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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2019, Vol. 13 Issue (6) : 705-712    https://doi.org/10.1007/s11684-019-0703-y
RESEARCH ARTICLE
Unidirectional and stage-dependent roles of Notch1 in Wnt-responsive Lgr5+ cells during mouse inner ear development
Hui Jiang1,3, Shan Zeng1,2, Wenli Ni1,2, Yan Chen1,2(), Wenyan Li1,2()
1. ENT Institute and Otorhinolaryngology Department of Affiliated Eye and ENT Hospital, Fudan University, Shanghai 200031, China
2. Key Laboratory of Hearing Medicine of NHFPC, Shanghai 200031, China
3. Otorhinolaryngology Department of Jinshan Hospital, Fudan University, Shanghai 201508, China
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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     
Corresponding Authors: Yan Chen,Wenyan Li   
Just Accepted Date: 08 August 2019   Online First Date: 10 October 2019    Issue Date: 16 December 2019
 Cite this article:   
Hui Jiang,Shan Zeng,Wenli Ni, et al. Unidirectional and stage-dependent roles of Notch1 in Wnt-responsive Lgr5+ cells during mouse inner ear development[J]. Front. Med., 2019, 13(6): 705-712.
 URL:  
http://academic.hep.com.cn/fmd/EN/10.1007/s11684-019-0703-y
http://academic.hep.com.cn/fmd/EN/Y2019/V13/I6/705
Fig.1  EGFP immunofluorescence in Lgr5-EGFP-creER mice showed that Lgr5 was expressed in the cochlear duct at E13.5. Sox2 and Jag1 were used as markers of cochlear prosensory epithelium. The images of the lower magnification (A) and the higher magnification of apical turn (B) are shown. Scale bar: (A) 100 µm; (B) 50 µm.
Fig.2  Notch inhibition at E10.5 in Lgr5+ cells showed no evident effect on the expression pattern of P27kip1. (A–D) Typical immunofluorescence images of control cochleae at E14.5. (E–H) Typical immunofluorescence images of Lgr5 CreERT2/+;Notch1flox/flox cochleae at E14.5. Tamoxifen was administered at E10.5, and specimens were harvested at E14.5. Scale bars: (A, D) 80 µm; (B–D, E–H) 40 µm.
Fig.3  Notch1 inhibition at E14.5 drove the Lgr5+ cells in the pillar cell region to re-enter the cell cycle and differentiate into new HCs. Tamoxifen was administered at E14.5, and specimens were harvested at E18.5. Typical XYZ (A, C, E, and G) and XZY (B, D, F, and H) images of the apical and middle turn in the cochlear epithelium from the control and Lgr5 CreERT2/+;Notch1flox/flox mice are shown. The typical images of whole cochleae in each group are shown in I and J. The white box indicated the position of A in I and E in J. The arrows indicated the extra new hair cells (HCs). HC layer: HC layer. SC layer: supporting cell layer. Scale bars: (A–H) 20 µm; (I, J) 300 µm.
Fig.4  Notch1 overexpression in embryonic Lgr5+ cells showed no evident effect on cochlear sensory epithelium formation or cell differentiation. (A, B) Typical immunofluorescence images of the apical turn in the cochlear epithelium from control and Lgr5 CreERT2/+;Notch1OE/+ cochleae at E14.5. Tamoxifen was administered at E10.5, and specimens were harvested at E14.5. (C, D) Typical immunofluorescence images of the apical and middle turn in the cochlear epithelium from control cochleae and Lgr5 CreERT2/+;Notch1OE/+ cochleae at E18.5. Tamoxifen was administered at E14.5, and specimens were harvested at E18.5. Scale bars: (A, B) 50 µm; (C, D) 20 µm.
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