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Role of Jagged1 in cochlear supporting cell differentiation and maintenance

ABSTRACT: Inner ear cochlear supporting cells (SCs) are highly specialized glia-like cells that structurally and functionally support neighboring mechano-sensory hair cells. Despite their importance for proper auditory function, little is known about the molecular mechanisms that control their development. In this study we investigated the function of the Notch ligand Jagged1 (Jag1) in cochlear SC differentiation and maintenance. To address the function of Jag1 in the differentiation of SCs, we conditionally deleted Jag1 in stage E14.5 SC precursors using the recently established Sox2-CreER/+ and Jag1 fx/fx mouse lines. Analysis of stage P0 Jag1 mutant and control animals revealed that Hensen cells, a highly specialized SC-subtype located at the lateral edge of the auditory sensory epithelium, failed to form in the absence of Jag1. Other SC-subtypes did form in the absence of Jag1, however, their morphology and cellular arrangement was abnormal and SC-subtype specific genes and genes associated with mitochondrial function and protein synthesis were significantly reduced, indicating global defects in SC differentiation and SC homeostasis.

ORGANISM(S): Mus musculus

PROVIDER: GSE148009 | GEO | 2020/12/16

REPOSITORIES: GEO

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Project description:We have employed a novel approach for the identification of functionally important microRNA (miRNA)-target interactions using integrated miRNA, transcriptome and proteome profiles with advanced in silico analysis. By looking at both the transcript and protein levels of expression, a thorough coverage of miRNA regulation was obtained. Microdissected auditory and vestibular sensory epithelia were used as the model system, thus being the first time such a comparison was carried out in a neuroepithelial system. Moreover, this is one of only a few studies employing proteome screening for the identification of miRNA targets. Notably, this approach can be employed for the study of other tissues and organs. We detected the expression of 157 miRNAs in the inner ear sensory epithelia, with 53 miRNAs differentially expressed between the cochlea and vestibule. By searching for enrichment and depletion of miRNA targets in the transcript and protein datasets with a reciprocal or similar expression, respectively, as the regulatory miRNA, we identified functionally important miRNAs. Finally, the interaction between miR-135b and PSIP1-P75, a transcriptional coacitvator previously unknown in the inner ear, was identified and validated experimentally. We suggest that miR-135b may serve as a cellular effector, involved in regulating some of the differences between the cochlear and vestibular hair cells. We investigated the mRNA expression profile of the cochlear and vestibular sensory epithelia from inner ears of postnatal day 2 mice using the Affymetrix GeneChip® 430 2Mouse Genome array. Cochlear and vestibular sensory epithelia were dissected from wild type C3H mice and collected separately. The vestibular epithelia consisted of the saccule, utricle and the lateral and anterior cristae. Both the cochlear and vestibular sensory epithelia were dissected with their underlying mesenchyma. Altogether three pools, three biological replicates, of each tissue type were collected consisting of cochlear or vestibular sensory epithelia dissected from 10 to 12 inner ears.

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Role of Jagged1 in cochlear supporting cell differentiation and maintenance

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