Cell Type–Specific Transcriptome Analysis Reveals a Major Role for Zeb1 and miR-200b in Mouse Inner Ear Morphogenesis
| Autor: | Scott E. Strome, Kiyoto Kurima, Andrew J. Griffith, Michelle A. Sallin, Ran Elkon, Siaw-Lin Chan, Douglas S. Darling, David J. Eisenman, Andrew Biedlingmaier, Annie Morrisson, Ronna Hertzano |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Cancer Research
Gene Expression QH426-470 Deafness Transcriptome Mesoderm Mice 0302 clinical medicine Morphogenesis Genetics (clinical) Inner ear morphogenesis Vestibular system Regulation of gene expression Genetics 0303 health sciences Mice Inbred ICR Systems Biology Gene Expression Regulation Developmental Genomics Sensory Systems Cell biology medicine.anatomical_structure Auditory System Vestibule Labyrinth Research Article Kruppel-Like Transcription Factors Biology Molecular Genetics 03 medical and health sciences medicine otorhinolaryngologic diseases Animals Inner ear Gene Networks Molecular Biology Ecology Evolution Behavior and Systematics 030304 developmental biology Homeodomain Proteins Gene Expression Profiling Zinc Finger E-box-Binding Homeobox 1 Epithelial Cells Gene expression profiling MicroRNAs Ear Inner Genetics of Disease sense organs Genome Expression Analysis 030217 neurology & neurosurgery Neuroscience |
| Zdroj: | PLoS Genetics PLoS Genetics, Vol 7, Iss 9, p e1002309 (2011) |
| ISSN: | 1553-7404 1553-7390 |
| Popis: | Cellular heterogeneity hinders the extraction of functionally significant results and inference of regulatory networks from wide-scale expression profiles of complex mammalian organs. The mammalian inner ear consists of the auditory and vestibular systems that are each composed of hair cells, supporting cells, neurons, mesenchymal cells, other epithelial cells, and blood vessels. We developed a novel protocol to sort auditory and vestibular tissues of newborn mouse inner ears into their major cellular components. Transcriptome profiling of the sorted cells identified cell type–specific expression clusters. Computational analysis detected transcription factors and microRNAs that play key roles in determining cell identity in the inner ear. Specifically, our analysis revealed the role of the Zeb1/miR-200b pathway in establishing epithelial and mesenchymal identity in the inner ear. Furthermore, we detected a misregulation of the ZEB1 pathway in the inner ear of Twirler mice, which manifest, among other phenotypes, malformations of the auditory and vestibular labyrinth. The association of misregulation of the ZEB1/miR-200b pathway with auditory and vestibular defects in the Twirler mutant mice uncovers a novel mechanism underlying deafness and balance disorders. Our approach can be employed to decipher additional complex regulatory networks underlying other hearing and balance mouse mutants. Author Summary The mammalian inner ear is a highly complex sensory organ, and mutations in more than 100 genes underlie hereditary human non-syndromic hearing loss. Nevertheless, little is known about the signaling cascades downstream of deafness genes. Genome-wide expression profiling is an invaluable tool for gaining systems-level understanding of biological processes. We developed and validated a simple and novel protocol to isolate sensory epithelial cells, neurons, blood vessels, and mesenchyme of auditory and vestibular epithelia from newborn wild-type mice. Our protocol is based on flow cytometry to sort and capture cells labeled with commercially available antibodies to endogenously expressed cluster of differentiation (CD) antigens. Using this strategy, we identified Zeb1 and miR200b as regulators of epithelial and mesenchymal identity in the mouse inner ear, and we further identified the signaling pathway disrupted by the Zeb1 mutation in the Twirler mouse mutant. We also show the utility of this approach for characterizing compartment-specific genes and protein–protein networks. Implementation of this isolation strategy to study other mouse mutants with hearing and balance phenotypes could overcome many of the obstacles to understanding the function of deafness genes. |
| Databáze: | OpenAIRE |
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