Identification of genetic and chemical modulators of zebrafish mechanosensory hair cell death
Kelly N Owens, Felipe Santos, Brock Roberts, Tor Linbo, Allison B Coffin, Anna J Knisely, Julian A Simon, Edwin W Rubel, David W Raible, Kelly N Owens, Felipe Santos, Brock Roberts, Tor Linbo, Allison B Coffin, Anna J Knisely, Julian A Simon, Edwin W Rubel, David W Raible
Abstract
Inner ear sensory hair cell death is observed in the majority of hearing and balance disorders, affecting the health of more than 600 million people worldwide. While normal aging is the single greatest contributor, exposure to environmental toxins and therapeutic drugs such as aminoglycoside antibiotics and antineoplastic agents are significant contributors. Genetic variation contributes markedly to differences in normal disease progression during aging and in susceptibility to ototoxic agents. Using the lateral line system of larval zebrafish, we developed an in vivo drug toxicity interaction screen to uncover genetic modulators of antibiotic-induced hair cell death and to identify compounds that confer protection. We have identified 5 mutations that modulate aminoglycoside susceptibility. Further characterization and identification of one protective mutant, sentinel (snl), revealed a novel conserved vertebrate gene. A similar screen identified a new class of drug-like small molecules, benzothiophene carboxamides, that prevent aminoglycoside-induced hair cell death in zebrafish and in mammals. Testing for interaction with the sentinel mutation suggests that the gene and compounds may operate in different pathways. The combination of chemical screening with traditional genetic approaches is a new strategy for identifying drugs and drug targets to attenuate hearing and balance disorders.
Conflict of interest statement
The authors have declared that no competing interests exist.
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References
- Gates GA, Couropmitree NN, Myers RH. Genetic associations in age-related hearing thresholds. Arch Otolaryngol Head Neck Surg. 1999;125:654–659.
- Johnson KR, Zheng QY, Noben-Trauth K. Strain background effects and genetic modifiers of hearing in mice. Brain Res. 2006;1091:79–88.
- Lanvers-Kaminsky C, Krefeld B, Dinnesen AG, Deuster D, Seifert E, et al. Continuous or repeated prolonged cisplatin infusions in children: a prospective study on ototoxicity, platinum concentrations, and standard serum parameters. Pediatr Blood Cancer. 2006;47:183–193.
- Nelson EG, Hinojosa R. Presbycusis: a human temporal bone study of individuals with downward sloping audiometric patterns of hearing loss and review of the literature. Laryngoscope. 2006;116:1–12.
- Sill AM, Stick MJ, Prenger VL, Phillips SL, Boughman JA, et al. Genetic epidemiologic study of hearing loss in an adult population. Am J Med Genet. 1994;54:149–153.
- Friedman TB, Griffith AJ. Human nonsyndromic sensorineural deafness. Annu Rev Genomics Hum Genet. 2003;4:341–402.
- Inoue H, Tanizawa Y, Wasson J, Behn P, Kalidas K, et al. A gene encoding a transmembrane protein is mutated in patients with diabetes mellitus and optic atrophy (Wolfram syndrome). Nat Genet. 1998;20:143–148.
- Lynch ED, Lee MK, Morrow JE, Welcsh PL, Leon PE, et al. Nonsyndromic deafness DFNA1 associated with mutation of a human homolog of the Drosophila gene diaphanous. Science. 1997;278:1315–1318.
- McGuirt WT, Prasad SD, Griffith AJ, Kunst HP, Green GE, et al. Mutations in COL11A2 cause non-syndromic hearing loss (DFNA13). Nat Genet. 1999;23:413–419.
- Forge A, Schacht J. Aminoglycoside antibiotics. Audiol Neurootol. 2000;5:3–22.
- Rybak LP, Whitworth CA, Mukherjea D, Ramkumar V. Mechanisms of cisplatin-induced ototoxicity and prevention. Hear Res. 2007;226:157–167.
- Nakashima T, Teranishi M, Hibi T, Kobayashi M, Umemura M. Vestibular and cochlear toxicity of aminoglycosides–a review. Acta Otolaryngol. 2000;120:904–911.
- Versnel H, Agterberg MJ, de Groot JC, Smoorenburg GF, Klis SF. Time course of cochlear electrophysiology and morphology after combined administration of kanamycin and furosemide. Hear Res. 2007;231:1–12.
- Prezant TR, Agapian JV, Bohlman MC, Bu X, Oztas S, et al. Mitochondrial ribosomal RNA mutation associated with both antibiotic-induced and non-syndromic deafness. Nat Genet. 1993;4:289–294.
