Early Hearing Loss upon Disruption of Slc4a10 in C57BL/6 Mice

  • Antje K. Huebner
  • Hannes Maier
  • Alena Maul
  • Sandor Nietzsche
  • Tanja Herrmann
  • Jeppe Praetorius
  • Christian A. HübnerEmail author
Research Article


The unique composition of the endolymph with a high extracellular K+ concentration is essential for sensory transduction in the inner ear. It is secreted by a specialized epithelium, the stria vascularis, that is connected to the fibrocyte meshwork of the spiral ligament in the lateral wall of the cochlea via gap junctions. In this study, we show that in mice the expression of the bicarbonate transporter Slc4a10/Ncbe/Nbcn2 in spiral ligament fibrocytes starts shortly before hearing onset. Its disruption in a C57BL/6 background results in early onset progressive hearing loss. This hearing loss is characterized by a reduced endocochlear potential from hearing onset onward and progressive degeneration of outer hair cells. Notably, the expression of a related bicarbonate transporter, i.e., Slc4a7/Nbcn1, is also lost in spiral ligament fibrocytes of Slc4a10 knockout mice. The histological analysis of the spiral ligament of Slc4a10 knockout mice does not reveal overt fibrocyte loss as reported for Slc4a7 knockout mice. The ultrastructural analysis, however, shows mitochondrial alterations in fibrocytes of Slc4a10 knockout mice. Our data suggest that Slc4a10 and Slc4a7 are functionally related and essential for inner ear homeostasis.


bicarbonate transport Slc4a10 Slc4a7 NCBE deafness fibrocyte pH 



We thank Cynthia Halvorson for the final editing of the manuscript.

Author Contributions

H.M, A.M., S.N., T.H.: performed experiments, analyzed data, and wrote the paper.

A.K..: performed experiments, analyzed data, and wrote the paper.

J.P.: provided material and wrote the paper.

C.A.H.: initiated the study, supervised experiments, and wrote the paper.


This study was funded by grants of the DFG to C.A.H. (HU 800/5-1 and HU 800/8-1).

Compliance with Ethical Standards

All experiments were approved by the responsible local institution (Landesamt für Lebensmittelsicherheit und Verbraucherschutz, Bad Langensalza and Niedersächsisches Landesamt für Verbraucherschutz und Lebensmittelsicherheit, Germany) and comply with the ARRIVE guidelines.

Competing Interests

The authors declare that they have no conflict of interest.


