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The Journal of Neuroscience, November 26, 2003, 23(34):10815-10826

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Cellular/Molecular
Developmental Assembly of Transduction Apparatus in Chick Basilar Papilla

Fan Si,¹ Hilary Brodie,¹ Peter G. Gillespie,² Ana E. Vazquez,¹ and Ebenezer N. Yamoah¹

¹Center for Neuroscience, Department of Otolaryngology, University of California, Davis, Davis, California 95616, and ²Oregon Hearing Research Center and Vollum Institute, Oregon Health and Science University, Portland, Oregon 97239

Hair cells, the sensory receptors of auditory and vestibular systems, use a transducer apparatus that renders them remarkably sensitive to mechanical displacement as minute as 1 nm. To study the embryonic development of the transducer apparatus in hair cells of the chick auditory papilla, we examined hair cells that have been labeled with N-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl) pyridiniumdibromide, which has been shown to permeate the transducer channels. In addition, mechanotransduction currents were recorded directly using whole-cell patch-clamp techniques. The structure of the hair bundle was examined using scanning electron microscopy, and immunofluorescence labeling for myosin 1c, myosin 7a, and plasma membrane Ca²⁺ ATPase 2 was studied to determine the developmental expression of these proteins in embryonic chick papillas. We demonstrate that the transducer apparatus is assembled jointly at embryonic day 11 (E11) of the developing chick basilar papilla. The resting open probability of the transducer channels was high at E12 (0.5) and remained substantially elevated at E14-16; it then declined to the mature value of 0.15 at E21. The displacement sensitivity of the transduction apparatus, the gating force, increased from E12 to E21. Although the expression of different components of the transducer apparatus and the transduction current peaked at E14-16, marked refinement occurred beyond E16. For example, myosin 1c appeared diffusely localized in hair bundles from E12 to E16, but subsequently consolidated into punctate pattern. The fine temporal and precise spatial assembly of the transducer apparatus likely contributes toward the exquisite sensitivity of the transduction ensemble.

Key words: hearing; inner ear; sensory receptors; myosin; PMCA; voltage clamp

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Received Aug 13, 2003; revised September 29, 2003; accepted October 6, 2003.

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