The transmembrane inner ear (Tmie) protein is essential for normal hearing and balance in the zebrafish

December 15, 2009
106 (50) 21347-21352

Abstract

Little is known about the proteins that mediate mechanoelectrical transduction, the process by which acoustic and accelerational stimuli are transformed by hair cells of the inner ear into electrical signals. In our search for molecules involved in mechanotransduction, we discovered a line of deaf and uncoordinated zebrafish with defective hair-cell function. The hair cells of mutant larvae fail to incorporate fluorophores that normally traverse the transduction channels and their ears lack microphonic potentials in response to vibratory stimuli. Hair cells in the posterior lateral lines of mutants contain numerous lysosomes and have short, disordered hair bundles. Their stereocilia lack two components of the transduction apparatus, tip links and insertional plaques. Positional cloning revealed an early frameshift mutation in tmie, the zebrafish ortholog of the mammalian gene transmembrane inner ear. The mutant line therefore affords us an opportunity to investigate the role of the corresponding protein in mechanoelectrical transduction.

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Acknowledgments.

The authors thank H. Baier for Tg(pou4f3:gap43-mGFP) zebrafish, D. Gilmour for Tg(cldnB:lynGFP) zebrafish, V. Korzh for Et(krt4:GFP)sqet4 zebrafish, and J. Y. Kuwada for Tg(Hsp70:eGFP) zebrafish. We also thank A. Afolalu for excellent zebrafish husbandry and B. Fabella for computer programming and technical assistance with microphonic recordings. Members of our research group provided valuable comments on the manuscript. This investigation was supported by grants DC00241 and GM07739 from the National Institutes of Health and by grant SAF2006–04684 from the Ministerio de Ciencia e Innovación of Spain, of which H.L.-S. is a Ramón y Cajal Investigator. A.N. is the recipient of a Ruth L. Kirschstein National Research Service Award Predoctoral Fellowship. M.R.G. is an associate and A.J.H. is an investigator of the Howard Hughes Medical Institute.

References

1
AJ Hudspeth, How the ear's works work. Nature 341, 397–404 (1989).
2
C Petit, J Levilliers, JP Hardelin, Molecular genetics of hearing loss. Annu Rev Genet 35, 589–646 (2001).
3
K Dooley, LI Zon, Zebrafish: A model system for the study of human disease. Curr Opin Genet Dev 10, 252–256 (2000).
4
C Haddon, J Lewis, Early ear development in the embryo of the zebrafish, danio rerio. J Comp Neurol 365, 113–128 (1996).
5
JHS Blaxter, LA Fuiman The Mechanosensory Lateral Line: Neurobiology and Evolution, eds S Coombs, P Görner, H Münz (Springer, New York), pp. 481–499 (1989).
6
Y Asai, et al., Mutation of the atrophin2 gene in the zebrafish disrupts signaling by fibroblast growth factor during development of the inner ear. Proc Natl Acad Sci USA 103, 9069–9074 (2006).
7
P Haffter, C Nusslein-Volhard, Large scale genetics in a small vertebrate, the zebrafish. Int J Dev Biol 40, 221–227 (1996).
8
JA Kappler, CJ Starr, DK Chan, R Kollmar, AJ Hudspeth, A nonsense mutation in the gene encoding a zebrafish myosin vi isoform causes defects in hair-cell mechanotransduction. Proc Natl Acad Sci USA 101, 13056–13061 (2004).
9
CJ Starr, JA Kappler, DK Chan, R Kollmar, AJ Hudspeth, Mutation of the zebrafish choroideremia gene encoding rab escort protein 1 devastates hair cells. Proc Natl Acad Sci USA 101, 2572–2577 (2004).
10
SR Blechinger, et al., The heat-inducible zebrafish hsp70 gene is expressed during normal lens development under non-stress conditions. Mech Dev 112, 213–215 (2002).
11
MB Foreman, RC Eaton, The direction change concept for reticulospinal control of goldfish escape. J Neurosci 13, 4101–4113 (1993).
12
JE Gale, W Marcotti, HJ Kennedy, CJ Kros, GP Richardson, FM1–43 dye behaves as a permeant blocker of the hair-cell mechanotransducer channel. J Neurosci 21, 7013–7025 (2001).
13
JR Meyers, et al., Lighting up the senses: FM1–43 loading of sensory cells through nonselective ion channels. J Neurosci 23, 4054–4065 (2003).
14
T Xiao, T Roeser, W Staub, H Baier, A gfp-based genetic screen reveals mutations that disrupt the architecture of the zebrafish retinotectal projection. Development 132, 2955–2967 (2005).
15
T Nicolson, et al., Genetic analysis of vertebrate sensory hair cell mechanosensation: The zebrafish circler mutants. Neuron 20, 271–283 (1998).
16
P Haas, D Gilmour, Chemokine signaling mediates self-organizing tissue migration in the zebrafish lateral line. Dev Cell 10, 673–680 (2006).
17
H López-Schier, A Hudspeth, A two-step mechanism underlies the planar polarization of regenerating sensory hair cells. Proc Natl Acad Sci USA 103, 18615–18620 (2006).
18
PP Hernandez, FA Olivari, AF Sarrazin, PC Sandoval, ML Allende, Regeneration in zebrafish lateral line neuromasts: Expression of the neural progenitor cell marker sox2 and proliferation-dependent and-independent mechanisms of hair cell renewal. Dev Neurobiol 67, 637–654 (2007).
19
YC Shen, et al., The transmembrane inner ear (tmie) gene contributes to vestibular and lateral line development and function in the zebrafish (Danio rerio). Dev Dyn 237, 941–952 (2008).
20
T Hirokawa, S Boon-Chieng, S Mitaku, Sosui: Classification and secondary structure prediction system for membrane proteins. Bioinformatics 14, 378–379 (1998).
21
KL Mitchem, et al., Mutation of the novel gene tmie results in sensory cell defects in the inner ear of spinner, a mouse model of human hearing loss dfnb6. Hum Mol Genet 11, 1887–1898 (2002).
22
WH Chung, et al., Cochlear pathology of the circling mouse: A new mouse model of dfnb6. Acta Otolaryngol 127, 244–251 (2007).
23
S Naz, et al., Mutations in a novel gene, tmie, are associated with hearing loss linked to the dfnb6 locus. Am J Hum Genet 71, 632–636 (2002).
24
RL Santos, et al., Novel sequence variants in the tmie gene in families with autosomal recessive nonsyndromic hearing impairment. J Mol Med 84, 226–231 (2006).
25
Y Saheki, CI Bargmann, Presynaptic CaV2 calcium channel traffic requires CALF-1 and the α2δ subunit UNC-36. Nat Neurosci 12, 1257–1265 (2009).
26
M Westerfield The Zebrafish Book. A Guide for the Laboratory Use of Zebrafish (Danio rerio) (University of Oregon Press, Ed 4, Eugene, 2000).
27
JH Postlethwait, WS Talbot, Zebrafish genomics: From mutants to genes. Trends Genet 13, 183–190 (1997).
28
A Nagiel, D Andor-Ardo, AJ Hudspeth, Specificity of afferent synapses onto plane-polarized hair cells in the posterior lateral line of the zebrafish. J Neurosci 28, 8442–8453 (2008).
29
R Tang, A Dodd, D Lai, WC McNabb, DR Love, Validation of zebrafish (Danio rerio) reference genes for quantitative real-time rt-pcr normalization. Acta Biochim Biophys Sin (Shanghai) 39, 384–390 (2007).

Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 106 | No. 50
December 15, 2009
PubMed: 19934034

Classifications

Submission history

Received: September 3, 2009
Published online: December 15, 2009
Published in issue: December 15, 2009

Keywords

  1. auditory system
  2. hair cell
  3. lateral line
  4. mechanoelectrical transduction
  5. vestibular system

Acknowledgments

The authors thank H. Baier for Tg(pou4f3:gap43-mGFP) zebrafish, D. Gilmour for Tg(cldnB:lynGFP) zebrafish, V. Korzh for Et(krt4:GFP)sqet4 zebrafish, and J. Y. Kuwada for Tg(Hsp70:eGFP) zebrafish. We also thank A. Afolalu for excellent zebrafish husbandry and B. Fabella for computer programming and technical assistance with microphonic recordings. Members of our research group provided valuable comments on the manuscript. This investigation was supported by grants DC00241 and GM07739 from the National Institutes of Health and by grant SAF2006–04684 from the Ministerio de Ciencia e Innovación of Spain, of which H.L.-S. is a Ramón y Cajal Investigator. A.N. is the recipient of a Ruth L. Kirschstein National Research Service Award Predoctoral Fellowship. M.R.G. is an associate and A.J.H. is an investigator of the Howard Hughes Medical Institute.

Authors

Affiliations

Michelle R. Gleason
Howard Hughes Medical Institute and Laboratory of Sensory Neuroscience, The Rockefeller University, New York, NY 10065; and
Aaron Nagiel
Howard Hughes Medical Institute and Laboratory of Sensory Neuroscience, The Rockefeller University, New York, NY 10065; and
Sophie Jamet
Laboratory of Sensory Cell Biology and Organogenesis, Centre de Regulació Genòmica-Barcelona Biomedical Research Park (PRBB), 08003 Barcelona, Spain
Present address: UMR 7102 CNRS, Université Pierre et Marie Curie, 9, Quai St Bernard, 75005 Paris, France.
Maria Vologodskaia
Howard Hughes Medical Institute and Laboratory of Sensory Neuroscience, The Rockefeller University, New York, NY 10065; and
Hernán López-Schier2 [email protected]
Howard Hughes Medical Institute and Laboratory of Sensory Neuroscience, The Rockefeller University, New York, NY 10065; and
Laboratory of Sensory Cell Biology and Organogenesis, Centre de Regulació Genòmica-Barcelona Biomedical Research Park (PRBB), 08003 Barcelona, Spain
A. J. Hudspeth2 [email protected]
Howard Hughes Medical Institute and Laboratory of Sensory Neuroscience, The Rockefeller University, New York, NY 10065; and

Notes

2
To whom correspondence may be addressed. E-mail: [email protected] or [email protected]
Contributed by A. J. Hudspeth, October 7, 2009
Author contributions: M.R.G., H.L.-S., and A.J.H. designed research; M.R.G., A.N., S.J., M.V., and H.L.-S. performed research; M.R.G. and H.L.-S. analyzed data; and M.R.G., A.N., S.J., M.V., H.L.-S., and A.J.H. wrote the paper.

Competing Interests

The authors declare no conflict of interest.

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    The transmembrane inner ear (Tmie) protein is essential for normal hearing and balance in the zebrafish
    Proceedings of the National Academy of Sciences
    • Vol. 106
    • No. 50
    • pp. 21009-21458

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