  1. Boedtkjer E, Praetorius J, Matchkov VV, Stankevicius E, Mogensen S, Fuchtbauer AC, Simonsen U, Fuchtbauer EM, Aalkjaer C (2011) Disruption of Na+,HCO3 cotransporter NBCn1 (slc4a7) inhibits NO-mediated vasorelaxation, smooth muscle Ca2+ sensitivity, and hypertension development in mice. Circulation 124:1819–1829CrossRefPubMedGoogle Scholar
  2. Boettger T, Hubner CA, Maier H, Rust MB, Beck FX, Jentsch TJ (2002) Deafness and renal tubular acidosis in mice lacking the K-Cl co-transporter Kcc4. Nature 416:874–878CrossRefPubMedGoogle Scholar
  3. Boettger T, Rust MB, Maier H, Seidenbecher T, Schweizer M, Keating DJ, Faulhaber J, Ehmke H, Pfeffer C, Scheel O, Lemcke B, Horst J, Leuwer R, Pape HC, Volkl H, Hubner CA, Jentsch TJ (2003) Loss of K-Cl co-transporter KCC3 causes deafness, neurodegeneration and reduced seizure threshold. EMBO J 22:5422–5434CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bok D, Galbraith G, Lopez I, Woodruff M, Nusinowitz S, BeltrandelRio H, Huang W, Zhao S, Geske R, Montgomery C, Van Sligtenhorst I, Friddle C, Platt K, Sparks MJ, Pushkin A, Abuladze N, Ishiyama A, Dukkipati R, Liu W, Kurtz I (2003) Blindness and auditory impairment caused by loss of the sodium bicarbonate cotransporter NBC3. Nat Genet 34:313–319CrossRefPubMedGoogle Scholar
  5. Cohen-Salmon M, Ott T, Michel V, Hardelin JP, Perfettini I, Eybalin M, Wu T, Marcus DC, Wangemann P, Willecke K, Petit C (2002) Targeted ablation of connexin26 in the inner ear epithelial gap junction network causes hearing impairment and cell death. Curr Biol 12:1106–1111CrossRefPubMedPubMedCentralGoogle Scholar
  6. Desir J, Moya G, Reish O, Van Regemorter N, Deconinck H, David KL, Meire FM, Abramowicz MJ (2007) Borate transporter SLC4A11 mutations cause both Harboyan syndrome and non-syndromic corneal endothelial dystrophy. J Med Genet 44:322–326CrossRefPubMedPubMedCentralGoogle Scholar
  7. Everett LA, Belyantseva IA, Noben-Trauth K, Cantos R, Chen A, Thakkar SI, Hoogstraten-Miller SL, Kachar B, Wu DK, Green ED (2001) Targeted disruption of mouse Pds provides insight about the inner-ear defects encountered in Pendred syndrome. Hum Mol Genet 10:153–161CrossRefPubMedGoogle Scholar
  8. Francis D, Stergiopoulos K, Ek-Vitorin JF, Cao FL, Taffet SM, Delmar M (1999) Connexin diversity and gap junction regulation by pHi. Dev Genet 24:123–136CrossRefPubMedGoogle Scholar
  9. Groger N, Frohlich H, Maier H, Olbrich A, Kostin S, Braun T, Boettger T (2010) SLC4A11 prevents osmotic imbalance leading to corneal endothelial dystrophy, deafness, and polyuria. J Biol Chem 285:14467–14474CrossRefPubMedPubMedCentralGoogle Scholar
  10. Han C, Someya S (2013) Mouse models of age-related mitochondrial neurosensory hearing loss. Mol Cell Neurosci 55:95–100CrossRefPubMedGoogle Scholar
  11. Hennings JC, Picard N, Huebner AK, Stauber T, Maier H, Brown D, Jentsch TJ, Vargas-Poussou R, Eladari D, Hubner CA (2012) A mouse model for distal renal tubular acidosis reveals a previously unrecognized role of the V-ATPase a4 subunit in the proximal tubule. EMBO mol med 4:1057–1071Google Scholar
  12. Henry KR, Chole RA (1980) Genotypic differences in behavioral, physiological and anatomical expressions of age-related hearing loss in the laboratory mouse. Audiology 19:369–383CrossRefPubMedGoogle Scholar
  13. Hilgen G, Huebner AK, Tanimoto N, Sothilingam V, Seide C, Garrido MG, Schmidt KF, Seeliger MW, Lowel S, Weiler R, Hubner CA, Dedek K (2012) Lack of the sodium-driven chloride bicarbonate exchanger NCBE impairs visual function in the mouse retina. PLoS One 7:e46155CrossRefPubMedPubMedCentralGoogle Scholar
  14. Jacobs S, Ruusuvuori E, Sipila ST, Haapanen A, Damkier HH, Kurth I, Hentschke M, Schweizer M, Rudhard Y, Laatikainen LM, Tyynela J, Praetorius J, Voipio J, Hubner CA (2008) Mice with targeted Slc4a10 gene disruption have small brain ventricles and show reduced neuronal excitability. Proc Natl Acad Sci U S A 105:311–316CrossRefPubMedGoogle Scholar
  15. Johnson KR, Zheng QY, Noben-Trauth K (2006) Strain background effects and genetic modifiers of hearing in mice. Brain Res 1091:79–88CrossRefPubMedPubMedCentralGoogle Scholar
  16. Lang F, Vallon V, Knipper M, Wangemann P (2007) Functional significance of channels and transporters expressed in the inner ear and kidney. Am J Physiol 293:C1187–C1208CrossRefGoogle Scholar
  17. Le Calvez S, Avan P, Gilain L, Romand R (1998) CD1 hearing-impaired mice. I: distortion product otoacoustic emission levels, cochlear function and morphology. Hear Res 120:37–50CrossRefPubMedGoogle Scholar
  18. Lopez IA, Acuna D, Galbraith G, Bok D, Ishiyama A, Liu W, Kurtz I (2005) Time course of auditory impairment in mice lacking the electroneutral sodium bicarbonate cotransporter NBC3 (slc4a7). Brain Res 160:63–77CrossRefGoogle Scholar
  19. Lopez-Bigas N, Arbones ML, Estivill X, Simonneau L (2002) Expression profiles of the connexin genes, Gjb1 and Gjb3, in the developing mouse cochlea. Mech Dev 119(Suppl 1):S111–S115CrossRefPubMedGoogle Scholar