- Guan MX, Fischel-Ghodsian N, Attardi G. Biochemical evidence for nuclear gene involvement in phenotype of non-syndromic deafness associated with mitochondrial 12S rRNA mutation. Hum Mol Genet. 1996;5:963–971.
- Guan MX, Fischel-Ghodsian N, Attardi G. A biochemical basis for the inherited susceptibility to aminoglycoside ototoxicity. Hum Mol Genet. 2000;9:1787–1793.
- Cheng AG, Cunningham LL, Rubel EW. Hair cell death in the avian basilar papilla: characterization of the in vitro model and caspase activation. J Assoc Res Otolaryngol. 2003;4:91–105.
- Dambly-Chaudiere C, Sapede D, Soubiran F, Decorde K, Gompel N, et al. The lateral line of zebrafish: a model system for the analysis of morphogenesis and neural development in vertebrates. Biol Cell. 2003;95:579–587.
- Nicolson T. The genetics of hearing and balance in zebrafish. Annu Rev Genet. 2005;39:9–22.
- Whitfield TT. Zebrafish as a model for hearing and deafness. J Neurobiol. 2002;53:157–171.
- Harris JA, Cheng AG, Cunningham LL, MacDonald G, Raible DW, et al. Neomycin-induced hair cell death and rapid regeneration in the lateral line of zebrafish (Danio rerio). J Assoc Res Otolaryngol. 2003;4:219–234.
- Murakami SL, Cunningham LL, Werner LA, Bauer E, Pujol R, et al. Developmental differences in susceptibility to neomycin-induced hair cell death in the lateral line neuromasts of zebrafish (Danio rerio). Hear Res. 2003;186:47–56.
- Santos F, MacDonald G, Rubel EW, Raible DW. Lateral line hair cell maturation is a determinant of aminoglycoside susceptibility in zebrafish (Danio rerio). Hear Res. 2006;213:25–33.
- Williams JA, Holder N. Cell turnover in neuromasts of zebrafish larvae. Hear Res. 2000;143:171–181.
- Ou HC, Raible DW, Rubel EW. Cisplatin-induced hair cell loss in zebrafish (Danio rerio) lateral line. Hear Res. 2007;233:46–53.
- Ton C, Parng C. The use of zebrafish for assessing ototoxic and otoprotective agents. Hear Res. 2005;208:79–88.
- Solnica-Krezel L, Schier AF, Driever W. Efficient recovery of ENU-induced mutations from the zebrafish germline. Genetics. 1994;136:1401–1420.
- Lister JA, Robertson CP, Lepage T, Johnson SL, Raible DW. nacre encodes a zebrafish microphthalmia-related protein that regulates neural-crest-derived pigment cell fate. Development. 1999;126:3757–3767.
- Balak KJ, Corwin JT, Jones JE. Regenerated hair cells can originate from supporting cell progeny: evidence from phototoxicity and laser ablation experiments in the lateral line system. J Neurosci. 1990;10:2502–2512.
- Bereiter-Hahn J. Dimethylaminostyrylmethylpyridiniumiodine (daspmi) as a fluorescent probe for mitochondria in situ. Biochim Biophys Acta. 1976;423:1–14.
- Shimoda N, Knapik EW, Ziniti J, Sim C, Yamada E, et al. Zebrafish genetic map with 2000 microsatellite markers. Genomics. 1999;58:219–232.
- Nalefski EA, Falke JJ. The C2 domain calcium-binding motif: structural and functional diversity. Protein Sci. 1996;5:2375–2390.
- Marcotti W, van Netten SM, Kros CJ. The aminoglycoside antibiotic dihydrostreptomycin rapidly enters mouse outer hair cells through the mechano-electrical transducer channels. J Physiol. 2005;567:505–521.
- Richardson GP, Forge A, Kros CJ, Fleming J, Brown SD, et al. Myosin VIIA is required for aminoglycoside accumulation in cochlear hair cells. J Neurosci. 1997;17:9506–9519.
- Seiler C, Nicolson T. Defective calmodulin-dependent rapid apical endocytosis in zebrafish sensory hair cell mutants. J Neurobiol. 1999;41:424–434.
- Nishikawa S, Sasaki F. Internalization of styryl dye FM1-43 in the hair cells of lateral line organs in Xenopus larvae. J Histochem Cytochem. 1996;44:733–741.
