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Related Experiment Video

Updated: Nov 5, 2025

In Ovo and Ex Ovo Methods to Study Avian Inner Ear Development

In Ovo and Ex Ovo Methods to Study Avian Inner Ear Development

Published on: June 16, 2022

Cochlea cell-specific marker expression upon in vitro Hes1 knockdown.

A C Batissoco^1,2, K Lezirovitz^1,2, D B Zanatta³

¹Otorrinolaringologia/LIM32, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brasil.

Brazilian Journal of Medical and Biological Research = Revista Brasileira De Pesquisas Medicas E Biologicas

|May 19, 2021

Summary

This summary is machine-generated.

Knocking down Hes1 in the inner ear organ of Corti cultures increased hair cell and progenitor cell markers. This suggests Hes1 inhibition is a promising strategy for hair cell regeneration.

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Area of Science:

Developmental biology
Oto-science
Molecular biology

Background:

The NOTCH pathway, involving HES1, regulates inner ear development via lateral inhibition.
This mechanism maintains supporting cell phenotypes and prevents them from becoming hair cells.
Genetic manipulation of the NOTCH pathway can increase hair cell numbers.

Purpose of the Study:

To investigate the gene expression changes in hair cells and supporting cells following Hes1 knockdown.
To evaluate the efficacy of Hes1-shRNA lentivirus in organotypic cultures of the organ of Corti.

Main Methods:

Organotypic cultures of the organ of Corti from postnatal-day-3 mice were used.
Hes1 knockdown was achieved using Hes1-shRNA lentivirus transduction.
Gene expression levels (mRNA and protein) were analyzed 48 hours post-transduction.

Main Results:

Hes1 gene expression was significantly reduced at both mRNA and protein levels post-knockdown.
mRNA levels of Myo7a (hair cell marker) and Sox2 (progenitor cell marker) significantly increased.
Observed gene expression modulation aligns with interference in the lateral inhibition mechanism.

Conclusions:

Hes1 knockdown effectively modulates gene expression in the organ of Corti.
Lentivirus-mediated Hes1-shRNA expression is a viable strategy for genetic interference in the organ of Corti.
This approach holds potential for future in vivo hair cell regeneration strategies.