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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References
Akhshi TK, Wernike D, Piekny A (2014) Microtubules and actin crosstalk in cell migration and division. Cytoskeleton 71:1–23. doi:10.1002/cm.21150
Banerjee A, Jensen-Smith H, Lazzell A et al (2008) Localization of βv tubulin in the cochlea and cultured cells with a novel monoclonal antibody. Cell Motil Cytoskelet 65:505–514. doi:10.1002/cm.20280
Bhattacharya R, Cabral F (2004) A ubiquitous beta-tubulin disrupts microtubule assembly and inhibits cell proliferation. Mol Biol Cell 15:3123–3131. doi:10.1091/mbc.E04
Bhattacharya R, Yang H, Cabral F (2011) Class V β-tubulin alters dynamic instability and stimulates microtubule detachment from centrosomes. Mol Biol Cell 22:1025–1034. doi:10.1091/mbc.E10-10-0822
Bounoutas A, Hagan RO, Chalfie M (2009) The multipurpose 15-protofilament microtubules in C. elegans have specific roles in mechanosensation. Curr Biol 19:1362–1367. doi:10.1016/j.cub.2009.06.036
Cloes M, Renson T, Johnen N et al (2013) Differentiation of Boettcher’s cells during postnatal development of rat cochlea. Cell Tissue Res 354:707–716. doi:10.1007/s00441-013-1705-8
Cowan NJ, Lewis SA, Gu W, Burgoyne RD (1988) Tubulin isotypes and their interaction with microtubule associated proteins. Protoplasma 145:106–111. doi:10.1007/BF01349346
Etienne-Manneville S (2013) Microtubules in cell migration. Annu Rev Cell Dev Biol 29:471–499. doi:10.1146/annurev-cellbio-101011-155711
Fritzsch B, Pan N, Jahan I, Elliott KL (2014) Inner ear development: building a spiral ganglion and an organ of Corti out of unspecified ectoderm. Cell Tissue Res. doi:10.1007/s00441-014-2031-5
Fukushige T, Siddiqui ZK, Chou M et al (1999) MEC-12, an alpha-tubulin required for touch sensitivity in C. elegans. J Cell Sci 112(Pt 3):395–403
Fulton C, Simpson PA (1976) Selective synthesis and utilization of flagellar tubulin. The multi-tubulin hypothesis. Cell Motil 3:987–1005
Hallworth R, Ludueña RF (2000) Differential expression of beta tubulin isotypes in the adult gerbil cochlea. Hear Res 148:161–172. doi:10.1016/S0378-5955(00)00149-0
Hallworth R, McCoy M, Polan-Curtain J (2000) Tubulin expression in the developing and adult gerbil organ of Corti. Hear Res 139:31–41. doi:10.1016/S0378-5955(99)00165-3
Hari M, Wang Y, Veeraraghavan S, Cabral F (2003) Mutations in alpha- and beta-tubulin that stabilize microtubules and confer resistance to colcemid and vinblastine. Mol Cancer Ther 2:597–605
Henderson CG, Tucker JB, Chaplin MA et al (1994) Reorganization of the centrosome and associated microtubules during the morphogenesis of a mouse cochlear epithelial cell. J Cell Sci 107(Pt 2):589–600
Henderson CG, Tucker JB, Mogensen MM et al (1995) Three microtubule-organizing centres collaborate in a mouse cochlear epithelial cell during supracellularly coordinated control of microtubule positioning. J Cell Sci 108(Pt 1):37–50
Jensen-Smith HC, Eley J, Steyger PS et al (2003) Cell type-specific reduction of β tubulin isotypes synthesized in the developing gerbil organ of Corti. J Neurocytol 32:185–197. doi:10.1023/B:NEUR.0000005602.18713.02
Johnen N, Francart M-E, Thelen N et al (2012) Evidence for a partial epithelial–mesenchymal transition in postnatal stages of rat auditory organ morphogenesis. Histochem Cell Biol 138:477–488. doi:10.1007/s00418-012-0969-5
Joshi HC, Chu D, Buxbaum RE, Heidemann SR (1985) Tension and compression in the cytoskeleton of PC 12 neurites. J Cell Biol 101:697–705. doi:10.1083/jcb.101.3.697
Karavitaki KD, Mountain DC (2007) Evidence for outer hair cell driven oscillatory fluid flow in the tunnel of Corti. Biophys J 92:3284–3293. doi:10.1529/biophysj.106.084087
Lee HO, Norden C (2013) Mechanisms controlling arrangements and movements of nuclei in pseudostratified epithelia. Trends Cell Biol 23:141–150. doi:10.1016/j.tcb.2012.11.001
Lewis SA, Lee MG, Cowan NJ (1985) Five mouse tubulin isotypes and their regulated expression during development. J Cell Biol 101:852–861. doi:10.1083/jcb.101.3.852
