Abstract
There are different β-tubulin isoforms in microtubules of vertebrate tissues. However, their functional significance is still largely unknown. In the present study, we investigated the localization of five β-tubulin isotypes (β1–5) within the hearing organ during development in rat. By using confocal microscopy, we showed that with the exception of the β3-tubulin isoform that was specific to nerve fibres, all the different β-tubulin isoforms were mainly present in the supporting cells. Contrary to β1–4-tubulins, we also found that the β5-tubulin isoform appeared only at a key stage of the post-natal development in specific cell types (pillar cells and Deiters’ cells). By using transmission electron microscopy, we revealed further that this developmental stage coincided with the formation of two separate bundles of microtubules from a unique one in these supporting cells. Together, these results suggest that the β5-tubulin isoform might be involved in the generation of new microtubule bundles from a pre-existing one.
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Acknowledgments
We thank Mrs P. Piscicelli for her skilful technical assistance and Dr. Richard F. Ludueña (Department of Biochemistry, University of Texas Health Science Center at San Antonio, San Antonio, Texas) for generous gift of the antibody anti-β5-tubulin. We thank the GIGA-Imaging and Flow Cytometry platform for technical support. This work was supported by Fonds de la recherche scientifique (FNRS-FRS). J.R. and N.J. are PhD grant holders of the FRIA. M.C. was a FRS-FNRS Research Fellow.
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Justine Renauld and Nicolas Johnen have contributed equally to this article.
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418_2015_1350_MOESM1_ESM.jpg
Fig S9. Ultrastructure of Pillar’s cells at P4 (a-d) and P25 (e–f). (a) General view. Bar 2.5 μm. (b, c, d) Details of their cytoplasm with microtubule bundles starting from the apical part of the cell (arrowheads). Bar 250 nm. (e) General view. Bar 2.5 µm (f) An enlargement of two microtubule bundles arranged in two different directions (arrowheads). Ct tunnel of Corti Ip inner pillar cell, IHC inner hair cell, Ns space of Nuel, OHC outer hair cell, Op outer pillar cell. Bar 250 nm. (JPEG 1421 kb)
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Renauld, J., Johnen, N., Thelen, N. et al. Spatio-temporal dynamics of β-tubulin isotypes during the development of the sensory auditory organ in rat. Histochem Cell Biol 144, 403–416 (2015). https://doi.org/10.1007/s00418-015-1350-2
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DOI: https://doi.org/10.1007/s00418-015-1350-2