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Differentiation of mammalian vestibular hair cells from conditionally immortal, postnatal supporting cells

Lawlor, P., Marcotti, W., Rivolta, M.N., Kros, C.J. and Holley, M.C. (1999) Differentiation of mammalian vestibular hair cells from conditionally immortal, postnatal supporting cells. The Journal of Neuroscience, 19 (21). pp. 9445-9458. ISSN 1529-2401

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Abstract

We provide evidence from a newly established, conditionally immortal cell line (UB/UE-1) that vestibular supporting cells from the mammalian inner ear can differentiate postnatally into more than one variant of hair cell. A clonal supporting cell line was established from pure utricular sensory epithelia of H2kbtsA58 transgenic mice 2 d after birth. Cell proliferation was dependent on conditional expression of the immortalizing gene, the “T” antigen from the SV40 virus. Proliferating cells expressed cytokeratins, and patch-clamp recordings revealed that they all expressed small membrane currents with little time-dependence. They stopped dividing within 2 d of being transferred to differentiating conditions, and within a week they formed three defined populations expressing membrane currents characteristic of supporting cells and two kinds of neonatal hair cell. The cells expressed several characteristic features of normal hair cells, including the transcription factor Brn3.1, a functional acetylcholine receptor composed of a9 subunits, and the cytoskeletal proteins myosin VI, myosin VIIa, and fimbrin. Immunofluorescence labeling and electron microscopy showed that the cells formed complex cytoskeletal arrays on their upper surfaces with structural features resembling those at the apices of normal hair cells. The cell line UB/UE-1 provides a valuable in vitro preparation in which the expression of numerous structural and physiological components can be initiated or upregulated during early stages of mammalian hair cell commitment and differentiation.

Item Type: Article
Copyright, Publisher and Additional Information: © 1999 The Society for Neuroscience. Reproduced in accordance with the publisher's self-archiving policy.
Keywords: mouse, vestibular, utricle, hair cells, epithelial cells, conditional immortalization, tsA58, differentiation, development, potassium current, inward rectifier
Institution: The University of Sheffield
Academic Units: The University of Sheffield > Faculty of Science (Sheffield) > School of Biological Sciences (Sheffield) > Department of Biomedical Science (Sheffield)
Depositing User: Ms Suzannah Rockett
Date Deposited: 28 Aug 2008 18:24
Last Modified: 07 Jun 2014 17:12
Published Version: http://www.jneurosci.org/cgi/reprint/19/21/9445
Status: Published
Publisher: HighWire
Refereed: Yes
URI: http://eprints.whiterose.ac.uk/id/eprint/4466

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