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CD44 is a Marker for the Outer Pillar Cells in the Early Postnatal Mouse Inner Ear
Authors:Ronna Hertzano  Chandrakala Puligilla  Siaw-Lin Chan  Caroline Timothy  Didier A. Depireux  Zubair Ahmed  Jeffrey Wolf  David J. Eisenman  Thomas B. Friedman  Sheikh Riazuddin  Matthew W. Kelley  Scott E. Strome
Affiliation:1. Department of Otorhinolaryngology–Head and Neck Surgery, University of Maryland, 16 South Eutaw Street, Suite 500, Baltimore, MD, 21201, USA
2. Section on Developmental Neuroscience, National Institute on Deafness and other Communication Disorders, National Institutes of Health, Bethesda, MD, 20892, USA
3. Institute for Systems Research, School of Engineering, University of Maryland, College Park, MD, 20742, USA
4. Division of Pediatric Ophthalmology, Cincinnati Children’s Hospital Research Foundation, Cincinnati, OH, 45229, USA
5. Department of Ophthalmology, University of Cincinnati, Cincinnati, OH, 45229, USA
6. Section on Human Genetics, Laboratory of Molecular Genetics, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Rockville, MD, 20850, USA
7. National Centre of Excellence in Molecular Biology, Punjab University, Lahore, Pakistan
Abstract:Cluster of differentiation antigens (CD proteins) are classically used as immune cell markers. However, their expression within the inner ear is still largely undefined. In this study, we explored the possibility that specific CD proteins might be useful for defining inner ear cell populations. mRNA expression profiling of microdissected auditory and vestibular sensory epithelia revealed 107 CD genes as expressed in the early postnatal mouse inner ear. The expression of 68 CD genes was validated with real-time RT-PCR using RNA extracted from microdissected sensory epithelia of cochleae, utricles, saccules, and cristae of newborn mice. Specifically, CD44 was identified as preferentially expressed in the auditory sensory epithelium. Immunohistochemistry revealed that within the early postnatal organ of Corti, the expression of CD44 is restricted to outer pillar cells. In order to confirm and expand this finding, we characterized the expression of CD44 in two different strains of mice with loss- and gain-of-function mutations in Fgfr3 which encodes a receptor for FGF8 that is essential for pillar cell development. We found that the expression of CD44 is abolished from the immature pillar cells in homozygous Fgfr3 knockout mice. In contrast, both the outer pillar cells and the aberrant Deiters’ cells in the Fgfr3 P244R/ + mice express CD44. The deafness phenotype segregating in DFNB51 families maps to a linkage interval that includes CD44. To study the potential role of CD44 in hearing, we characterized the auditory system of CD44 knockout mice and sequenced the entire open reading frame of CD44 of affected members of DFNB51 families. Our results suggest that CD44 does not underlie the deafness phenotype of the DFNB51 families. Finally, our study reveals multiple potential new cell type-specific markers in the mouse inner ear and identifies a new marker for outer pillar cells.
Keywords:CD44   FGFR3   cochlea   outer pillar cells   deafness
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