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Delayed Otolith Development Does Not Impair Vestibular Circuit Formation in Zebrafish

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Abstract

What is the role of normally patterned sensory signaling in development of vestibular circuits? For technical reasons, including the difficulty in depriving animals of vestibular inputs, this has been a challenging question to address. Here we take advantage of a vestibular-deficient zebrafish mutant, rock solo AN66, in order to examine whether normal sensory input is required for formation of vestibular-driven postural circuitry. We show that the rock solo AN66 mutant is a splice site mutation in the secreted glycoprotein otogelin (otog), which we confirm through both whole genome sequencing and complementation with an otog early termination mutant. Using confocal microscopy, we find that elements of postural circuits are anatomically normal in rock solo AN66 mutants, including hair cells, vestibular ganglion neurons, and vestibulospinal neurons. Surprisingly, the balance and postural deficits that are readily apparent in younger larvae disappear around 2 weeks of age. We demonstrate that this behavioral recovery follows the delayed development of the anterior (utricular) otolith, which appears around 14 days post-fertilization (dpf), compared to 1 dpf in WT. These findings indicate that utricular signaling is not required for normal structural development of the inner ear and vestibular nucleus neurons. Furthermore, despite the otolith’s developmental delay until well after postural behaviors normally appear, downstream circuits can drive righting reflexes within ∼1–2 days of its arrival, indicating that vestibular circuit wiring is not impaired by a delay in patterned activity. The functional recovery of postural behaviors may shed light on why humans with mutations in otog exhibit only subclinical vestibular deficits.

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References

  • Bagnall MW, McLean DL (2014) Modular organization of axial microcircuits in zebrafish. Science 343:197–200

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Basaldella E, Takeoka A, Sigrist M, Arber S (2015) Multisensory signaling shapes vestibulo-motor circuit specificity. Cell 163:301–312

    Article  CAS  PubMed  Google Scholar 

  • Beck JC, Gilland E, Tank DW, Baker R (2004) Quantifying the ontogeny of optokinetic and vestibuloocular behaviors in zebrafish, medaka, and goldfish. J Neurophysiol 92:3546–3561

    Article  PubMed  Google Scholar 

  • Bever MM, Fekete DM (2002) Atlas of the developing inner ear in zebrafish. Developmental dynamics : an official publication of the American Association of Anatomists 223:536–543

    Article  Google Scholar 

  • Boyle R, Mensinger AF, Yoshida K, Usui S, Intravaia A, Tricas T, Highstein SM (2001) Neural readaptation to Earth’s gravity following return from space. J Neurophysiol 86:2118–2122

    CAS  PubMed  Google Scholar 

  • Branoner F, Straka H (2015) Semicircular canal-dependent developmental tuning of translational vestibulo-ocular reflexes in Xenopus laevis. Developmental neurobiology 75:1051–1067

    Article  CAS  PubMed  Google Scholar 

  • Caston J, Precht W, Blanks RHI (1977) Response characteristics of frog’s lagena afferents to natural stimulation. J Comp Physiol 118:273–289

    Article  Google Scholar 

  • Cingolani P, Platts A, Wang le L, Coon M, Nguyen T, Wang L, Land SJ, Lu X, Ruden DM (2012) A program for annotating and predicting the effects of single nucleotide polymorphisms, SnpEff: SNPs in the genome of Drosophila melanogaster strain w1118; iso-2; iso-3. Fly (Austin) 6:80–92

    Article  CAS  Google Scholar 

  • Cohen-Salmon M, El-Amraoui A, Leibovici M, Petit C (1997) Otogelin: a glycoprotein specific to the acellular membranes of the inner ear. Proc Natl Acad Sci U S A 94:14450–14455

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Eatock RA, Songer JE (2011) Vestibular hair cells and afferents: two channels for head motion signals. Annu Rev Neurosci 34:501–534

    Article  CAS  PubMed  Google Scholar 

  • El Hakam Kamareddin C, Magnol L, Blanquet V (2015) A new Otogelin ENU mouse model for autosomal-recessive nonsyndromic moderate hearing impairment. Springerplus 4:730

