R.H. and D.C.-R. contributed equally to this work.
A model for mammalian cochlear hair cell differentiation in vitro: effects of retinoic acid on cytoskeletal proteins and potassium conductances
Article first published online: 26 FEB 2007
European Journal of Neuroscience
Volume 25, Issue 4, pages 957–973, February 2007
How to Cite
Helyer, R., Cacciabue-Rivolta, D., Davies, D., Rivolta, M. N., Kros, C. J. and Holley, M. C. (2007), A model for mammalian cochlear hair cell differentiation in vitro: effects of retinoic acid on cytoskeletal proteins and potassium conductances. European Journal of Neuroscience, 25: 957–973. doi: 10.1111/j.1460-9568.2007.05338.x
- Issue published online: 26 FEB 2007
- Article first published online: 26 FEB 2007
- Received 21 September 2006, revised 25 November 2006, accepted 30 November 2006
- cell line;
- organ of Corti
We have established a model for the in-vitro differentiation of mouse cochlear hair cells and have used it to explore the influence of retinoic acid on proliferation, cytoskeletal proteins and voltage-gated potassium conductances. The model is based on the conditionally immortal cell line University of Sheffield/ventral otocyst-epithelial cell line clone 36 (US/VOT-E36), derived from ventral otic epithelial cells of the mouse at embryonic day 10.5 and transfected with a reporter for myosin VIIa. Retinoic acid did not increase cell proliferation but led to up-regulation of myosin VIIa and formation of prominent actin rings that gave rise to numerous large, linear actin bundles. Cells expressing myosin VIIa had larger potassium conductances and did not express the cyclin-dependent kinase inhibitor p27kip1. US/VOT-E36 endogenously expressed the voltage-gated potassium channel α-subunits Kv1.3 and Kv2.1, which we subsequently identified in embryonic and neonatal hair cells in both auditory and vestibular sensory epithelia in vivo. These subunits could underlie the embryonic and neonatal delayed-rectifiers recorded in nascent hair cells in vivo. Kv2.1 was particularly prominent on the basolateral membrane of cochlear inner hair cells. Kv1.3 was distributed throughout all hair cells but tended to be localized to the cuticular plates. US/VOT-E36 recapitulates a coherent pattern of cell differentiation under the influence of retinoic acid and will provide a convenient model for screening the effects of other extrinsic factors on the differentiation of cochlear epithelial cell types in vitro.