  20. Mahendrasingam S, Macdonald JA, Furness DN (2011) Relative time course of degeneration of different cochlear structures in the CD/1 mouse model of accelerated aging. JARO 12:437–453CrossRefPubMedGoogle Scholar
  21. Markel P, Shu P, Ebeling C, Carlson GA, Nagle DL, Smutko JS, Moore KJ (1997) Theoretical and empirical issues for marker-assisted breeding of congenic mouse strains. Nat Genet 17:280–284CrossRefPubMedGoogle Scholar
  22. Palacios-Prado N, Briggs SW, Skeberdis VA, Pranevicius M, Bennett MV, Bukauskas FF (2010) pH-dependent modulation of voltage gating in connexin45 homotypic and connexin45/connexin43 heterotypic gap junctions. Proc Natl Acad Sci U S A 107:9897–9902CrossRefPubMedPubMedCentralGoogle Scholar
  23. Park M, Li Q, Shcheynikov N, Zeng W, Muallem S (2004) NaBC1 is a ubiquitous electrogenic Na+-coupled borate transporter essential for cellular boron homeostasis and cell growth and proliferation. Mol Cell 16:331–341CrossRefPubMedGoogle Scholar
  24. Parker M, Bitner-Glindzicz M (2015) Genetic investigations in childhood deafness. Arch Dis Child 100:271–278CrossRefPubMedGoogle Scholar
  25. Parker MD, Musa-Aziz R, Rojas JD, Choi I, Daly CM, Boron WF (2008) Characterization of human SLC4A10 as an electroneutral Na/HCO3 cotransporter (NBCn2) with Cl- self-exchange activity. J Biol Chem 283:12777–12788CrossRefPubMedPubMedCentralGoogle Scholar
  26. Petit C (2006) From deafness genes to hearing mechanisms: harmony and counterpoint. Trends Mol Med 12:57–64CrossRefPubMedGoogle Scholar
  27. Petit C, Levilliers J, Hardelin JP (2001) Molecular genetics of hearing loss. Annu Rev Genet 35:589–646CrossRefPubMedGoogle Scholar
  28. Potter PK, Bowl MR, Jeyarajan P, Wisby L, Blease A, Goldsworthy ME, Simon MM, Greenaway S, Michel V, Barnard A, Aguilar C, Agnew T, Banks G, Blake A, Chessum L, Dorning J, Falcone S, Goosey L, Harris S, Haynes A, Heise I, Hillier R, Hough T, Hoslin A, Hutchison M, King R, Kumar S, Lad HV, Law G, MacLaren RE, Morse S, Nicol T, Parker A, Pickford K, Sethi S, Starbuck B, Stelma F, Cheeseman M, Cross SH, Foster RG, Jackson IJ, Peirson SN, Thakker RV, Vincent T, Scudamore C, Wells S, el-Amraoui A, Petit C, Acevedo-Arozena A, Nolan PM, Cox R, Mallon AM, Brown SDM (2016) Novel gene function revealed by mouse mutagenesis screens for models of age-related disease. Nat Commun 7:12444CrossRefPubMedPubMedCentralGoogle Scholar
  29. Romero MF, Chen AP, Parker MD, Boron WF (2013) The SLC4 family of bicarbonate (HCO3 ) transporters. Mol Asp Med 34:159–182CrossRefGoogle Scholar
  30. Sadanaga M, Morimitsu T (1995) Development of endocochlear potential and its negative component in mouse cochlea. Hear Res 89:155–161CrossRefPubMedGoogle Scholar
  31. Sinning A, Liebmann L, Hubner CA (2015) Disruption of Slc4a10 augments neuronal excitability and modulates synaptic short-term plasticity. Front Cell Neurosci 9:223CrossRefPubMedPubMedCentralGoogle Scholar
  32. Spicer SS, Schulte BA (1991) Differentiation of inner ear fibrocytes according to their ion transport related activity. Hear Res 56:53–64CrossRefPubMedGoogle Scholar
  33. Spray DC, Harris AL, Bennett MV (1981) Gap junctional conductance is a simple and sensitive function of intracellular pH. Science 211:712–715CrossRefPubMedGoogle Scholar
  34. Stergiopoulos K, Alvarado JL, Mastroianni M, Ek-Vitorin JF, Taffet SM, Delmar M (1999) Hetero-domain interactions as a mechanism for the regulation of connexin channels. Circ Res 84:1144–1155CrossRefPubMedGoogle Scholar
  35. Teubner B, Michel V, Pesch J, Lautermann J, Cohen-Salmon M, Sohl G, Jahnke K, Winterhager E, Herberhold C, Hardelin JP, Petit C, Willecke K (2003) Connexin30 (Gjb6)-deficiency causes severe hearing impairment and lack of endocochlear potential. Hum Mol Genet 12:13–21CrossRefPubMedGoogle Scholar
  36. Wang CZ, Yano H, Nagashima K, Seino S (2000) The Na+-driven Cl/HCO3 exchanger. Cloning, tissue distribution, and functional characterization. J Biol Chem 275:35486–35490CrossRefPubMedGoogle Scholar
  37. Wangemann P, Itza EM, Albrecht B, Wu T, Jabba SV, Maganti RJ, Lee JH, Everett LA, Wall SM, Royaux IE, Green ED, Marcus DC (2004) Loss of KCNJ10 protein expression abolishes endocochlear potential and causes deafness in Pendred syndrome mouse model. BMC Med 2:30CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Association for Research in Otolaryngology 2019

Authors and Affiliations

  • Antje K. Huebner
    • 1
  • Hannes Maier
    • 2
  • Alena Maul
    • 3
  • Sandor Nietzsche
    • 4
  • Tanja Herrmann
    • 1
  • Jeppe Praetorius
    • 5
  • Christian A. Hübner
    • 1
    Email author
  1. 1.Institute of Human GeneticsJena University Hospital, Friedrich Schiller UniversitätJenaGermany
  2. 2.Department of Otolaryngology and Cluster of Excellence Hearing4all, Deutsches HörZentrum HannoverMedical University HannoverHannoverGermany
  3. 3.Max-Delbrück Centrum für Molekulare Medizin (MDC) and NeuroCureBerlinGermany
  4. 4.Electron Microscopy CenterJena University Hospital, Friedrich Schiller UniversitätJenaGermany
  5. 5.Department of Biomedicine, HealthAarhus UniversityAarhusDenmark

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