    Article  PubMed  PubMed Central  Google Scholar 

  • El-Amraoui A, Cohen-Salmon M, Petit C, Simmler MC (2001) Spatiotemporal expression of otogelin in the developing and adult mouse inner ear. Hear Res 158:151–159

    Article  CAS  PubMed  Google Scholar 

  • Elliott KL, Houston DW, DeCook R, Fritzsch B (2015) Ear manipulations reveal a critical period for survival and dendritic development at the single-cell level in Mauthner neurons. Developmental neurobiology 75:1339–1351

    Article  PubMed  PubMed Central  Google Scholar 

  • Eugene D, Deforges S, Vibert N, Vidal PP (2009) Vestibular critical period, maturation of central vestibular neurons, and locomotor control. Ann N Y Acad Sci 1164:180–187

    Article  PubMed  Google Scholar 

  • Fernandez C, Goldberg JM (1976) Physiology of peripheral neurons innervating otolith organs of the squirrel monkey. I. Response to static tilts and to long-duration centrifugal force. J Neurophysiol 39:970–984

    CAS  PubMed  Google Scholar 

  • Fritzsch B (1990) Experimental reorganization in the alar plate of the clawed toad, Xenopus laevis. I. Quantitative and qualitative effects of embryonic otocyst extirpation. Brain Res Dev Brain Res 51:113–122

    Article  CAS  PubMed  Google Scholar 

  • Goodyear RJ, Legan PK, Christiansen JR, Xia B, Korchagina J, Gale JE, Warchol ME, Corwin JT, Richardson GP (2010) Identification of the hair cell soma-1 antigen, HCS-1, as otoferlin. J Assoc Res Otolaryngol 11:573–586

    Article  PubMed  PubMed Central  Google Scholar 

  • Haddon C, Lewis J (1996) Early ear development in the embryo of the zebrafish, Danio rerio. J Comp Neurol 365:113–128

    Article  CAS  PubMed  Google Scholar 

  • Hensch TK (2004) Critical period regulation. Annu Rev Neurosci 27:549–579

    Article  CAS  PubMed  Google Scholar 

  • Holmdahl R, Malissen B (2012) The need for littermate controls. Eur J Immunol 42:45–47

    Article  CAS  PubMed  Google Scholar 

  • Hubel DH, Wiesel TN (1970) The period of susceptibility to the physiological effects of unilateral eye closure in kittens. J Physiol 206:419–436

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kettleborough RN, Busch-Nentwich EM, Harvey SA, Dooley CM, de Bruijn E, van Eeden F, Sealy I, White RJ, Herd C, Nijman IJ, Fenyes F, Mehroke S, Scahill C, Gibbons R, Wali N, Carruthers S, Hall A, Yen J, Cuppen E, Stemple DL (2013) A systematic genome-wide analysis of zebrafish protein-coding gene function. Nature 496:494–497

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Khorevin VI (2008) The lagena (the third otolith endorgan in vertebrates). Neurophysiology 40:142–159

    Article  Google Scholar 

  • Kimmel CB, Powell SL, Metcalfe WK (1982) Brain neurons which project to the spinal cord in young larvae of the zebrafish. J Comp Neurol 205:112–127

    Article  CAS  PubMed  Google Scholar 

  • Lambert FM, Beck JC, Baker R, Straka H (2008) Semicircular canal size determines the developmental onset of angular vestibuloocular reflexes in larval Xenopus. J Neurosci 28:8086–8095

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leake PA, Hradek GT, Chair L, Snyder RL (2006) Neonatal deafness results in degraded topographic specificity of auditory nerve projections to the cochlear nucleus in cats. J Comp Neurol 497:13–31

    Article  PubMed  PubMed Central  Google Scholar 

  • Levi-Montalcini R (1949) The development of the acoustico-vestibular centers in the chick embryo in the absence of the afferent root fibers and of descending fiber tracts. J Comp Neurol 91:209–241