Locher H, Frijns JHM, Huisman MA, de Sousa Lopes SMC (2013) TUBB3: neuronal marker or melanocyte mimic? Cell Transplant. doi:10.3727/096368913X674099
Ludueña RF (1998) Multiple forms of tubulin: different gene products and covalent modifications. Int Rev Cytol 178:207–275. doi:10.1016/S0074-7696(08)62138-5
Parsa A, Webster P, Kalinec F (2012) Deiters cells tread a narrow path—the Deiters cells-basilar membrane junction. Hear Res 290:13–20. doi:10.1016/j.heares.2012.05.006
Perry B, Jensen-Smith HC, Ludueña RF, Hallworth R (2003) Selective expression of beta tubulin isotypes in gerbil vestibular sensory epithelia and neurons. J Assoc Res Otolaryngol 4:329–338. doi:10.1007/s10162-002-2048-4
Raff EC (1997) Microtubule architecture specified by a beta-tubulin isoform. Science 275:70–73. doi:10.1126/science.275.5296.70
Roach MC, Boucher VL, Walss C et al (1998) Preparation of a monoclonal antibody specific for the class I isotype of β-tubulin: the β isotypes of tubulin differ in their cellular distributions within human tissues. Cell Motil Cytoskelet 39:273–285. doi:10.1002/(SICI)1097-0169(1998)39:4<273::AID-CM3>3.0.CO;2-4
Saillour Y, Broix L, Bruel-Jungerman E et al (2014) Beta tubulin isoforms are not interchangeable for rescuing impaired radial migration due to Tubb3 knockdown. Hum Mol Genet 23:1516–1526. doi:10.1093/hmg/ddt538
Savage C, Hamelin M, Culotti JG et al (1989) mec-7 is a beta-tubulin gene required for the production of 15-protofilament microtubules in Caenorhabditis elegans. Genes Dev 3:870–881. doi:10.1101/gad.3.6.870
Slepecky NB, Henderson CG, Saha S (1995) Post-translational modifications of tubulin suggest that dynamic microtubules are present in sensory cells and stable microtubules are present in supporting cells of the mammalian cochlea. Hear Res 91:136–147. doi:10.1016/0378-5955(95)00184-0
Souter M, Nevill G, Forge A (1997) Postnatal maturation of the organ of Corti in gerbils: morphology and physiological responses. J Comp Neurol 386:635–651. doi:10.1002/(SICI)1096-9861(19971006)386:4<635::AID-CNE9>3.0.CO;2-3
Steyger PS, Furness DN, Hackney CM, Richardson GP (1989) Tubulin and microtubules in cochlear hair cells: comparative immunocytochemistry and ultrastructure. Hear Res 42:1–16. doi:10.1016/0378-5955(89)90113-5
Szarama KB, Gavara N, Petralia RS et al (2012) Cytoskeletal changes in actin and microtubules underlie the developing surface mechanical properties of sensory and supporting cells in the mouse cochlea. Development (Cambridge, England) 139:2187–2197. doi:10.1242/dev.073734
Tannenbaum J, Slepecky NB (1997) Localization of microtubules containing posttranslationally modified tubulin in cochlear epithelial cells during development. Cell Motil Cytoskelet 38:146–162. doi:10.1002/(SICI)1097-0169(1997)38:2<146::AID-CM4>3.0.CO;2-5
Thelen N, Breuskin I, Malgrange B, Thiry M (2009) Early identification of inner pillar cells during rat cochlear development. Cell Tissue Res 337:1–14. doi:10.1007/s00441-009-0810-1
Tischfield MA, Baris HN, Wu C et al (2010) Human TUBB3 mutations perturb microtubule dynamics, kinesin interactions, and axon guidance. Cell 140:74–87. doi:10.1016/j.cell.2009.12.011
Tucker JB, Paton CC, Richardson GP et al (1992) A cell surface-associated centrosomal layer of microtubule-organizing material in the inner pillar cell of the mouse cochlea. J Cell Sci 102(Pt 2):215–226
Tucker JB, Mogensen MM, Henderson CG et al (1998) Nucleation and capture of large cell surface-associated microtubule arrays that are not located near centrosomes in certain cochlear epithelial cells. J Anat 192(Pt 1):119–130. doi:10.1046/j.1469-7580.1998.19210119.x
Wade RH (2009) On and around microtubules: an overview. Mol Biotechnol 43:177–191. doi:10.1007/s12033-009-9193-5
Yang H, Cabral F, Bhattacharya R (2009) Tubulin isotype specificity and identification of the epitope for antibody Tub 2.1. Protein Eng Des Sel 22:625–629. doi:10.1093/protein/gzp046
Zagadou BF, Mountain DC (2012) Analysis of the cochlear amplifier fluid pump hypothesis. J Assoc Res Otolaryngol 13:185–197. doi:10.1007/s10162-011-0308-x
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