cells are unidirectional and stage dependent and Notch1 serves as a negative regulator for Lgr5⁺ progenitor activation during cochlear differentiation. Our findings improved the understanding of the interactions between Notch and Wnt signaling in cochlear development.

Keywords

inner ear / cochlear / Wnt / Notch / Lgr5 / auditory system

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Acknowledgements

The authors wish to thank Wen Li for providing technical support. This work was supported by the National Key R&D Program of China (No. 2017YFA0103900), the National Natural Science Foundation of China (Nos. 81771011, 81771010, 81700910, 81700914, and 81400463), the Development Fund for Shanghai Talents (No. 2017046), the Excellent Personnel Training Plan for Shanghai Health System (No. 2017Q003), and the Shanghai HFPC Foundation (No. 201440402).

Compliance with ethics guidelines

Hui Jiang, Shan Zeng, Wenli Ni, Yan Chen, and Wenyan Li declare that they have no conflict of interest. All institutional and national guidelines for the care and use of laboratory animals were followed. This study was carried out in strict accordance with the Guiding Directive for Humane Treatment of Laboratory Animals’ issued by the Chinese National Ministry of Science and Technology on September, 2006. All experiments were approved by the Shanghai Medical Experimental Animal Administrative Committee (Permit Number: 2009-0082). All efforts were made to minimize suffering and reduce the number of animals used.

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