    Article  CAS  PubMed  Google Scholar 

  • Li H (2014) Toward better understanding of artifacts in variant calling from high-coverage samples. Bioinformatics 30:2843–2851

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, Marth G, Abecasis G, Durbin R, Genome Project Data Processing S (2009) The sequence alignment/map format and SAMtools. Bioinformatics 25:2078–2079

    Article  PubMed  PubMed Central  Google Scholar 

  • Lu Z, Xu Z, Buchser WJ (2003) Acoustic response properties of lagenar nerve fibers in the sleeper goby, Dormitator latifrons. Journal of comparative physiology A, Neuroethology, sensory, neural, and behavioral physiology 189:889–905

    Article  CAS  PubMed  Google Scholar 

  • MacNaughtan IPJ, McNally WJ (1946) Some experiments which indicate that the frog’s lagena has an equilibrial function. J Laryngol Otol 61:204–214

    Article  CAS  PubMed  Google Scholar 

  • Mo W, Chen F, Nechiporuk A, Nicolson T (2010) Quantification of vestibular-induced eye movements in zebrafish larvae. BMC Neurosci 11:110

    Article  PubMed  PubMed Central  Google Scholar 

  • Moorman SJ, Cordova R, Davies SA (2002) A critical period for functional vestibular development in zebrafish. Developmental dynamics : an official publication of the American Association of Anatomists 223:285–291

    Article  Google Scholar 

  • Oonk AM, Leijendeckers JM, Huygen PL, Schraders M, del Campo M, del Castillo I, Tekin M, Feenstra I, Beynon AJ, Kunst HP, Snik AF, Kremer H, Admiraal RJ, Pennings RJ (2014) Similar phenotypes caused by mutations in OTOG and OTOGL. Ear Hear 35:e84–e91

    Article  PubMed  PubMed Central  Google Scholar 

  • Paffenholz R, Bergstrom RA, Pasutto F, Wabnitz P, Munroe RJ, Jagla W, Heinzmann U, Marquardt A, Bareiss A, Laufs J, Russ A, Stumm G, Schimenti JC, Bergstrom DE (2004) Vestibular defects in head-tilt mice result from mutations in Nox3, encoding an NADPH oxidase. Genes Dev 18:486–491

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pittman AJ, Law MY, Chien CB (2008) Pathfinding in a large vertebrate axon tract: isotypic interactions guide retinotectal axons at multiple choice points. Development 135:2865–2871

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Riley BB, Moorman SJ (2000) Development of utricular otoliths, but not saccular otoliths, is necessary for vestibular function and survival in zebrafish. J Neurobiol 43:329–337

    Article  CAS  PubMed  Google Scholar 

  • Rubel EW, Fritzsch B (2002) Auditory system development: primary auditory neurons and their targets. Annu Rev Neurosci 25:51–101

    Article  CAS  PubMed  Google Scholar 

  • Schraders M et al (2012) Mutations of the gene encoding otogelin are a cause of autosomal-recessive nonsyndromic moderate hearing impairment. Am J Hum Genet 91:883–889

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sebastian C, Esseling K, Horn E (2001) Altered gravitational forces affect the development of the static vestibuloocular reflex in fish (Oreochromis mossambicus). J Neurobiol 46:59–72

    Article  CAS  PubMed  Google Scholar 

  • Simmler MC, Cohen-Salmon M, El-Amraoui A, Guillaud L, Benichou JC, Petit C, Panthier JJ (2000a) Targeted disruption of otog results in deafness and severe imbalance. Nat Genet 24:139–143

    Article  CAS  PubMed  Google Scholar 

  • Simmler MC, Zwaenepoel II, Verpy E, Guillaud L, Elbaz C, Petit C, Panthier JJ (2000b) Twister mutant mice are defective for otogelin, a component specific to inner ear acellular membranes. Mamm Genome 11:961–966

