Abstract
Sensory hair cells in the mammalian cochlea convert mechanical stimuli into electrical impulses that subserve audition1,2. Loss of hair cells and their innervating neurons is the most frequent cause of hearing impairment3. Atonal homologue 1 (encoded by Atoh1, also known as Math1) is a basic helix–loop–helix transcription factor required for hair-cell development4,5,6, and its misexpression in vitro7,8 and in vivo9,10 generates hair-cell-like cells. Atoh1-based gene therapy to ameliorate auditory10 and vestibular11 dysfunction has been proposed. However, the biophysical properties of putative hair cells induced by Atoh1 misexpression have not been characterized. Here we show that in utero gene transfer of Atoh1 produces functional supernumerary hair cells in the mouse cochlea. The induced hair cells display stereociliary bundles, attract neuronal processes and express the ribbon synapse marker carboxy-terminal binding protein 2 (refs 12,13). Moreover, the hair cells are capable of mechanoelectrical transduction1,2 and show basolateral conductances with age-appropriate specializations. Our results demonstrate that manipulation of cell fate by transcription factor misexpression produces functional sensory cells in the postnatal mammalian cochlea. We expect that our in utero gene transfer paradigm will enable the design and validation of gene therapies to ameliorate hearing loss in mouse models of human deafness14,15.
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Acknowledgements
We thank C. Bresee and J. Jungwirth for expert technical support; C. Cepko and G. Nolan for plasmids EF1α–GFP and BMN–IRES–GFP, respectively; A. Kiernan, D. Fekete, S. Heller, A. Nguyen-Huynh, C. Bresee and J. Jungwirth for critical comments that improved the manuscript; D. Trune, B. Fritzsch and N. Segil for helpful discussions; S. Griest for statistical analyses; and M. Campbell and S. Nigra for exceptional animal care. This study was supported by grants from the National Institute on Deafness and Other Communication Disorders (J.V.B. and A.J.R.), the McKnight Endowment Fund for Neuroscience (J.J.B.) and the American Otological Society (Research Training Fellowship to S.P.G.).
Author Contributions The project was conceived by J.V.B. Experiments were planned and performed by S.P.G. and D.W.W. with advice from J.V.B., and were analysed by S.P.G., D.W.W. and J.V.B. J.V.B. performed the experimental embryology. A.J.R. conducted the electrophysiology experiments and interpreted the results. J.C.M. acquired the scanning electron micrographs. J.V.B. and A.J.R. wrote the paper.
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This file contains Supplementary Figures 1-4; Supplementary Tables 1 and 2; Supplementary Video Legends 1-4; and a discussion of the efficacy of transuterine microinjection and in vivo electroporation. (PDF 540 kb)
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Gubbels, S., Woessner, D., Mitchell, J. et al. Functional auditory hair cells produced in the mammalian cochlea by in utero gene transfer. Nature 455, 537–541 (2008). https://doi.org/10.1038/nature07265
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DOI: https://doi.org/10.1038/nature07265
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