- Gale JE, Marcotti W, Kennedy HJ, Kros CJ, Richardson GP. FM1-43 dye behaves as a permeant blocker of the hair-cell mechanotransducer channel. J Neurosci. 2001;21:7013–7025.
- Meyers JR, MacDonald RB, Duggan A, Lenzi D, Standaert DG, et al. Lighting up the senses: FM1-43 loading of sensory cells through nonselective ion channels. J Neurosci. 2003;23:4054–4065.
- Steyger PS, Peters SL, Rehling J, Hordichok A, Dai CF. Uptake of gentamicin by bullfrog saccular hair cells in vitro. J Assoc Res Otolaryngol. 2003;4:565–578.
- Cunningham LL, Cheng AG, Rubel EW. Caspase activation in hair cells of the mouse utricle exposed to neomycin. J Neurosci. 2002;22:8532–8540.
- Cunningham LL, Matsui JI, Warchol ME, Rubel EW. Overexpression of Bcl-2 prevents neomycin-induced hair cell death and caspase-9 activation in the adult mouse utricle in vitro. J Neurobiol. 2004;60:89–100.
- Sugahara K, Rubel EW, Cunningham LL. JNK signaling in neomycin-induced vestibular hair cell death. Hear Res. 2006;221:128–135.
- Owens KN, Cunningham DE, Macdonald G, Rubel EW, Raible DW, et al. Ultrastructural analysis of aminoglycoside-induced hair cell death in the zebrafish lateral line reveals an early mitochondrial response. J Comp Neurol. 2007;502:522–543.
- Wu WJ, Sha SH, Schacht J. Recent advances in understanding aminoglycoside ototoxicity and its prevention. Audiol Neurootol. 2002;7:171–174.
- Avidor-Reiss T, Maer AM, Koundakjian E, Polyanovsky A, Keil T, et al. Decoding cilia function: defining specialized genes required for compartmentalized cilia biogenesis. Cell. 2004;117:527–539.
- Blacque OE, Perens EA, Boroevich KA, Inglis PN, Li C, et al. Functional genomics of the cilium, a sensory organelle. Curr Biol. 2005;15:935–941.
- North TE, Goessling W, Walkley CR, Lengerke C, Kopani KR, et al. Prostaglandin E2 regulates vertebrate haematopoietic stem cell homeostasis. Nature. 2007;447:1007–1011.
- Critchfield JW, Coligan JE, Folks TM, Butera ST. Casein kinase II is a selective target of HIV-1 transcriptional inhibitors. Proc Natl Acad Sci U S A. 1997;94:6110–6115.
- Gualberto A, Marquez G, Carballo M, Youngblood GL, Hunt SW, III, et al. p53 transactivation of the HIV-1 long terminal repeat is blocked by PD 144795, a calcineurin-inhibitor with anti-HIV properties. J Biol Chem. 1998;273:7088–7093.
- Wang D, Westerheide SD, Hanson JL, Baldwin AS., Jr Tumor necrosis factor alpha-induced phosphorylation of RelA/p65 on Ser529 is controlled by casein kinase II. J Biol Chem. 2000;275:32592–32597.
- Weinshilboum RM, Wang L. Pharmacogenetics and pharmacogenomics: development, science, and translation. Annu Rev Genomics Hum Genet. 2006;7:223–245.
- Humbert R, Adler DA, Disteche CM, Hassett C, Omiecinski CJ, et al. The molecular basis of the human serum paraoxonase activity polymorphism. Nat Genet. 1993;3:73–76.
- Rieder MJ, Reiner AP, Gage BF, Nickerson DA, Eby CS, et al. Effect of VKORC1 haplotypes on transcriptional regulation and warfarin dose. N Engl J Med. 2005;352:2285–2293.
- Westerfield M. Eugene (Oregon): University of Oregon Press; 2000. The Zebrafish book. A guide for the laboratory use of zebrafish (Danio rerio).
- Wikler MA. Clinical and Laboratory Standards Institute; 2006. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically. pp. 1–64.
- Wikler MA. Clinical and Laboratory Standards Institute; 2007. Performance Standards for Antimicrobial Susceptibility Testing. pp. 1–182.
- Chenna R, Sugawara H, Koike T, Lopez R, Gibson TJ, et al. Multiple sequence alignment with the Clustal series of programs. Nucleic Acids Res. 2003;31:3497–3500.
- Felsenstein J. PHLYIP–Phlylogeny Interference Package (version 3.2). Cladistics. 1989;5:164–166.
- Page RD. TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci. 1996;12:357–358.
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