    Article  CAS  PubMed  Google Scholar 

  • Stooke-Vaughan GA, Obholzer ND, Baxendale S, Megason SG, Whitfield TT (2015) Otolith tethering in the zebrafish otic vesicle requires Otogelin and alpha-Tectorin. Development 142:1137–1145

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Straka H (2010) Ontogenetic rules and constraints of vestibulo-ocular reflex development. Curr Opin Neurobiol 20:689–695

    Article  CAS  PubMed  Google Scholar 

  • Straka H, Fritzsch B, Glover JC (2014) Connecting ears to eye muscles: evolution of a ‘simple’ reflex arc. Brain Behav Evol 83:162–175

    Article  PubMed  Google Scholar 

  • Torborg CL, Feller MB (2005) Spontaneous patterned retinal activity and the refinement of retinal projections. Prog Neurobiol 76:213–235

    Article  PubMed  Google Scholar 

  • Whitfield TT, Granato M, van Eeden FJ, Schach U, Brand M, Furutani-Seiki M, Haffter P, Hammerschmidt M, Heisenberg CP, Jiang YJ, Kane DA, Kelsh RN, Mullins MC, Odenthal J, Nusslein-Volhard C (1996) Mutations affecting development of the zebrafish inner ear and lateral line. Development 123:241–254

    CAS  PubMed  Google Scholar 

  • Wolman MA, Jain RA, Liss L, Granato M (2011) Chemical modulation of memory formation in larval zebrafish. Proc Natl Acad Sci U S A 108:15468–15473

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu LQ, Dickman JD (2012) Neural correlates of a magnetic sense. Science 336:1054–1057

    Article  CAS  PubMed  Google Scholar 

  • Yariz KO et al (2012) Mutations in OTOGL, encoding the inner ear protein otogelin-like, cause moderate sensorineural hearing loss. Am J Hum Genet 91:872–882

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors are grateful to Dr. Teresa Nicolson for sharing the rock solo line, Dr. Mark Warchol for the otoferlin antibody and protocol, and to Dr. Rebecca Callahan for critical feedback on the manuscript. Funding for this research was provided by the National Institute for Deafness and Other Communication Disorders (R00DC012536); by the Pew Scholars Program; and by an Alfred P. Sloan Fellowship to M.W.B.

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Authors and Affiliations

  1. Department of Neuroscience, Washington University School of Medicine, 660 S. Euclid Ave, CB 8108, St. Louis, MO, 63110, USA

    Richard Roberts, Jeffrey Elsner & Martha W. Bagnall

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  1. Richard Roberts
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  2. Jeffrey Elsner
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  3. Martha W. Bagnall
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Correspondence to Martha W. Bagnall.

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Richard Roberts and Jeffrey Elsner contributed equally to the manuscript.

Electronic supplementary material

Video 1

(Rocksolo_14dpf_abnormalswim.avi) presents a high-speed video (500 frames/s) of a otog c.1522+2T>A animal swimming abnormally on the first day of utricular otolith appearance. 45 ° mirrors to reflect a side view can be seen in the upper and lower left of the image. The animal is initially side-lying; after being touched gently with a plastic probe, it turns and swims entirely upside-down (note the ventral swim bladder can be seen on the upper surface in the mirror views) before eventually rolling back to side-lying. The entire video is 480 ms long. (M4V 1531 kb)

Video 2

(Rocksolo_15dpf_normalswim.avi) presents the same animal as in Video 1, 1 day later. Three swim episodes are shown with brief periods of quiescence in between removed. The animal swims normally (dorsal up) for both spontaneous and light touch-evoked swims. The swim bladder can be seen in side mirror view clearly on the underside of the animal. The video is 746 ms long. (M4V 2568 kb)

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Roberts, R., Elsner, J. & Bagnall, M.W. Delayed Otolith Development Does Not Impair Vestibular Circuit Formation in Zebrafish. JARO 18, 415–425 (2017). https://doi.org/10.1007/s10162-017-0617-9

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  • DOI: https://doi.org/10.1007/s10162-017-0617-9

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